detecção de citocinas em culturas de linfócitos em pacientes
TRANSCRIPT
LUIS EDUARDO AGNER MACHADO MARTINS
'HWHFomR�GH�FLWRFLQDV�HP�FXOWXUDV�GH�OLQIyFLWRV�HP�
SDFLHQWHV�DOpUJLFRV�DR�FURPR
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GD�8QLYHUVLGDGH�GH�6mR�3DXOR�SDUD�REWHQomR�
GR�WtWXOR�GH�Doutor em Dematologia
Programa: Dermatologia
Orientador: Dr. Vitor Manoel Silva dos Reis
6mR�3DXOR
2010
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Dados Internacionais de Catalogação na Publicação (CIP)
Preparada pela Biblioteca da Faculdade de Medicina da Universidade de São Paulo
©reprodução autorizada pelo autor
Martins, Luis Eduardo Agner Machado Detecção de citocinas em culturas de linfócitos em pacientes alérgicos ao cromo / Luis Eduardo Agner Machado Martins. -- São Paulo, 2010.
Tese(doutorado)--Faculdade de Medicina da Universidade de São Paulo. Programa de Dermatologia.
Orientador: Vitor Manoel Silva dos Reis.
Descritores: 1.Teste de proliferação linfocitária 2.Hipersensibilidade/diagnóstico 3.Cromo 4.Citocinas 5.Dermatite de contato
USP/FM/DBD-229/10
DEDICATÏRIA
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Aos meus pais pelo constante
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AGRADECIMENTOS
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Ao Profo. Dr. Vitor Manoel Silva dos Reis que mais uma vez acreditou no meu
potencial e contribuiu ativamente durante todo o desenvolvimento desse
projeto.
Ao Profo. Dr. Evandro Rivitti e PHPEURV� GD� FRPLVVmR� GD� SyV-JUDGXDomR� GR�
Departamento de Dermatologia que permitiram a continuidade do trabalho.
Ao Profo�'U��$OEHUWR�-RVp�GD�6LOYD�'XDUWH, a Profa. Dra. Mirian Nacagami Sotto e
demais integrantes do LIM-���SRU�DSRLDU�H�IRUQHFHU�RV�PHLRV�SDUD�D�UHDOL]DomR�
da tese.
A Profa. Dra. 9DOpULD�$RNL�e ao Profo. Dr. Paulo Ricardo Criado pela amizade,
DX[tOLR�H�RULHQWDo}HV�SUHVWDGDV�
$R�5LFDUGR�6SLQD�1XQHV�SHOD�GHGLFDomR�H�UHDOL]DomR�GRV�WHVWHV�GH�FRQWDWR�
A todos os demais professores, residentes� H� IXQFLRQiULRV� pela amizade e
carinho.
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Lista de abreviaturas/LVWD�GH�VtPERORVLista de tabelas�H�JUiILFRV ResumoSummary ��,1752'8d2.......................................................................................... 012 OBJETIVOS................................................................................................ 06��5(9,62�'$�/,7(5$785$................................................................... 073.1.1 Cromo................................................................................................. 07 3.1.2 0DQLIHVWDo}HV�FOtQLFDV�GD�'&$�SHOR�FURPR....................................... 08 3.1.3 3URJQyVWLFR�GD�'&$�SRU�FURPR.......................................................... 09 3.2 3DWRJrQHVH�GD�GHUPDWLWH�GH�FRQWDWR�DOpUJLFD........................................ 10 3.2.1 Fase aferente ..................................................................................... 12 3.2.2 Fase eferente ,.................................................................................... 153.2.3 &pOXODV�HQYROYLGDV.............................................................................. 16 3.2.3.��&pOXODV�GHQGUtWLFDV........................................................................... 163.2.3.1.1 As &pOXODV�GHQGUtWLFDV�QD�IDVH�DIHUHQWH........................................ 173.2.3.1.2 As FpOXODV�GHQGUtWLFDV�QD�IDVH�HIHUHQWH......................................... 20 3.2.3.2 /LQIyFLWRV ........................................................................................ 213.2.3.2.1 /LQIyFLWRV�HIHWRUHV......................................................................... 21����������/LQIyFLWRV�7�UHJXODGRUHV����������...................................................... 23��������4XHUDWLQyFLWRV.................................................................................. 263.2.3.4 0DVWyFLWRV........................................................................................ 283.2.3.5 /LQIyFLWRV�%�H�FpOulas T NK.............................................................. 28 4 7HVWHV�GLDJQyVWLFRV.................................................................................. 304.1 Teste de contato ................................................................................... 304.2 7HVWH�GH�SUROLIHUDomR�liQIRFLWiULD............................................................ 31 ��0e72'26............................................................................................... 42����6HOHomR�GH�SDFLHQWHV�������������������������������������������������������..................... 424.2 Teste de contato.................................................................................... 43 ����7HVWH�GH�SUROLIHUDomR�OLQIRFLWiULD���������������������������������������..................... 454.3.1 7HVWH�GH�SUROLIHUDomR�OLQIRFLWiULD�SDUD�GHWHFomR�GD�DWLYDomR�FHOXODU�pela taxa GH�FDSWDomR�GH�WLPLGLQD�UDGLRPDUFDGD........................................ 47 4.3.2 7HVWH�GH�SUROLIHUDomR�OLQIRFLWiULD�SDUD�GHWHFomR�GD�DWLYDomR�FHOXODUpela pURGXomR de citocinas ......................................................................... 49 4.4 Dosagem das citocinas no sobrenadante das culturas ........................ 50 4.4.1 ³/XPLQH[�OLTXLFKLS�DUUD\´................................................................... 50 4.4.2 Citocinas testadas ............................................................................ 52 ����0pWRGRV�HVWDWtVWLFRV�����������������������������������������������������������.................. 535 RESULTADOS........................................................................................ 545.1 6HOHomR�GRV�JUXSRV.............................................................................. 545.2 Testes de contato................................................................................. 54
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5.3�7HVWHV�GH�SUROLIHUDomR�OLQIRFLWiULD������������������������������������������������......... 55 5.4 'HWHFomR�GDV�FLWRFLQDV�........................................................................ 575.4.1 Interferon-gama............................................................................... 575.4.2 Interlucina-2..................................................................................... 59 5.4.3 Interlucina-4..................................................................................... 61 5.4.4 Interlucina-5..................................................................................... 63 5.4.5 Interlucina-10................................................................................... 65 5.4.6 Interlucina-12................................................................................... 67 5.4.7 Interlucina-13................................................................................... 69 5.4.8 Interlucina-17................................................................................... 71 5.4.9 RANTES.......................................................................................... 73����7HVWHV�GH�FRSUUHODomR.......................................................................... 75��',6&8662........................................................................................... 767 &21&/86®(6....................................................................................... 868 ANEXOS................................................................................................. 87 ��5()(5Ç1&,$6�%,%/,2*5È),&$6...................................................... 100
LISTAS
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Lista de Abreviaturas
Ag = $QWtJHQR C5 = )UDomR�&��GR�FRPSOHPHQWR C5a = )UDomR�&�D�GR�FRPSOHPHQWR CCL1 = Quimiocina CCL1 CCL5 = Quimiocina CCL5 (RANTES)
CCL19 = Quimiocina CCL19 CCL21 = Quimiocina CCL21 CCR4 = Receptor de quimiocina CCR4 CCR7 = Receptor de quimiocina CCR7 CCR8 = Receptor de quimiocina CCR8 CD = &pOXOD�GHQWUtWLFD CD1d = 0HPEUR�GD�IDPtOLD�&'� CD4+ = $QWtJHQR�&'� CD4+CD25+ = &pOXOD�7�UHJXODGRUD�&'�+CD25+
CD8+ = $QWtJHQR�&'� CD25 = $QWtJHQR�&'�� CD28 = $QWtJHQR�&'�� CD80 = $QWtJHQR�&'�� CD86 = $QWtJHQR�&'�� CD205 = $QWtJHQR�&'��� CLA = $QWtJHQR�OLQIRFLWiULR�FXWkQHR CL = &pOXOD�GH�/DQJHUKDQV
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Cpm = Contagens por minuto
Cr (III) = Cromo trivalente Cr (VI) = Cromo hexavalente CrCl3 = Cloreto de cromo CTLA-4 = $QWtJHQR���DVVRFLDGR�DR�OLQIyFLWR�7�FLWRWy[LFR DC = Dermatite de contato
DCA = 'HUPDWLWH�GH�FRQWDWR�DOpUJLFD DCI = Dermatite DNA = ÈFLGR�GHVR[LUULERQXFOpLFR DTH = 5HDomR�GH�KLSHUVHQVLELOLGDGH�WDUGLD ELISA = Ensaio imunoenzLPiWLFR ELISpot = EQVDLR�LPXQRHQ]LPiWLFR modificado Fas = 3URWHtQD�)DV FasL = /LJDQWH�GD�SURWHtQD�)DV Fas-FasL = ,QWHUDomR�GD�SURWHtQD�)DV�FRP�VHX�OLJDQWH Foxp3 = )DWRU�GH�WUDQVFULomR�QXFOHDU�)R[S� GM-CSF = Fator estimuladRU�GD�FRO{QLD�GH�PDFUyIDJRV�H�PRQyFLWRV� HC-FMUSP = +RVSLWDO�GH�&OtQLFDV�GD�)DFXOGDGH�GH�0HGLFLQD�GD�
8QLYHUVLGDGH�6mR�3DXOR� ICAM-1 = 0ROpFXOD�GH�DGHVmR�LQWHUFHOXODU IE = ËQGLFH�GH�HVWLPXODomR IgM = Imunoglobulina M IL-1 = Interleucina 1
IL-1 = Interleucina 1 alfa IL-1 = Interleucina 1 beta IL-2 = Interleucina 2 IL-4 = Interleucina 4 IL-5 = Interleucina 5 IL-6 = Interleucina 6
IL-7 = Interleucina 7 IL-8 = Interleucina 8 IL-10 = Interleucina 10 IL-12 = Interleucina 12 IL-13 = Interleucina 13 IL-16 = Interleucina 16 IL-17 = Interleucina 17 IL-23 = Interleucina 23 INF- = Interferon gama K2Cr2O7 = %LFURPDWR�GH�SRWiVVLR LB = /LQIyFLWR�% LFA-1 = 3URWHtQD���DVVRFLDGD�D�IXQomR�OLQIRFLWiULD LT = /LQIyFLWR�7 LT NK = FpOXODV�7�H[WHUPLQDGRUDV�QDWXUDLV M = Molar MHC I = Complexo principal de histocompatibilidade 1 MHC II = Complexo principal de histocompatibilidade 2
NK = &pOXOD�H[WHUPLQDGRUD�QDWXUDl P = 6LJQLILFkQFLD PWM = Pokeweed RANK = Receptor ativador do fator nuclear B RANKL = Ligante do receptor ativador do fator nuclear B RANTES = Quimiocina RANTES (CCL5) RNAm = ÈFLGR�ULERQXFOpLFR�PHQVVDJHLUR Rpm = 5RWDo}HV�SRU�PLQXWR RPMI 1640 = Meio de cultura Rosewell Park Memorial Institute TC = Teste de contato Tc1 = 7�FLWRWy[LFD�WLSR�� TGF- = Fator transformador do crescimento beta Th1 = T auxiliar tipo 1 Th2 = T auxiliar tipo 2 Th17 = T auxiliar tipo 17 TNF- = Fator de necrose tumoral alfa TPL = 7HVWH�GH�SUROLIHUDomR�OLQIRFLWiULD Tr1 = /LQIyFLWR�7�UHJXODGRU�� UVB = 5DGLDomR�XOWUDYLROHWD�% VCAM-1 = 0ROpFXOD�1 GH�DGHVmR�GD�FpOXOD�YDVFXODU VLA-4 = $QWtJHQR�GH�DWLYDomR�PXLWR�WDUGLD�� VLA-5 = $QWtJHQR�GH�DWLYDomR�PXLWR�WDUGLD��
VLA-6 = $QWtJHQR�GH�DWLYDomR�PXLWR�WDUGLD��
/LVWD�GH�6tPERORV
Delta
g Micrograma
g Nanograma
L Microlitro M Molar > Maior que
Maior ou igual a < Menor que % por cento
oC grau Celsius
/LVWD�GH�7DEHODV�H�*UiILFRV
FIGURA 1 0HFDQLVPR�GD�VHQVLELOL]DomR�H�HOLFLWDomR�GD�GHUPDWLWH�GH�FRQWDWR�
DOpUJLFD..........................................................................................11 FIGURA 2 Microesferas de poliestireno coradas de forma singular...............50 FIGURA 3 $�HVIHUD�FRP�R�DQWLFRUSR�p�LQFXEDGD�FRP�D�DPRVWUD�H�VH�OLJD�DR�
VHX�DQWtJHQR��$� DPRVWUD� p� ODYDGD�H� LQFXEDGD� FRP�XP�DQWLFRUSR�biotinilado que se ligD� DR� FRPSOH[R� DQWtJHQR-anticorpo. O resultado p amplificado pela� LQFXEDomR� FRP� R� FRQMXJDGR�estreptavidina-ficoeritrina que emite um sinal fluorescente ..........51
FIGURA 4 2V�IHL[HV�GH�ODVHU�LQFLGHP�VREUH�DV�HVIHUDV�SHUPLWLQGR�D�GHWHFomR�
do ensaio e da intensidade da resposta........................................52 FIGURA 5 Contagens por minuto encontradas nas culturas basais e
estimuladas com PWM do grupo de casos. .................................56 FIGURA 6 Contagens por minuto encontradas nas culturas basais e
estimuladas com PWM do grupo controle.....................................56 FIGURA 7 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QRV� FDVRV� H� FRQWUROHV�
estimulados com PWM. ................................................................56 FIGURA 8 Contagens por minuto encontradas nas culturas basais e
estimuladas com cromo do grupo de casos. ................................56 FIGURA 9 Contagens por minuto encontradas nas culturas basais e
estimuladas com cromo do grupo controle. ..................................56 FIGURA 10 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QRV� FDVRV� H� FRQWUROHV�
estimulados com cromo.................................................................56 FIGURA 11 'HWHFomR�GH�,)1- nas culturas basais dos casos e controles.....58 FIGURA 12 'HWHFomR�GH� ,)1- nas culturas basais e estimuladas com cromo
do grupo de casos.........................................................................58 FIGURA 13 'HWHFomR�GH� ,)1- nas culturas basais e estimuladas com cromo
do grupo controle. .........................................................................58
FIGURA 14 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDVRV� H�controles........................................................................................58
FIGURA 15 'LIHUHQoD� QD� GHWHFomR� GH� ,)1- entre as culturas estimuladas e basais dos grupos de caso e controle...........................................58
FIGURA 16 'HWHFomR�GH�,/-2 nas culturas basais dos casos e controles........60 FIGURA 17 'HWHFomR�GH�,/-2 nas culturas basais e estimuladas com cromo do
grupo de casos. ............................................................................60 FIGURA 18 'HWHFomR�GH�,/-2 nas culturas basais e estimuladas com cromo do
grupo controle. ..............................................................................60 FIGURA 19 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDVRV� H
controles........................................................................................60 FIGURA 20 'LIHUHQoD�QD�GHWHFomR�GH�,/-2 entre as culturas de estimuladas e
basais dos grupos de caso e controle. .........................................60 FIGURA 21 'HWHFomR�GH�,/-4 nas culturas basais dos casos e controles........62 FIGURA 22 'HWHFomR�GH�,/-4 nas culturas basais e estimuladas com cromo do
grupo de casos..............................................................................62 FIGURA 23 'HWHFomR�GH�,/-4 nas culturas basais e estimuladas com cromo do
grupo controle................................................................................62 FIGURA 24 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV culturas dos casos e
controles........................................................................................62 FIGURA 25 'LIHUHQoD� QD� GHWHFomR� GH� ,/-4 entre as culturas estimuladas e
basais dos grupos de caso e controle. .........................................62 FIGURA 26 'HWHFomR�GH�,/-5 nas culturas basais dos casos e controles........64 FIGURA 27 'HWHFomR�GH�,/-5 nas culturas basais e estimuladas com cromo do
grupo de casos. ............................................................................64 FIGURA 28 'HWHFomR�GH�,/-5 nas culturas basais e estimuladas com cromo do
grupo controle................................................................................64 FIGURA 29 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDVRV� H�
controles........................................................................................64
FIGURA 30 'LIHUHQoD� QD� GHWHFomR� GH� ,/-5 entre as culturas estimuladas e basais dos grupos de caso e controle. .........................................64
FIGURA 31 DeteFomR�GH�,/-10 nas culturas basais dos casos e controles......66 FIGURA 32 'HWHFomR�GH� ,/-10 nas culturas basais e estimuladas com cromo
do grupo de casos. .......................................................................66 FIGURA 33 'HWHFomR�GH� ,/-10 nas culturas basais e estimuladas com cromo
do grupo controle...........................................................................66 FIGURA 34 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDVRV� H�
controles........................................................................................66 FIGURA 35 'LIHUHQoD� QD� GHWHFomR� GH� ,/-10 entre as culturas estimuladas e
basais dos grupos de caso e controle...........................................66 FIGURA 36 'HWHFomR�GH�,/-12 nas culturas basais dos casos e controles......68 FIGURA 37 'HWHFomR�GH� ,/-12 nas culturas basais e estimuladas com cromo
do grupo de casos.........................................................................68 FIGURA 38 'HWHFomR�GH� ,/-12 nas culturas basais e estimuladas com cromo
do grupo controle...........................................................................68 FIGURA 39 ËQGLFHV� GH� HVWLPXODomR� HQFontrados nas culturas dos casos e
controles........................................................................................68 FIGURA 40 'LIHUHQoD� QD� GHWHFomR� GH� ,/-12 entre as culturas estimuladas e
basais dos grupos de caso e controle...........................................68 FIGURA 41 'HWHFomR�GH�,/-13 nas culturas basais dos casos e controles......70 FIGURA 42 'HWHFomR�GH� ,/-13 nas culturas basais e estimuladas com cromo
do grupo de casos.........................................................................70 FIGURA 43 'HWHFomR�GH� ,/-13 nas culturas basais e estimuladas com cromo
do grupo controle...........................................................................70 FIGURA 44 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDsos e
controles........................................................................................70 FIGURA 45 'LIHUHQoD� QD� GHWHFomR� GH� ,/-13 entre as culturas estimuladas e
basais dos grupos de caso e controle...........................................70 FIGURA 46 'HWHFomR�GH�,/-17 nas culturas basais dos casos e controles......72
FIGURA 47 'HWHFomR�GH� ,/-17 nas culturas basais e estimuladas com cromo
do grupo de casos.........................................................................72 FIGURA 48 'HWHFomR�GH� ,/-17 nas culturas basais e estimuladas com cromo
do grupo controle...........................................................................72 FIGURA 49 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV� GRV� FDVRV� H�
controles........................................................................................72 FIGURA 50 'LIHUHQoD� QD� GHWHFomR� GH� ,/-17 entre as culturas estimuladas e
basais dos grupos de caso e controle. .........................................72
FIGURA 51 'HWHFomR� GH RANTES nas culturas basais dos casos e controles........................................................................................74
FIGURA 52 'HWHFomR� GH� 5$17(6� QDV� FXOWXUDV� EDVDLV� H� HVWLPXODGDV� FRP�
cromo do grupo de casos..............................................................74 FIGURA 53 'HWHFomR� GH� 5$17(6� QDV� FXOWXUDV� EDVDLV� H� HVWLPXODGDV� FRP�
cromo do grupo controle................................................................74 FIGURA 54 ËQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QDV� FXOWXUDV dos casos e
controles........................................................................................74 FIGURA 55 'LIHUHQoD�QD�GHWHFomR�GH�5$17(6�HQWUH�DV�FXOWXUDV�HVWLPXODGDV�
e basais dos grupos de caso e controle. ......................................74 QUADRO 1 7UDEDOKRV�VREUH�WHVWH�GH�SUROLIHUDomR�OLQIRFLWiULD�SRU�FURPR����������� QUADRO 2 *UXSR� GH� VXEVWkQFLDV� WHVWDGDV� QD� EDWHULD� SDGUmR� EUDVLOHLUD� GH�
dermatite de contato......................................................................87 QUADRO 3 *UXSR�GH�VXEVWkQFLDV�WHVWDGDV�QD�EDWHULD�GH�FRVPpWLFRV............87 QUADRO 4 Resultados dos testes de contato.................................................88 TABELA 1 9DORUHV� PpGLRV� GDV� FRQWDJHQV� SRU� PLQXWR� GDV triplicatas dos
casos e controles........................................................................................89
TABELA 2 'HWHFomR� GH� ,)1-gama em pg/mL nas culturas dos casos e
controles........................................................................................90 TABELA 3 'HWHFomR�GH�,/-2 em pg/mL nas culturas dos casos e controles..91
TABELA 4 'HWHFomR�GH�,/-4 em pg/mL nas culturas dos casos e controles..92 TABELA 5 'HWHFomR�GH�,/-5 em pg/mL nas culturas dos casos e controles..93 TABELA 6 'HWHFomR� GH� ,/-10 em pg/mL nas culturas dos casos e
controles........................................................................................94 TABELA 7 'HWHFomR� GH� ,/-12(p70) em pg/mL nas culturas dos casos e
controles........................................................................................95 TABELA 8 'HWHFomR� GH� ,/-13 em pg/mL nas culturas dos casos e
controles........................................................................................96
TABELA 9 'HWHFomR� GH� ,/-17 em pg/mL nas culturas dos casos e controles........................................................................................97
TABELA 10 'HWHFomR� GH� RANTES em pg/mL nas culturas dos casos e
controles........................................................................................98 TABELA 11 &RUUHODomR� HQWUH� DOHUJLD� H� RV� tQGLFHV� GH� HVWLPXODomR� GHWHFWDGRV�
FRP�R�WHVWH�GH�SUROLIHUDomR�OLQIRFLWiULD�.........................................99 TABELA 12 &RUUHODomR�HQWUH�DOHUJLD�H�RV�tQGLFHV�GH�HVWLPXODomR�HQFRQWrados
com as citocinas detectadas. .......................................................99 TABELA 13 &RUUHODomR� HQWUH� Rs tQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� FRP� R�
TPL e RV�tQGLFHV�REWLGRV�FRP�DV�citocinas. ..................................99
RESUMO
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Martins LEAM. 'HWHFomR�GH�FLWRFLQDV�HP�FXOWXUDV�GH�OLQIyFLWRV�HP�SDFLHQWHV�DOpUJLFRV�DR�FURPR. 6mR�3DXOR�� )DFXOGDGH�GH�0HGLFLQD��8QLYHUVLGDGH�6mR�Paulo; 2010.
$� GHUPDWLWH� GH� FRQWDWR� DOpUJLFD� GHFRUUH� GH� XPD� UHDomR� LPXQROyJLFD,PHGLDGD� SRU� FpOXODV� 7�� FRQWra um contactante em pessoas previamente VHQVLELOL]DGDV���2�H[DPH�SDGUmR�RXUR�SDUD�R�GLDJQyVWLFR�GD�'&$�p�R�WHVWH�de contato (TC). Infelizmente, o TC demanda tempo, QmR� p� LQyFXR� e tem DOJXPDV� OLPLWDo}HV� 2� WHVWH�GH�SUROLIHUDomR� OLQIRFLWiULD� �73/��convencional, uma alternativa ao TC, apresenta baixa sensibilidade no GLDJQyVWLFR� GH�alergia ao cromo. A fim de aprimorar a sensibilidade o resultado do teste foi REWLGR� SHOD�GHWHFomR� GH�FLWRFLQDV�QR�VREUHQDGDQWH�GDV�FXOWXUDV�H�QmR�SRU�timidina radiomarcada. DezoitR�SDFLHQWHV�DOpUJLFRV�DR�FURPR�H����FRQWUROHV�foram testados com o teste convencional e com as citocinas IFN- , IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 e rantes. +RXYH� FRUUHODomR� HQWUH� DOHUJLD� DR�cromo e GHWHFomR�GDV�FLWRFLQDV� ,)1- , IL-2, IL-5, IL-12 e IL-13. Dessas, os melhores resultados foram encontrados com a IL-13. O TPL pode ser utilizado como� H[DPH�DGLFLRQDO�RX� DOWHUQDWLYR� QR� GLDJQyVWLFR� GD�'&$�SHOR�cromo.
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SUMMARY
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Martins LEAM. Cytokine detection in lymphocyte cultures of chromium allergic patients [thesis]. )DFXOW\� RI�0HGLFLQH�� 8QLYHUVLW\� RI� 6mR� 3DXOR�� 63�(Brazil); 2010. Allergic contact dermatitis (ACD) elapses from an specific T cell immunologic reaction against an allergen in a sensitized individual. The gold standard exam to confirm ACD is the patch test (PT). However, PT demands time, has potential side efects and some limitations. The standard lymphocyte proliferation assay (LPA) has sensitivity in the diagnosis of chromium allergy. To optimize this test the results were obtained by detection of cytokines instead radiolabeled thymidine. Eighteen patients allergic to chromium and 19 controls were tested with the conventional LPA and for the following cytokines: IFN- , IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 and rantes. Correlation between allergy to chromium and detection of IFN- , IL-2, IL-5, IL-12 e IL-13 was found. The best results were found with IL-13. LPA can be used as an alternative or additional test in the diagnosis of ACD by chromium.
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OBJETIVOS
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3.2 3DWRJrQHVH�GD�Germatite de contato aOpUJLFD
$� '&$� p� XPD� GRHQoD� LQIODPDWyULD� GHVHQFDGHDGD� SRU� KDSWHQRV� H�
PHGLDGD� SRU� FpOXODV� 7�42 Em 1935, Karl Landsteiner e John Jacobs43
HVFUHYHUDP�VREUH�D�H[LVWrQFLD�GH�TXtPLFRV�UHDWLYRV�de baixo peso molecular
TXH� DR� VH� OLJDU� D� SURWHtQDV� GHWHUPLQDP� D� IRUPDomR� GH� DQWLFRUSRV ou
VXEVWkQFLDV� DQWLFRUSR-VtPLOHV, supostamente UHVSRQViYHLV� SHOD� '&$��
6RPHQWH� ��� DQRV� PDLV� WDUGH� 6KHDUHU� GHPRQVWURX� TXH� OLQIyFLWRV� 7 (LT)
hapteno-HVSHFtILFRV� WDPEpP� UHVSRQGHP� D� HVVHV� FRPSOH[RV� KDSWHQR-
proteicos.44
$� '&$� RFRUUH� FRPR� FRQVHT�rQFLD� GH� XPD� cascata de processos
ItVLFR-TXtPLFRV� H� imunes que podem ser didaticamente divididos em duas
IDVHV��GH�LQGXomR��WDPEpP�FKDPDGD�GH�DIHUHQWH�H�GH�HOLFLWDomR�RX�HIHUHQWH�
A figura 1 resume os eventos envolvidos nessas fases.
________________________________________________5HYLVmR�GD�OLWHUDWXUD 11
Fonte: Adaptado de Gober e Gaspari45 e Campos et al.46
Fig 1.� 0HFDQLVPR� GD� VHQVLELOL]DomR� H� HOLFLWDomR� GD� GHUPDWLWH� GH� FRQWDWR�DOpUJLFD�� Na fase aferente os haptenos penetram a pele e se ligam a SURWHtQDV�WHFLGXDLV�WRUQDQGR-VH�DQWtJHQRV��$J��FRPSOHWRV��(VVHV�DQWtJHQRV�VmR�FDSWDGRV� H�SURFHVVDGRV� SHODV�FpOXODV�GHQGUtWLFDV� �&'�� TXH�SDVVDP�D�DSUHVHQWi-ORV� DFRSODGRV� D� PROpFXODV� GR� FRPSOH[R� SULQFLSDO� GH�KLVWRFRPSDWLELOLGDGH� �0+&�� QD� VXSHUItFLH� GD� PHPEUDQD� FHOXODU�� $V� &'�PLJUDP� SDUD� RV� OLQIRQRGRV� UHJLRQDLV� RQGH� DSUHVHQWDP� R� DQWtJHQR� DRV�OLQIyFLWRV� 7� �/7��� 2V� /7� TXH� UHFRQKHFHP� R� DQWtJHQR� DSUHVHQWDGR� VmR�DWLYDGRV��$�SHQHWUDomR�GRV�DQWtJHQRV�QD�SHOH�WDPEpP�GHWHUPLQD�D�OLEHUDomR�GH�JOLFROtSLGHV�HQGyJHQRV�TXH�VmR�DSUHVHQWDGRV�SHODV�&'�SDUD�RV�OLQIyFLWRV�T NK (LT NK). Os LT NK liberam IL-4 que estimula�RV�OLQIyFLWRV�%��/%��WLSR���D� SURGX]LU� ,J0�� )UHQWH� D� XP� QRYR� FRQWDWR� D� LQWHUDomR� GD� ,J0� FRP� R�FRPSOH[R� DQWtJHQR-SURWHLFR� OHYD� D� DWLYDomR� GR� FRPSOHPHQWR� TXH� LQGX]� D�OLEHUDomR� GH� IDWRUHV� TXLPLRWiWLFRV� H� LQIODPDWyULRV� GRV�PDVWyFLWRV� H�FpOXODV�endoteliais. Os LT ativados migram para a pele a interagem com as CD que FDUUHJDP�R�DQWtJHQR�OHYDQGR�D�'&$�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 12
3.2.1 Fase aferente
A fase aferente se desenvolve ao longo do tempo como resultado
GD�H[SRVLomR�UHSHWLGD�D�DJHQWHV�DPELHQWDLV��$�PDLRULD�GRV�FRQWDFWDQWHV�VmR�
JUDQGHV�GHPDLV�SDUD�SDVVDU�R�HVWUDWR�FyUQHR��PDV�RV�KDSWHQRV��GHYLGR�DR�
VHX�EDL[R�SHVR�PROHFXODU�SHQHWUDP�HVVD�FDPDGD�H�VH�GLIXQGHP�HP�GLUHomR�
D�FDPDGD�EDVDO�VHP�VHUHP�UHFRQKHFLGRV�SHOR�VLVWHPD�LPXQROyJLFR��'XUDQWH�
HVVH� SURFHVVR� GH� GLIXVmR� VH� OLJDP� jV� SURWHtQDV� WHFLGXDLV� WRUQDQGR-se
LPXQRJrQLFRV�� (VVDV� SURWHtQDV� SRGHP� VHU� GHULYDGDV� GRV� TXHUDWLQyFLWRV��
FRPSRQHQWHV� GDV� FpOXODV� GH� /DQJHUKDQV� �&/�� RX� SHSWtGHRV� SUHYLDPHQWH�
processados ligados ao MHC.47�$�QDWXUH]D�GR�KDSWHQR��R�WLSR�GH�OLJDomR�GR�
hapteno com sHX� FDUUHDGRU� H� D� FRQILJXUDomR� WULGLPHQVLRQDO� ILQDO� GR�
complexo formado influenciam a imunogenicidade do complexo hapteno-
proteico.48
O complexo hapteno-SURWHLFR� p� FDSWDGR� SHODV� FpOXODV� GHQGUtWLFDV�
�&'��� IUDJPHQWDGR� HP� GLYHUVRV� SHSWtGLRV� H� DFRSODGRV� D� PROpFXODV� GR�
FRPSOH[R�SULQFLSDO�GH�KLVWRFRPSDWLELOLGDGH��0+&���2�FRQMXQWR�SHSWtGLR-MHC
p�H[SUHVVR�QD�VXSHUItFLH�GD�&'�SRVVLELOLWDQGR�D�DSUHVHQWDomR�GR�DQWtJHQR�
DR� OLQIyFLWR�7�KDSWHQR-HVSHFtILFR�QR� OLQIRQRGR� UHJLRQDO�48-50�$�FDSWDomR�GRV�
DQWtJHQRV�SURPRYH�XPD�VpULH�GH� DOWHUDo}HV�PRUIROyJLFDV�H� IXQFLRQDLV� QDV�
CD. Elas tornam-VH�PDLV�GHQGUtWLFDV��DXPHQWDm� R�Q~PHUR�GH�JUkQXORV�GH�
%LUEHFN�� GH� PROpFXODV� FR-HVWLPXODWyULDV� H� SURGX]HP� PDLRU� TXDQWLGDGH� GH�
citocinas. Dentre essas citocinas destacam-VH�DV�FLWRFLQDV�SUy-iQIODPDWyULDV�
IL-1 e TNF- .51,52
________________________________________________5HYLVmR�GD�OLWHUDWXUD 13
A IL-1 e o TNF- WUDQVIRUPDP� DV� &'� TXH� SDVVDP� GH� FpOXODV�
SUHSDUDGDV� SDUD� D� FDSWDomR� H� SURFHVVDPHQWR� GH� DQWtJHQRV� SDUD� FpOXODV�
HVSHFLDOL]DGDV� QD� DSUHVHQWDomR� DQWLJrQLFD�53 A IL-1 beta aumenta a
H[SUHVVmR�GH�PROpFXODV�FR-eVWLPXODWyULDV�FRPR�D�,&$0-1 e o CD86 nas CD,
QHFHVViULDV�SDUD�D�DWLYDomR�GRV�/7�KDSWHQR-HVSHFtILFRV�54,55 O TNF- atua
HP� GLYHUVRV� SRQWRV� QD� PLJUDomR� GDV� CD�� 'LPLQXL� D� H[SUHVVmR� GH� (-
caderinas nas CL�� XPD� PROpFXOD� GH� DGHVmR� DRV� TXHUDWLQyFLWRV�� LQGX]� D�
libeUDomR�GH�PHWDORSURWHLQDVHV��D�LQWHUDomR�GH�PROpFXODV�GH�DGHVmR��FRPR�
a LFA-1, ICAM-1 e VLA-6 das CD�FRP�D�PDWUL]�GpUPLFD��H�D�H[SUHVVmR�GH�
&&5��� XP� UHFHSWRU� TXH� UHVSRQGH� j� TXLPLRFLQDs� GR� WHFLGR� OLQIyLGH�
VHFXQGiULR�47,56-59 EVVDV�DOWHUDo}HV�levam�D�PLJUDomR das CD�HP�GLUHomR�DR�
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SURGX]LGDV� SRU�HVVDV�FpOXODV. $V�FLWRFLQDV�SUy-LQIODPDWyULDV�� ,/-1 e TNF- ,
DXPHQWDP� D� H[SUHVVmR� GH� (-selectinas e VCAM-�� QDV� FpOXODV�
endoteliais.60,61 A intHUDomR� HQWUH� R� VLDO\O� /HZLV� ;�� XPD� VHOHFWLQD� FXMa
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ligante, a E-VHOHFWLQD�GDV�FpOXODV�HQGRWHOLDLV�SURPRYH�D passagem das CD
SDUD�RV�OLQIiWLFRV�42,56,62,63
Dentro de vinte e quatro horaV�DSyV�R�FRQWDWR�FRP�R�DQWtJHQR��DV�
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GLYHUVRV� OLQIyFLWRV� 7� YLUJHQV� TXH� HVWmR� HP� SURFHVVR� GH� UHFLUFXODomR��3DUD
aWLYDU� DV� FpOXODV� 7� YLUJHQV� DV� &' devem passar dois sinais.65 Caso o
OLQIyFLWR� SRVVXD� R� UHFHSWRU� FRPSOHPHQWDU� DR� FRPSOH[R� SHSWtGHR-MHC ele
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UHFHEHUi� R� SULPHLUR� VLQDO�� 2� SULPHLUR� VLQDO� GHWHUPLQD� DOWHUDo}HV�
conformacionais nDV�PROpFXODV�FR-HVWLPXODWyULDV�GR�LT virgem tornando-as
mais iYLGDV� SHORV� VHXV� OLJDQWHV� TXH� HVWmR� SUHVHQWHV� QD� &'. Ainda, o
SULPHLUR�VLQDO�OHYD�D�WUDQVFULomR�GR�51$P�GD�,/-��SRUpP�R�51$P�IRUPDGR�p�
LQVWiYHO�66 2� VHJXQGR� VLQDO� p� GDGR� SHOD� OLJDomR� HQWUH� DV� PROpFXODV� FR-
HVWLPXODWyULDV� GDV� &D, ICAM-1, CD80 e CD86, com os seus respectivos
ligantes, LFA-1 e CD28 (que se liga tanto ao CD80 quanto ao CD86), no LT.
O CD86, quando acoplado ao CD28, estabiliza o RNAm da IL-2 induzindo o
/7� D� SURGX]LU� JUDQGH� TXDQWLGDGH� GHVVD� FLWRFLQD�� 2V� OLQIyFLWRV� DWLYados
passam a expressar o receptor completo da IL-2 tornando-VH�VXVFHWtYHLV�D�
esta citocina que� SRU� DomR� DXWyFULQD� OHYD� D� SUROLIHUDomR� FHOXODU�� SURFHVVR�
HVVH� FRQKHFLGR� FRPR� H[SDQVmR� FORQDO�66-68� $� H[SDQVmR� FORQDO� IRUPD� um
JUDQGH� Q~PHUR� GH� FpOXODV� 7� KDSWHQR-HVSHFtILFDV� TXH� UHVSRQGHUmR� HP� XP�
IXWXUR�FRQWDWR�FRP�R�DOpUJHQR. 1D�DWLYDomR�GDV�FpOXODV�7��DOpP�GR�SULPHLUR�H�
segundo sinais, as citocinas produzidas pelas CD e presentes no
PLFURDPELHQWH� RQGH� RFRUUH� D� DSUHVHQWDomR� DQWLJrQLFD� WDPEpP� WrP� XP�
papel essencial. AV� FLWRFLQDV� GHWHUPLQDP� R� WLSR� GH� UHVSRVWD� DR� DQWtJHQR�
apresentado, a IL-���OHYD�D�IRUPDomR�GH�FpOXODV�HIHWRUDV�DR�SDVVR�TXH�D�,/-
���GHWHUPLQD�R�VXUJLPHQWR�GH�FpOXODV�UHJXODGRUDV��FpOXODV�TXH�VXSULPHP�R�
processo.69,70 $SyV� D� H[SDQVmR� FORQDO nos linfonodos regionais, os LT
passam para R� GXFWR� WRUiFLFR e� HQWUDP� QD� FLUFXODomR� VDQJ�tQHD�� Essas
FpOXODV� 7� KDSWHQR-HVSHFtILFDV� H[SUHVVDP� R� DQWtJHQR� OLQIRFLWiULR� FXWkQHR�
�&/$�� TXH� GLUHFLRQD� HVVHV� OLQIyFLWRV� SUHIHUHQFLDOPHQWH� SDUD� RV� SURFHVVRV�
LQIODPDWyULRV�FXWkQHRV.
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3.2.2 Fase eferente
3DUD� HQFRQWUDU� R� DOpUJHQR�� DV� FpOXODV� 7� SUHFLVDP� SDVVDU� SHOD�
PLFURYDVFXODWXUD�GpUPLFD��SHOD�PDWUL]�GpUPLFD�H�DOFDQoDU�Rs TXHUDWLQyFLWRV
modificadoV�SHOR�DQWtJHQR� RQGH� DWXDUmR��7RGD�HVVD� SDVVDJHP�p� UHJXODGD�
SRU� TXLPLRFLQDV� H� PROpFXODV� GH� DGHVmR� H[SUHVVDV� QRV� WHFLGRV� H�
reconhecidas pelos LT.
2�/7� DWLYDGR� DSUHVHQWD� R� DQWtJHQR� GH� ³KRPLQJ´� �&/$��� R�9/$-4 e
receptores para quimiocinas.70-74�2�&/$�VH� OLJD� j�(-selectina expressa no
HQGRWpOLR� HVWLPXODGR� SHOD� SUHVHQoD� GR� DQWtJHQR� na pele sobrejacente
LQLFLDQGR� R� SURFHVVR� GH� URODJHP�� 3RUpP�� VRPHQWH� TXDQGR� R� 9/$-4 ou o
LFA-�� GRV� OHXFyFLWRV� VH� OLJDP� UHVSHFWLYDPHQWH� jV� LQWHJULQDV� HQGRWHOLDLV�
VCAM-1 e ICAM-1, forma-VH�XPD�OLJDomR�ILUPH�TXH�FRPELQDGD�DR�HVWtPXOR�
de quimiocinas permite a diapedese.60,75-77 &RPR�D�H[SUHVVmR�GH�,&$0-1 no
HQGRWpOLR�Vy�DXPHQWD����KRUDV�DSyV�R�FRQWDWR�FRP�R�DQWtJHQR��SHUtRGR�QR�
TXDO�JUDQGH�SDUWH�GR�LQIOX[R�GH�OLQIyFLWRV�Mi�RFRUUHX��D�(-selectina e o VCAM-
��SDUHFHP�VHU�SDUWLFXODUPHQWH�LPSRUWDQWHV�QR�LQtFLR�GR�SURFHVVR�e o ICAM-1
QD�VXD�DPSOLILFDomR�77
Uma vez na derme, o VLA-4 e o VLA-5 dos LT se ligam a
ILEURQHFWLQD�� XPD� SURWHtQD� H[WUDFHOXODU� GD� PDWUL]� GpUPLFD�� o que facilita o
WUkQVLWR�GHVVDV�FpOXODV�QHVVH�PHLR�78 $V�TXLPLRFLQDV�GLUHFLRQDP�RV�OLQIyFLWRV�
para o epitpOLR�H�D�OLJDomR�HQWUH�R�,&$0-���H[SUHVVR�QRV�TXHUDWLQyFLWRV��FRP�
o LFA-��GRV� OHXFyFLWRV�SURPRYH�D� LQWHUDomR�GHVVDV�FpOXODV�79�2V� OLQIyFLWRV�
SURGX]HP�XPD�UHVSRVWD�LQIODPDWyULD�YLJRURVD�SDUD�HOLPLQDU�RV�TXHUDWLQyFLWRV�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 16
PRGLILFDGRV� SHOR� DQWtJHQR�� /LEHUDP� JUDQde quantidade de IFN- que
HVWLPXOD� RXWUDV� FpOXODV� 7�� FpOXODV� 1.� H�PDFUyIDJRV� D� DPSOLDU� R� SURFHVVR�
LQIODPDWyULR�H�DXPHQWD�D�H[SUHVVmR�GH�)DV�QRs�TXHUDWLQyFLWRV� WRUQDQGR-os
PDLV� VXVFHWtYHLV� j� FLWRWR[LFLGDGH�PHGLDGD� SRU� )DV/�80,81� $OpP� GD� YLD� )DV-
FasL, foi dHPRQVWUDGR� TXH� DV�SHUIRULQDV� WDPEpP�SDUWLFLSDP�GD� GHVWUXLomR�
celular na dermatite de contato.82� 2V� TXHUDWLQyFLWRV� HQWUDP� HP� DSRSWRVH�
RFRUUHQGR�D�FOLYDJHP�GD�FDGHULQD�(�TXH�UHVXOWD�HP�SHUGD�GD�FRHVmR�FHOXODU�
GHPRQVWUDGD� SHOD� HVSRQJLRVH� H� IRUPDomR� GH� YHVtFXOas.83� $� GHVWUXLomR�
WHFLGXDO� H� D� GHVFDPDomR� UHPRYHm� R� DQWtJHQR� GR� WHFLGR�� GLPLQXLQGR� R�
processo�LQIODPDWyULR.42
3.2.3 &pOXODV�HQYROYLGDV
3.2.3.1 &pOXODV�GHQGUtWLFDV
���$� DSOLFDomR� GH� KDSWHQRV� QD� SHOH� LQGX]� D� H[WHQomR� H� UHWUDomR�
sucessiva dos dendritos dDV�FpOXODV�GH�/DQJHUKDQV��&/���EHP�FRPR�LQGX]�D�
PLJUDomR�GHVVDV�FpOXODV�SDUD�RV� OLQIRQRGRV�UHJLRQDLV�84 Esses movimentos
VmR� HVWLPXODGRV� SHOD� ,/-1 e TNF alfa, citocinas produzidas pelos
TXHUDWLQyFLWRV�H�SHODV�SUySULDV�&/�DSyV�R�FRQWDWR�FRP�R�DQWtJHQR�85 Essas
FLWRFLQDV� WDPEpP� LQGX]HP� D� PDWXUDomR� H� PLJUDomR� GDV� &/�86 Durante o
SURFHVVR�GH�DPDGXUHFLPHQWR�DV�&'�DXPHQWDP�D�H[SUHVVmR�GH�PROpFXODV�
FRHVWLPXODWyULDV�FRPR�R�&'����&'���H�&'����PROpFXODV�GH�DGHVmR�FRPR�D�
ICAM-1 e citocinas, como a IL-12.87-89 O processo de DPDGXUHFLPHQWR� p�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 17
QHFHVViULR� SDUD� D� DWLYDomR� GDV� FpOXODV� 7� KDSWHQR-HVSHFtILFDV� YLUJHQV� QRV�
/)1�UHJLRQDLV�SDUD�/7�HIHWRUHV�H�GH�PHPyULD�90 O amadurecimento descrito
RFRUUH�FRP�D�H[SRVLomR�DRV�KDSWHQRV��RV�LUULWDQWHV�TXDQGR�DSOLFDGRV�j�SHOH�
induzem a miJUDomR�GDV�&/��PDV�QmR�D�PDWXUDomR�� LPSHGLQGR�D�IRUPDomR�
GH� XPD� UHVSRVWD� HVSHFtILFD�89� $V� &'� HVWLPXODGDV� VmR� DWUDtGDV� SDUD� RV�
OLQIiWLFRV� DIHUHQWHV�� SRLV� SDVVDP� D� H[SUHVVDU� R� &&5�� TXH� UHVSRQGH� DV�
TXLPLRFLQDV� GR� WHFLGR� OLQIyLGH� &&/��� H� &&/���91� 2V� OLQIiWLFRV aferentes
expressam CCL21 e a zona paracortical dos linfonodos tanto CCL21 quanto
&&/��� DWUDLQGR� DV� &'� SDUD� HVWD� UHJLmR� Gos linfonodos pelo gradiente de
citocinas.92
Recentemente o papel das CD na DCA vem sendo revisto.
3.2.3.1.1 As CpOXODV�GHQGUtWLcas na fase aferente
+i� FRQWURYpUVLDV� TXDQWR� j� IXQomR� GDV� &/� QD� IDVH� GH� LQGXomR da
'&$��$�ODQJHULQD�p�XPD�SURWHtQD�WUDQVPHPEUDQD�TXH�OHYD�D�IRUPDomR�GRV�
JUkQXORV�GH�%LUEHFN��XP�PDUFDGRU�HVSHFtILFR das CL. Utilizando um modelo
PXULQR�RQGH�D�LQMHomR�GH�WR[LQD�GLIWpULFD�OHYD�D�GHSOHomR�VHOHWLYD�GDV�FpOXODV�
que expressam a langerina, Bennett et al.93 demonstraram em maio de 2005
TXH�D�DXVrQFLD�GDV�&/�GLPLQXL�D�FKDQFH�GH�VHQVLELOL]DomR�DRV�KDSWHQRV. Os
autores creditaraP� D� VHQVLELOL]DomR� GH� DOJXQV� GRV� FDPXQGongos expostos
no ensaio DV�&'�GpUPLFDV�H�FRQFOXtUDP�TXH�HVVDV�FpOXODV� trabalham junto
com as CL QR� SURFHVVR� GH� VHQVLELOL]DomR� H� TXH� D� DXVrQFLD� GHVWDV� DIHWD�
esse mecanismo.� 1R� HQWDQWR�� QR� PHVPR� PrV�� .LVVHQSIHQQLJ� et al.94
________________________________________________5HYLVmR�GD�OLWHUDWXUD 18
utilizando um modelo semelhante encontraram uma resposta aos haptenos
VHPHOKDQWH� HQWUH� FDPXQGRQJRV� FRP� GHSOHomR� GDV� &/� H� FRQWUROHV� H�
FRQFOXtUDP�TXH�DV�&/�VmR�GLVSHQViYHLV�SDUD�D�DSUHVHQWDomR�GRV�KDSWHQRV,
ILFDQGR�HVVD�IXQomR�D�FDUJR�DSHQDV�GDV�&'�GpUPLFDV. Em dezembro desse
mesmo ano, Kaplan et al.95 demonstraram que camundongos que
FRQVWLWXFLRQDOPHQWH� QmR� DSUHVHQWDP� &/� WrP� UHVSRVWD� DXPHQWDGD� DRV�
haptenos, ou seja, segundo esse modelo as CL possuem um papel
regulador. $Wp� HQWmR� R� SDSHO� GD� &/� QD� VHQVLELOL]DomR� IRL� GHILQLGR� FRPR�
indutor, indiferente ou supressor. Em 2007, Bennett et al.96 colaboradores
voltaraP� DR� VHX� PRGHOR� H� GHPRQVWUDP� TXH� RV� DQWtJHQRV� QmR� VmR�
DGHTXDGDPHQWH� WUDQVSRUWDGRV� SDUD� RV� OLQIRQRGRV� QD� DXVrQFLD� GDV� &/� H�
concluem que isso diminui a VHQVLELOL]DomR�DRV�DQWtJHQRV��FRQILUPando seus
achados no primeiro ensaio. AtributUDP� jV� GLIHUHQoDV� HQFRQWUDGDV� nos
outros estudos a: 1)� DXVrQFLD� SHUHQH� GDV� &/� QR� PRGHOR� GH� .DSODQ� H�
colaboradores H� ��� DR� XVR� GH� FRQFHQWUDo}HV� HOHYDGDV� GRV� DOpUJHQRV� QRV�
demais estudos. As CL em seu estado imaturo carregam auto-DQWtJHQRV�
SDUD� RV� OLQIRQRGRV� JHUDQGR� PHFDQLVPRV� GH� WROHUkQFLD� FRPR� FpOXODV� 7�
reguladoras.97.98� $� DXVrQFLD� GHVVDV� FpOXODV� SRGH� WHU� LPSHGLGR� R�
desenvolvimento desse mecanismo e levado a um estado de
KLSHUUHDWLYLGDGH��UHVSRQViYHO�SHOR�DXPHQWR�GD�VHQVLELOL]DomR�DRV�KDSWHQRV.
&RP� UHODomR� DR� HPSUHJR� GH� FRQFHQWUDo}HV� HOHYDGDV� GRV� DOpUJHQRV os
DXWRUHV� UHIHUHP� TXH� QmR� UHIOHWHP� DV� H[SRVLo}HV� KDELWXDLV�� PDV� VLP� D�
artificialidade dos modelos criados, o que pode ter gerado uma via de
resposta alternativa (Bennett 2007, 34 do Bennett 2007 e os 3 patetas).96,98
________________________________________________5HYLVmR�GD�OLWHUDWXUD 19
Mais tarde Fukunaga et al.99 demonstraram que camundongos com defeito
PLJUDomR� GDV� &/� SDUD� RV� OLQIRQRGRV� UHJLRQDLV��PDV� FRP�PLJUDomR� QRUPDO�
GDV�&'�GpUPLFDV��WHP�UHVSRVWD�QRUPDO�DRV�KDSWHQRV�H�VXJHUHm que as CD
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contra os haptenos do que as CL. Coletivamente, esses dados demonstram
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HIHWRUD�DRV�KDSWHQRV��PDV�QmR�SHUPLWHP�XPD�FRQFOXVmR�DGHTXDGD�TXDQWR�
ao papel das CL.
Recentemente um novo subtipo de CD langerina+ foi descrita, a CD
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OLQIRQRGRV� UHJLRQDLV��SRLV�VmR� RULXQGDV� GH� XPD� OLQKDJHP� FHOXODU�GLIHUHQte,
TXDQGR�GHSOHWDGDV�VmR�FDSD]HV�GH� UHSRYRDU�RV� WHFLGRV�PXLWR�PDLV� UiSLGR�
que as CL e apesar de serem fenotipicamente muito semelhantes
DSUHVHQWDP� PDUFDGRUHV� GH� VXSHUItFLH� SUySULRV.100-102� $V� &'� GpUPLFDV�
langerina+�WDPEpP�FDSWDP�H�DSUHVHQWDP�DQWtJHQRV��8Vando um modelo de
DEODomR� VHOHWLYD� GDV� &/� H� &'� GpUPLFDV� ODQJHULQD+, Wang et al.103
GHPRQVWUDUDP� TXH� D� WHQWDWLYD� GH� VHQVLELOL]DomR� LPHGLDWDPHQWH� DSyV� D�
GHSOHomR��PRPHQWR�TXH�VH�FDUDFWHUL]D�SHOD�DXVrQFLD� WDQWR�GDV�&/�TXDQWR�
GDV� &'� GpUPLFDV� OHQJHULQD+�� p� IUXVtra mas quando realizada alguns dias
DSyV� D� DEODomR�� TXDQGR� SDUWH� GDV� &'� GpUPLFDV� ODQJHULQD+� Mi� UHWRUQDUDP�
PDV�DV�&/�QmR��D�UHVSRVWD�D�VHQVLELOL]DomR�p�QRUPDO��(VVH�HQVDLR�DSRQWD�
SDUD�DV�&'�GpUPLFDV�ODQJHULQD+�H�QmR�SDUD�DV�&/�H�&'�GpUPLFDV�ODQJHULQD-
como�SULQFLSDLV�UHVSRQViYHLV�SDUD�R�GHVHQYROYLPHQWR�GD�'&$��1R�HQWDQWR��
Bursch et al.100 e Bennett et al.96 utilizaram um sistema semelhante e
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processo.
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em estudo.
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Gocinski et al.109� GHPRQVWUDUDP� TXH� FDPXQGRQJRV� FRP� GHSOHomR�
de LT CD8+ induzida pelo uso de anticorpo monoclonal anti-&'�� VmR�
incapazes de desenvolver DCA. No entanto, camundRQJRV� FRP� GHSOHomR�
induzida de LT CD4+� GHVHQYROYHP� UHVSRVWD� FOtQLFD� PDLV� LQWHQVD� H�
SURORQJDGD� DR� DOpUJHQR�� 5HVXOWDGRV� VHPHOKDQWHV� IRUDP� REWLGRV� FRP�
FDPXQGRQJRV�FRP�³NQRFNRXW´ �LQDWLYDomR��GRV�0+&�FODVVH�,�H�,,��$�DXVrQFLD�
GHVVDV� PROpFXODV� LPSHGH� D� DWLYDomR dos LT CD8+ e LT CD4+
UHVSHFWLYDPHQWH�� OHYDQGR� DV�PHVPDV� FRQVHT�rQFLDV� GD� DXVrQFLD� VHOHWLYD�
GHVVDV�VXESRSXODo}HV�GH�OLQIyFLWRV�110
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(P� XPD� DYDOLDomR� VHT�HQFLDO� GR� LQILOWUDGR� LQIODPDWyULR� Ga DCA
Okazaki et al.111� GHPRQVWUDUDP� TXH� RV� OLQIyFLWRV� HQFRQWUDGRV� QR� LQtFLR� GR�
SURFHVVR�VmR�/7�&'�+ produtores de IFN- seguidos de LT CD4+. A maior
SURSRUomR�GH�/7�&'�+ IRL�HQFRQWUDGD�DSyV����KRUDV�GR�FRQWDWR�H�GH�&'�+
DSyV����KRUDV�
Em 1998, Cavani et al.112� GHPRQVWUDUDP� TXH� DSHQDV� LQGLYtGXRV�
DOpUJLFRV�DR�QtTXHO�Dpresentam LT CD8+ (Tc1) DQWtJHQR-HVSHFtILFRV��3RUpP��
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DOpUJLFRV� H� QmR� DOpUJLFRV� GLIHULQGR� DSHQDV� QD�PDLRU� SURSRUomR� GH� FpOXODV�
supressoras, produtoras de IL-���� QR� JUXSR� VmR�70 A IL-10 inibe a
GLIHUHQFLDomR� H� PDWXUDomR� GDV� &'� EORTXHDQGR� D� OLEHUDomR� GH� ,/-12
QHFHVViULD�SDUD�JHUDU�XPD�UHVSRVWD�DOpUJLFD�112
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o LT CD8+�p�SRVVtYHO�TXH�D�QDWXUH]D�GR�DQWtJHQR�H�RX�D�YLD�GH�DFHVVR�GR
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UHVSRVWD�TXH�VH�IRUPDUi�113
���$OpP� GRV� /7� &'�+ e LT CD4+ produtores de IFN- , os LTh17
WDPEpP�H[HUFHP�SDSHO�HIHWRU�LPSRUWDQWH�QD�'&$��4XDQGR�HVWLPXODGRV�SHOR�
contato com os haptenos RV� TXHUDWLQyFLWRV�KXPDQRV� SURGX]HP� ,/-23 que,
junto com a IL-1 beta, leva ao desenvolvimento de LTh17.114�2V�LQGLYtGXRV�
FRP� VHQVLELOLGDGH� GH�FRQWDWR�DSUHVHQWDP�/7K��� QR� VDQJXH� SHULIpULFR� TXH�
UHVSRQGHP�DV�FpOXODV�DSUHVHQWDGRUDV�GH�DQWtJHQR�TXH�SRUWDP�R�DOpUgeno.114
(VVDV� FpOXODV� VmR� CD4+ ou CD8+ e produzem IL-17. A IL-17 afeta os
TXHUDWLQyFLWRV� GH� GLYHUVDV� IRUPDV�� HVWLPXOD� D� SURGXomR� GH� FLWRFLQDV� SUy-
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LQIODPDWyULDV�� PRGXOD� D� OLEHUDomR� GH� TXLPLRFLQDV�� SHOR� DXPHQWR� GD�
H[SUHVVmR� GH� ,/-�� H� GLPLQXLomR� GH� 5$17(6� H� DXPHQWD� D� H[SUHVVmR� GH�
ICAM-���TXH�IDYRUHFH�D�LQWHUDomR�FRP�RV�OLQIyFLWRV�115 Dessa forma a IL-17
DPSOLD� R� SURFHVVR� LQIODPDWyULR� ORFDO�115-117 Um modelo experimental
GHPRQVWURX� TXH� D� DXVrQFLD� GD� ,/-17 em camundongos compromete o
GHVHQYROYLPHQWR�GD�UHDomR�Ge hipersensibilidade de contato.118
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efetora da DCA por dinitrofluorobenzeno em camundongos com ³NQRFN-RXW´
GR� JHQH�5DJ���� JHQH� HVVHQFLDO� SDUD� D�GHVHQYROYLPHQWR� GRV� OLQIyFLWRV� 7� H�
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XPD�UHVSRVWD�QmR�FRQWURODGD do sistema imune aos haptenos.120 O controle
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atividade reguladora.121� $� SHUGD� GHVVHV� PHFDQLVPRV� GH� WROHUkQFLD� OHYD� D�
DCA.
O conhecimento sobre DV�FpOXODV�7�UHJXODGRUDV�vem sendo revisto
QRV� ~OWLPRV� DQRV�� (VVDV� FpOXODV� VmR� XPD� VXEIDPtOLD� KHWHURJrQHD� GH�
OLQIyFLWRV� 7� TXH� VXSULPH� D� UHVSRVWD� LPXQH� SHOD� OLEHUDomR� GH� FLWRFLQDV�
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UHJXODGRUDV����7U���H�DV�FpOXODV�7UHJ�&'�+CD25+��$V�FpOXODV�7U��SURGX]HP�
grande quantidade de IL-10, moderada quantidade de IL-5 e TGF- � H� QmR�
produzem IL-4 e IFN- ��³,Q�YLWUR´�HVVDV�FpOXODV�UHVWULQJHP�D�GLIHUHQFLDomR�H�
SURGXomR�GH�,/-���SHODV�&'�LQLELQGR�D�DSUHVHQWDomR�DQWLJrQLFD�70,123 Esses
HIHLWRV�VmR�PHGLDGRV�SHOD�,/-���H�UHVXOWDP�QD�VXSUHVVmR�GDV�FpOXODV�7�&'�+
e CD8+ hapteno-HVSHFtILFDV�123 Cavani et al.112 demonstraram que os LT
CD4+� SHULIpULFRV� GRV� LQGLYtGXRV� QmR� DOpUJLFRV� DR� QtTXHO� H[SUHVVDP� PDLRU�
quantidade de IL-10 e menor de IFN- �TXH�RV�SDFLHQWHV�DOpUJLFRV��RX�VHMD��
LQGLYtGXRV� QmR� DOpUJLFRV� DSUHVHQWDP� PDLRU� TXDQWLGDGH� GH� FpOXODV� 7U��
hapteno-HVSHFtILFDV�QR�VDQJXH��$V�FpOXODs Tr1 hapteno-HVSHFtILFDV�WDPEpP�
SRGHP�VHU�HQFRQWUDGDV�QDV�OHV}HV�GH�SHOH�GD�VHQVLELOLGDGH�GH�FRQWDWR�RQGH�
modulam a intensidade do processo.70,123
���2� VHJXQGR� WLSR� GH� FpOXOD� UHJXODGRUD� EHP� HVWXGDGD� H[SUHVVD� D�
PROpFXOD�&'��� D�FDGHLD� DOID� GR� UHFHSWRU� GD� ,/-�� �&'����� R� DQWtJHQR��� GR�
OLQIyFLWR�7�FLWRWy[LFR��&7/$-���H�R�IDWRU�GH�WUDQVFULomR�)R[S���(VVDV�FpOXODV�
VmR� FKDPDGDV� GH� /7UHJ� &'�+CD25+ ou Tregs.124� (ODV� WDPEpP� SRGHP�
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H[SUHVVDU�R�&/$��SUHVXPLYHOPHQWH�DSyV�R�HQFRQWUR�FRP�&'�QRV�OLQIRQRGRV�
regionais, e migrar para a pele.125� (VVDV� FpOXODV� HVWmR� HQYROYLGDV� QD�
WROHUkQFLD�DR�SUySULR�H�WDPEpP�VmR�FDSD]HV�GH�LQLELU�D�DWLYDomR�³LQ�YLWUR´�GDV�
FpOXODV�HIHWRUDV�H[SRVWDV�DR�VHQVLELOL]DQWH�GH�LQGLYtGXRV�QmR�DOpUJLFRV��PDV�
QmR�GH� SDFLHQWHV� DOpUJLFRV�126,127 O mecanismo GD� VXSUHVVmR� LQGX]LGD�SRU�
HVVDV� FpOXODV� p� PRWLYR� GH� GLVFXVVmR�� (VWXGRV� ³LQ� YLWUR´� VXJHUHP� D�
QHFHVVLGDGH� GR� FRQWDWR� FpOXOD-FpOXOD� SHOD� LQWHUDomR� HQWUH� R� &7/$-4 da
FpOXOD� UHJXODGRUD�FRP�R�&'���H�&'���SDUD� LQDWLYDomR�GR�/7� HIHWRU�128� -i�
PRGHORV� ³LQ� YLYR´� DSRQWDP� SDUD� XPD� VXSUHVVmR� PHGLDGD� SHOD� DomR� GH�
citocinas, em particular a IL-10.129
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GH�FRRSHUDomR��IRL�GHPRQVWUDGR�TXH�DV�FpOXODV�7UHJ�LQGX]HP�D�SURGXomR�GH�
IL-���HP�FpOXODV�7U��130 Esses dois�WLSRV�GH�FpOXODV�UHJXODGRUDV�DSUHVHQWDP�
uma vasta gama de receptores de quimiocinas, como o CCR4 e CCR8, e
VmR� DWUDtGDV� SRU� TXLPLRFLQDV� SURGX]LGDV� QD� IDVH� WDUGLD� GD� '&$� FRPR� D�
CCL1.131,132 Dessa forma, atuam tanto na fase aferente quanto na fase
eferente, impedindo o surgimento da alergia e minimizando a intensidade e
GXUDomR�GR�SURFHVVR�TXDQGR�HOH�Mi�VH�GHVHQYROYHX�
$�PRELOL]DomR�H�PDWXUDomR�GDV�&'�p�SURPRYLGD�SHOD�H[SRVLomR�j�
VLQDLV� GH� SHULJR� FRPR� OHVmR� FHOXODU�� UDGLDomR� 89%�� EDFWpULDV� H� SURGXWRV�
virais.120� 2� HVWDGR� GH�PDWXUDomR� GDV� &' determina a capacidade dessas
FpOXODV� HP�direcionar os LT hapteno-HVSHFtILFRV�YLUJHQV� SDUD�LT hapteno-
HVSHFtILFRV�efetores, de PHPyULD�RX�VXSUHVVRUHV�121�&pOXODV�7�&'�+ virgens
quando estimulados com CD imaturas na presHQoD� GH� 7*)- se
________________________________________________5HYLVmR�GD�OLWHUDWXUD 26
transformam em LTreg CD4+CD25+.133�$�GLIHUHQFLDomR�SDUD�/7K��RX�LTc1
GHSHQGH� GD� SURGXomR� GH� ,/-12 pela CD, o que ocorre apenas quando a
PDWXUDomR�p�FRPSOHWD�121,134 A coH[LVWrQFLD�de sinais de perigo e H[SRVLomR�
aos� TXtPLFRs parece ser um IDWRU� LPSRUWDQWH� QD� SHUGD� GD� WROHUkQFLD� DRV�
haptenos. Dessa forma o efeito irritante��XPD�FDUDFWHUtVWLFD�GRV�DOpUJHQRV�GH�
contato mais sensibilizantes, pode ajudar a romper o mecanismo de
WROHUkQFLD� H� MXQWR� FRP� R� DOpUJHQR� GHWHUPLQDU� R� DPDGXUHFLPHQWR� FRPSOHto
das CD e induzir o desenvolvimento da DCA.135,136
$OpP� GHVVHV� PHFDQLVPRV� FRQWURODGRUHV�� *RUEDFKHY� et al. 137
GHPRQVWUDUDP�TXH�Ki�DSRSWRVH�GDV�FpOXODV�GHQGUtWLFDV�QRV�OLQIRQRGRV�H�TXH�
HVVH� PHFDQLVPR� WDPEpP� VXSULPH� D� '&$�� 2� DXWRU� GHPRQVWURX� TXH�
camundoQJRV� FRP� GHSOHomR� GH�FpOXODV� &'�+ ou com ³NQRFNRXW´ do gene
UHVSRQViYHO�SHOR�)DV/�DSUHVHQWDP�PDLRU�WHPSR�GH�SHUPDQrQFLD�GDV�FpOXODV�
GHQGUtWLFDV�QRV�OLQIRQRGRV�HP�UHODomR�DRV�FDPXQGRQJRV�YLUJHQV��$�SHUGD�GD�
DSRSWRVH� GDV� FpOXODV� DSUHVHQWDGRUHV� UHVXOWRX� HP� DWLYDomR� LQWHQVD� H�
sustentada de LT CD8+ produtores de IFN- nos camundongos modificados.
$OpP� GHVVHV� DFKDGRV� ODERUDWRULDLV�� RV� FDPXQGRQJRV�� FRP� GHSOHomR� GH�
FpOXODV�&'�+�RX�FRP�DOWHUDomR�GR�)DV/��DSUHVHQWDUDP�UHVSRVWD�FOtQLFD�PDLV�
intensa e persistente ao alpUJHQR�WHVWDGR�
3.2.3.3 4XHUDWLQyFLWRV
���6mR� FpOXODV� FUtWLFDV� QD� UHVSRVWD� LPXQH� GD� SHOH� GHYLGR� D� VXD�
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quanto no controle da resposta aos haptenos. Os receptores para IL-1 dos
queratiQyFLWRV� UHVSRQGHP�D� ,/-1 � OLEHUDGD�SHODV� &/� H[SRVWDV�DR� DQWtJHQR�
produzindo TNF- � TXH� UHVXOWD� QD� PDWXUDomR� H� PLJUDomR� GDV� &/�138,139 O
IFN- � SURGX]LGR� SHORV� /7� DXPHQWD� D� H[SUHVVmR� GH� ,&$0-1 nos
TXHUDWLQyFLWRV� TXH� VH� OLJD� DR� /)$-�� GRV� OLQIyFLWRV� IDFLOLWDQGR� D� LQILOWUDomR�
GHVVDV� FpOXODV� SDUD� D� HSLGHUPH� H� R� SURFHVVR� GH� DSUHVHQWDomR�
DQWLJrQLFD�140,141�$OpP�GLVVR��R�,)1- �DXPHQWD�D�H[SUHVVmR�GH�PROpFXODV�GR�
0+&�FODVVH� ,,� QRV�TXHUDWLQyFLWRV�141�'HVVD� IRUPD�RV�TXHUDWLQyFLWRV�SRGHP�
DSUHVHQWDU�R�DQWtJHQR�SDUD�RV�/7&D8+, pois constitucionalmente expressam
MHC classe I e para os LTCD4+�� SRLV� VmR� LQGX]LGRV� D� H[SUHVVDU� R� 0+&�
classe II pelo IFN- �� (P� FRQGLo}HV� QRUPDLV� RV� TXHUDWLQyFLWRV� H[SUHVVDP�
EDL[RV� QtYHLV� GDV� PROpFXODV� FRHVWLPXODWyULDV� &'��� H� &'���45 Essas
PROpFXODV� VH� OLJDP� DRV� VHXV� UHFHSWRUHV� QD� FpOXOD� 7� �&'���&7/$-4 ±
DQWtJHQR-�� DVVRFLDGR�DR� /7�FLWRWy[LFR�� H�VmR�QHFHVViULDV� SDUD� OHYDU� D�XP�
segundo sinal efetivo.45�1D�DXVrQFLD�GH�XP�VHJXQGR�VLQDO�RV�/7�WRUQDP-se
DQpUJLFRV�142,143� (VVHV� /7� DQpUJLFRV� H[SUHVVDP� JUDQGH� Tuantidade de
receptores para IL-�� H� SRUWDQWR�FRPSHWHP�FRP�DV� FpOXODV�7� HIHWRUDV� H� GH�
PHPyULD� SRU� HVVH� IDWRU� GH�FUHVFLPHQWR��2� FRQWDWR�GRV� TXHUDWLQyFLWRV�FRP�
DOpUJHQRV�H�LUULWDQWHV� ID]�FRP�TXH�RV�TXHUDWLQyFLWRV�KXPDQRV�DXPHQWHP�D�
H[SUHVVmR�GH�&'���IDYRUHFHQGR�R�GHVHQYROYLPHQWR�GD�UHVSRVWD�DOpUJLFD�GH�
contato.144�$OpP�GLVVR�� FRPR�YLVWR��RV� TXHUDWLQyFLWRV� SURPRYHP�D� JHUDomR�
GH� /7K��� SHOD� SURGXomR� GH� ,/-1 beta e IL-23 que ampliam o processo
LQIODPDWyULR�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 28
���3RU�RXWUR� ODGR�RV�TXHUDWLQyFLWRV�VXSULPHP�D�'&�SHOD�SURGXomR�GH�
IL-��� HP� UHVSRVWD� D� H[SRVLomR� DRV� KDSWHQRV�145,146� $� H[SRVLomR� DRV�
DOpUJHQRV� WDPEpP� LQGX]� D� SURGXomR� GH� ,/-��� TXH� HVWi� HQYROYLGD� QD�
quimiotaxia de LT CD4+� TXH� VXSULPHP� D� UHVSRVWD� LQIODPDWyULD�147� $OpP�
GLVVR�� RV� TXHUDWLQyFLWRV�� HP� XP� DPELHQWH� LQIODPDWyULR�� H[SUHVVDP� DOWRV�
QtYHLV�GR�OLJDQWH�GR�UHFHSWRU�DWLYDGRU�GR�IDWRU�QXFOHDU� B (RANKL) que induz
D�H[SUHVVmR�GH�&'����H�&'���QDV�&/�TXDQGR�VH�OLJD�DR�UHFHSWRU�DWLYDGRU�
do fator nuclear B (RANK).148� $� H[SUHVVmR� GH� &'���� HVWi� DVVRFLDGD� j�
LQGXomR�GH�Fplulas CD4+CD25+ que suprimem a resposta imune.149
3.2.3.4 0DVWyFLWRV
���-XQWR�FRP�RV�TXHUDWLQyFLWRV�H�FpOXODV�HQGRWHOLDLV�RV�PDVWyFLWRV�VmR�
fonte importante de TNF- e atuam tanto na fase aferente quanto eferente
da hipersensibilidade de contato.150 O TNF- �p�LPSRUWDQWH�QD�PDWXUDomR�GDV�
&'�H�SDVVDJHP�GDV�PHVPDV�SHOR�HQGRWpOLR� 7DPEpP�SURPRYH�D�LQILOWUDomR�
GH� FpOXODV� 7� DXPHQWDQGR� D� UHDomR� LQIODPDWyULD�� $VVLP� FRPR� RV�
TXHUDWLQyFLWRV��RV�PDVWyFLWRV� WrP�IXQomR�GXSOD��SRLV�VXSULPHP�D�'&$�SHOD�
SURGXomR�GH�,/-10.151
3.2.3.5 /LQIyFLWRV�%�H�FpOXODV�7�1.
A DCA foi considerada por muito tempo um processo independente
GD�SDUWLFLSDomR�GDV�FpOXODV�%��PDV�HVWXGRV�UHFHQWHV�DSRQWDP�SDUD�XP�SDSHO�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 29
HVVHQFLDO�GHVVDV�FpOXODV��2V�OLQIyFLWRV�%��/%��DVVRFLDGRV�j�'&$�VmR�GR�WLpo
�� TXH� VmR� FpOXODV� %� LQGHSHQGHQWHV� GDV� FpOXODV� 7�� QmR� IRUPDP� FHQWURV�
JHUPLQDWLYRV��JHUDOPHQWH�QmR�SDVVDP�SHOR�UHDUUDQMR�GR�'1$�H�VmR�IRQWH�GH�
IgM.152�&DPXQGRQJRV�FRP�GHSOHomR�GH�/%�WLSR���DSUHVHQWDP�GLPLQXLomR�GD�
'&$� TXH� p� UHVWDXUDGD� FRP� D� UHSRVLomR� GD� ,J0� PRQRFORQDO� DQWtJHQR-
HVSHFtILFD�� SHOD� WUDQVIHUrQFLD� GH� /%� WLSR� �� GH� GRDGRUHV� DOpUJLFRV� DR�
DQWtJHQR�RX�GH�VRUR�GH�FDPXQGRQJRV����KRUDV�DSyV�D�VHQVLELOL]DomR�GHVWHV�
FRP�R�DQWtJHQR�HP�TXHVWmR�153�(VVD�,J0�FOLYD�D�IUDomR�&��GR�FRPSOHPHQWR�
formando o C5a que DWLYD� RV� PDVWyFLWRV� FRP� FRQVHT�HQWH� OLEHUDomR� GH�
TNF- �H�TXLPLRWD[LD�GRV�OHXFyFLWRV�154�&DPXQGRQJRV�FRP�GHILFLrQFLD�GH�&��
DSUHVHQWDP� GLPLQXLomR� QD� UHVSRVWD� GH� FRQWDWR� DRV� KDSWHQRV�155-157 A IgM
DQWtJHQR-HVSHFtILFD�p�SURGX]LGD�GXUDQWH�D�IDVH�DIHUHQWH�GD�'&$ onde os LB
tipo 1 proliferam rapidamente.153
���2V� /%� WLSR� �� SRU� VXD� YH]� VmR� DWLYDGRV� SHODV� FpOXODV� 7� 1.�� XP�
VXEWLSR� GH� OLQIyFLWR� TXH� ID]� SDUWH� GR� VLVWHPD� LPXQH� LQDWR�� $SHVDU� GH�
DSUHVHQWDUHP�UHFHSWRU�GH�FpOXODV�7�HVVDV�FpOXODV�QmR�SDVVDP�SRU�UHDUUDQMR�
JrQLFR� H� VmR� FDSD]HV� GH� VH� OLJDU�� DWUDYpV� GHVVH� 7&5�� D� JOLFROtSLGHV�
DOWDPHQWH� FRQVHUYDGRV� DFRSODGRV� DV� PROpFXODV� &'�G�� XPD� PROpFXOD�
VHPHOKDQWH� j� PROpFXOD� GR� 0+&� FODVVH� ,�� HQFRQWUDGD� QDV� FpOXODV�
DSUHVHQWDGRUDV�GH�DQWtJHQR�45,158,159�$�QDWXUH]D�GHVVH�JOLFROtSLde permanece
GHVFRQKHFLGD��$SyV�R�FRQWDWR�FRP�HVVH�JOLFROtSLGH�D�FpOXOD�7�1.�VH�SUROLIHUD�
QR�ItJDGR�H�OLEHUD�FLWRFLQDV�FRPR�D�,/-��TXH�p�UHVSRQViYHO�SHOD�DWLYDomR�GRV�
/%�WLSR���QD�SUHVHQoD�GR�DQWtJHQR�46,160
________________________________________________5HYLVmR�GD�OLWHUDWXUD 30
4 7HVWHV�GLDJQyVWLFRV
4.1 Teste de contato
2� GLDJQyVWLFR� HWLROyJLFR� GDV� '&� DOpUJLFDV� p� FRQILUPDGR� SHOR� TC,
considerado H[DPH� SDGUmR� RXUR� SDUD� D� LGHQWLILFDomR� GR� DJHQWH� FDXVDO��
Todavia,� HVVH� WHVWH� WHP� XPD� VpULH� GH� GHVYDQWDJHQV�� 2� WHVWH� HVWi�FRQWUD-
LQGLFDGR�QRV�SDFLHQWHV�����FRP�GRHQoD�DWLYD�����TXH fizeram uso recente de
FRUWLFRVWHUyLGH�VLVWrPLFR�RX�WySLFR�QR�ORFDO�GR�WHVWH�����TXH�IRUDP�H[SRVWRV�j�
UDGLDomR�XOWUDYLROHWD�QR� ORFDO�GR� WHVWH�H����HP�JHVWDomR��+i�D�QHFHVVLGDGH�
GH� PDWHULDO� DGHTXDGR�� FRQKHFLPHQWR� H� H[SHULrQFLD� TXDQWR� j� WpFQLFD� GH�
montagem H�DSOLFDomR��FRPSUHHQVmR�H�FRRSHUDomR�SRU�SDUWH�GR�SDFLHQWH�H�
tempo, tanto do profissional quanto do paciente.21 $LQGD��D� LQWHUSUHWDomR�p�
VXEMHWLYD� H� R� WHVWH� QmR� p� LQyFXR� podendo levar a� FRPSOLFDo}HV� ORFDLV� H�
VLVWrPLFDV� GH� JUDYLGDGH� YDULiYHO�� (QWUH� DV� UHDo}HV� ORFDLV� GHVFULWDV�
encontram-VH�� GHUPDWLWH� GH� FRQWDWR� LUULWDWLYD�� H[DFHUEDomR� GD� GHUPDWLWH�GH�
FRQWDWR�� VtQGURPH� GD� SHOH� H[FLWDGD�� UHDo}HV� SHUVLVWHQWHV�� SLJPHQWDomR� H�
FLFDWUL]HV��'HQWUH�DV�DGYHUVLGDGHV�VLVWrPLFDV�SRGHP�RFRUUHU��VHQVLELOL]DomR�
ativa, absoromR� GH� VXEVWkQFLDV� Wy[LFDV� H�RX� FDUFLQRJrQLFDV� H� UHDo}HV�
DQDILODFWyLGHV�2,21,161,162
Outro problema associado ao TC se refere j� UHSURGXWLELOLGDGH dos
resultados. O maior estudo sobre reprodutibilidade do TC foi feito pelo grupo
DOHPmR� GH� SHVTXLVD� HP� Germatite de contato que encontrou 15,1% de
GLVFRUGkQFLD�HQWUH�RV�UHVXOWDGRV�163
________________________________________________5HYLVmR�GD�OLWHUDWXUD 31
$OpP�GLVVR��R�TC com metais pode levar a respostas falso-positivas,
de natureza irritativa.16�1RV�(VWDGRV�8QLGRV�D�FRQFHQWUDomR�GR�ELFURPDWR�GH�
SRWiVVLR�QR�TC foi reduzida GH������SDUD�������SDUD�GLPLQXLU�R�Q~PHUR�GH�
UHVSRVWDV� LUULWDWLYDV��1R�HQWDQWR��DOJXQV�DXWRUHV� UHIHUHP�TXH�D�UHGXomR�GD�
FRQFHQWUDomR� SRGH� OHYDU� D� GLPLQXLomR� GD� VHQVLELOLGDGH� H� SRU� LVVR�
UHFRPHQGDP�WHVWDU�FRP������GH�ELFURPDWR�GH�SRWiVVLR�38 Na Europa esse
agente continua sendo testado a 0,5% tomando-VH� FXLGDGR� TXDQWR� j�
UHOHYkQFLD�GRV�UHVXOWDGRV�11
$� LPSRUWkQFLD� GR� GLDJQyVWLFR� HWLROyJLFR� QR� SURJQyVWLFR� GD� '&$�
DOLDGR� DV� OLPLWDo}HV� H� GLILFXOGDGHV� JHUDGDV� SHOR� 7&� DSRQWDP� SDUD� D�
necessidade do desenvolvimento GH� RXWUR� WHVWH� GLDJQyVWLFR�� $� DOWHUQDWLYD�
PDLV�SURPLVVRUD�p�R�WHVWH�GH�SUROLIHUDomR�OLQIRFLWiULD�27
4.2 Teste de pUROLIHUDomR�lLQIRFLWiULD
2�WHVWH�GH�SUROLIHUDomR�OLQIRFLWiULD��73/��YHP�VHQGR�HVWXGDGR�FRP�
R� SURSyVLWR� GH� FRPSOHPHQWDU� H� HYHQWXDOPHQWH� Vubstituir o TC. O TPL
FRPSDUD� D� UHVSRVWD� GRV� OLQIyFLWRV� HP� FXOWXUD� QD� SUHVHQoD� H� DXVrQFLD� GH�
PLWyJHQRV�RX�DQWtJHQRV��2V�PLWyJHQRV�VmR�DWLYDGRUHV�SROLFORQDLV�DR�SDVVR�
TXH� RV� DQWtJHQRV� DWLYDP� DSHQDV� RV� OLQIyFLWRV� TXH� SRVVXHP� R� UHFHSWRU�
DQWtJHQR� HVSHFtILFR� DGHTXDGR�� 4XDQGR� RV� OLQIyFLWRV� VmR� DWLYDGRV� SDVVDP�
SRU�DOWHUDo}HV�PRUIROyJLFDV��IXQFLRQDLV�H�SUROLIHUDP��Aumentam de tamanho,
DSUHVHQWDP� FLWRSODVPD�DPSOR� H� QXFOpROR�HYLGHQWH� WRUQDQGR-se semelhante
jV�FpOXODV�EOiVWLFDV��(VVH�IHQ{PHQR�p�GHQRPLQDGR�³WUDQVIRUPDomR�EOiVWLFD´�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 32
H� p� DFRPSDQKDGR� GR� DXPHQWR� GD�H[SUHVVmR�GH� FLWRFLQDV� H� GD�VtQWHVH� GR�
iFLGR�GHVR[LUULERQXFOpLFR��'1$���$�DWLYDomR�OLQIRFLWiULD�SRGH�VHU�TXDQWLILFDGD�
SHOD�GHWHUPLQDomR�SRU�PLFURVFRSLD�GD�SRUFHQWDJHP�GH�FpOXODV�EOiVWLFDV�QD�
FXOWXUD��SHOD�VtQWHVH�GH�'1$�GHVVDV�FpOXODV��TXH�SRGH�VHU� LQIHULGD�DWUDYpV�
GD� DGLomR� GH� WLPLGLQD� UDGLRPDUFDGD� QD� FXOWXUD� H� SRVWHULRU� GHWHFomR� GR�
PDWHULDO� UDGLRDWLYR� LQFRUSRUDGR�RX� SHOD� GHWHFomR�GDV�FLWRFLQDV�SURGX]LGDV��
,QGHSHQGHQWH�GR�PpWRGR��DV�FpOXODV�VmR�FXOWLYDGDV�QD�SUHVHQoD�H�DXVrQFLD�
GR�DQWtJHQR�H�D�DIHULomR�p�IHLWD�FRPSDUDQGR�RV�UHVXOWDGRV�HQFRQWUDGRV�HQWUH�
as culturas� HVWLPXODGDV� H� QmR� HVWLPXODGDV�28,66,164 $� GLIHUHQoD� H� D� UD]mR�
HQFRQWUDGDV� HQWUH� DV� FXOWXUDV� HVWLPXODGDV� H� QmR� HVWLPXODGDV�� FRQKHFLGDV�
como delta ( ) e tQGLFH� GH� HVWLPXODomR� �,(� respectivamente, VmR� RV�
SDUkPHWURV�PDLV�XWLOL]DGRV�FRPR�FULWpULR�GH�UHVSRVWD�DR�HVWtPXOR�164
���2�73/� Mi�SURYRX�VHU� ~WLO� QR� GLDJQyVWLFR� GH�'&$�SRU� QtTXHO�e vem
sendo estudado� SDUD� GLDJQyVWLFR� GH� KLSHUVHQVLELOLGDGH� DR� FURPR� GHVGH�
meDGRV�GR�VpFXOR�;; sem o mesmo sucesso. 24,25
Em 1966, Grosfeld et al.165 testaram o TPL em 5 pacientes com
'&$�SRU�FURPR�H���FRQWUROHV��$V�FXOWXUDV� IRUDP� UHDOL]DGDV�FRP� OHXFyFLWRV�
WRWDLV�HVWLPXODGRV�FRP�ELFURPDWR�GH�SRWiVVLR�QD�FRQFHQWUDomR�GH�294, 29,4
e, em um caso, 2,94 g/mL durante 3 a 4 dias. A leitura do teste foi feita pelo
SHUFHQWXDO�GH�FpOXODV�EOiVWLFDV��2�SUySULR�DXWRU�GHILQLX�VHXV�UHVXOWDGRV�FRPR�
QmR�FRQYLQFHQWHV�H�TXHVWLRQD�VH�R�XVR�GH� ILWRKHPDJOXWLQLQD��XP�PLWyJHQR��
SDUD�D�VHSDUDomR�GDV�FpOXlas e/ou o tempo de cultura, possivelmente curto,
WHULDP�VLGR�UHVSRQViYHLV�SHORV�UHVXOWDGRV��
________________________________________________5HYLVmR�GD�OLWHUDWXUD 33
Em 1971, Gerd Lischka166 XVRX� ELFURPDWR� GH� SRWiVVLR� QD�
FRQFHQWUDomR� ILQDO� GH� 0,294 e 2,940 g/mL em culturas de 5 dias de
OHXFyFLWRV� WRWDLV� GH� �� SDFLHQWHV� FRP� 'CA por cromo e 5 controles. Sete
SDFLHQWHV�H���FRQWUROH�UHVSRQGHUDP�DR�WHVWH��2V���SDFLHQWHV�UHVSRQGHUDP�j�
FRQFHQWUDomR�GH�2,940 g/mL H���SDFLHQWHV�H�XP�FRQWUROH�j�FRQFHQWUDomR�GH�
0,294 g/mL. Assim como Grosfeld e colaboradores analisou a resposta pelo
perFHQWXDO�GH�FpOXODV�EOiVWLFDV��XP�PpWRGR�WUDEDOKRVR�H�VXEMHWLYR e por isso
muito pouco utilizado hoje.
Em 1979, Christiansen et al. 167 o resultado do TPL para cromo,
QtTXHO�H�FREDOWR�HQWUH����SDFLHQWHV�FRP�SHUGD�DVVpSWLFD�GH�SUyWHVH�PHWiOLFD�
e 11 controOHV�FRP�SUyWHVH�EHP�VXFHGLGD�SRU�SHOR�PHQRV�GRLV�DQRV��3DUD�
WDQWR� HVWLPXODUDP� RV� OLQIyFLWRV� FRP� FORUHWR� GH� FURPR� QDV� FRQFHQWUDo}HV�
finais de 0,2665; 2,665; 26,65 e 266,5 g/mL em culturas de 6 dias. Os
resultados obtidos com o TPL para o cromo foram significativamente
VXSHULRUHV� QR� JUXSR� FRP� SHUGD� GH� SUyWHVH�� 4XLQ]H� FDVRV� H� DSHQDV� um
FRQWUROH� IRUDP� HVWLPXODGRV� �,(!���� VXJHULQGR� D� SDUWLFLSDomR� GD�
KLSHUVHQVLELOLGDGH� GRV� FRPSRQHQWHV� GD� SUyWHVH� QD� SHUGD� GDV� PHVPDV��
Esses autores encontraram os melhores resultados do TPL para o cromo
com�D�FRQFHQWUDomR�GH������ g/mL.
Em 1985, Carando et al.168 utilizaram o TPL em dois pacientes com
SHUGD�DVVpSWLFD�GH�SUyWHVH�PHWiOLFD��2V�DQWtJHQRV�XWLOL]DGRV�IRUDP�R�FORUHWR�
GH� FURPR�� FORUHWR� GH� QtTXHO� H� FORUHWR� GH� FREDOWR�� 2� FUomo foi testado na
FRQFHQWUDomR�GH����� J�PO�HP�FXOWXUDV�GH�OLQIyFLWRV�GH�DSHQDV����KRUDV��2V�
tQGLFHV�GH�HVWLPXODomR�REWLGRV�QR�WHVWH�SDUD�R�FURPR�IRUDP�GH���H���
________________________________________________5HYLVmR�GD�OLWHUDWXUD 34
Em 1981, Al-Tawil et al. 169 compararam os resultados do TPL entre
31 pacientes com TC positivo para FURPR� ���� RX� PDLV�� H� ��� LQGLYtGXRV�
VDXGiYHLV�� 2V� DXWRUHV� WHVWDUDP� �� VDLV� GH� FURPR� WULYDOHQWHV� �&U&O3 e
Cr4(SO4)5(OH)2) e dois hexavalentes (K2Cr2O7 e Na2CrO4) em diversas
FRQFHQWUDo}HV� HP� FXOWXUDV� GH� OLQIyFLWRV� FRP� GXUDomR� GH� �� D� �� GLDV�� 9LQWH�
pacientes (sensibilidade = 65%) e 2 controles (especificidade = 92%)
UHVSRQGHUDP�DR�WHVWH��2�DXWRU�HVWDEHOHFHX�FRPR�FULWpULR�SDUD�SRVLWLYLGDGH�D�
SUHVHQoD� GH� ,(� PDLRU� TXH� �� H� VXSHULRU� DR� ,(� GH� XP� FRQWUROH� WHVWDGR�
simultaneamente. Os melhores resultados foram obtidos com as culturas de
��D���GLDV�GH�GXUDomR�FRP�FORUHWR�GH�FURPR�QDV�FRQFHQWUDo}HV�ILQDLV�GH�����
50 e 100 g/ml (P < 0,001). Os pacientes que responderam ao cromo
KH[DYDOHQWH�WDPEpP�UHVSRQGHUDP�DRV�FRPSRVWRV�WULYDOHQWHV��2�DXWRU�WHVWRX�
a especificidaGH� GR� WHVWH� HP� �� SDFLHQWHV� VHQVLELOL]DGRV� DR� QtTXHO� H�RX�
cobalto sendo os resultados negativos.
(P�������5lVlQHQ�et al.27�WHVWDUDP�YiULDV�FRQFHQWUDo}HV�GH�&U&O3,
K2Cr2O7 e Na2CrO4� HP� FXOWXUDV� GH� OLQIyFLWRV� GH� �� D� �� GLDV�� 6HLV� GRV� ��
SDFLHQWHV� DOpUJLFRV� e nenhum dos 8 controles responderam. Os autores
encontraram as melhores respostas nas culturas de 7 e 8 dias com 25 e 50
g/ml de CrCl3 e 0,025; 0,050 e 0,1 g/ml de K2Cr2O7 e Na2CrO4.
(P�VtQWHVH��RV�WUDEDOKRV�XVDUDP�WpFQLFDV�GLIHUHQWHV��GLIHUHQWHV�VDLV
GR� PHWDO�� FRQFHQWUDo}HV� GLYHUVDV�� SHUtRGRV� GH� LQFXEDomR� YDULiYHLV� H� os
FULWpULRV� SDUD� SRVLWLYLGDGH� QmR� foram uniformes. Face aos diferentes
protocolos empregados, Martins et al. 24 compararam os resultados do TPL
entre 20 pacientes com DCA ao cromo e 20 controles utilizando diversos
________________________________________________5HYLVmR�GD�OLWHUDWXUD 35
SDUkPHWURV��)RUDP�UHDOL]DGDV�FXOWXUDV�GH�FXUWD�H�ORQJD�GXUDomR����H���GLDV��
empregando diferentes�FRQFHQWUDo}HV��0,000133 a 133,23 g/mL) de um sal
trivalente (cloreto de cromo) e (0,147 g/mL a 147 g/mL) de outro
hexavalente (biFURPDWR�GH�SRWiVVLR���2V�DXWRUHV�FRQFOXtUDP�TXH�FXOWXUDV�GH�
ORQJD�GXUDomR�HVWLPXODGDV�FRP�333,33 g/mL de cloreto de cromo oferecem
D� PHOKRU� DFXUiFLD� SDUD� R� WHVWH�� Doze casos e apenas um controle
resSRQGHUDP� DR� PpWRGR� XWLOL]DQGR� HVVHV� SDUkPHWURV, determinando uma
sensibilidade de 60% e especificidade de 95%��$SHVDU�GDV�GLIHUHQoDV�QDV�
metodologias empregadas entre os artigos publicados a sensibilidade e
HVSHFLILFLGDGH�GR�PpWRGR�IRL�VHPHOKDQWH�QRV�HVWXGRV�FRP�maior�FDVXtVWLFD��
24,167,169
Os motivos para uma�VHQVLELOLGDGH�DTXpP�GR�GHVHMDGR�QHVVH�WHVWH�
SRGHP� VHU� GHYLGR�� D�� DR� HPSUHJR� GH� PRQyFLWRV� SDUD� D� DSUHVHQWDomR�
DQWLJrQLFD� QR� OXJDU� GDV� FpOXODV� GHQGUtWLFDV� TXH� VmR� PHOKRUHV�
DSUHVHQWDGRUDV�GH�DQWtJHQR��E��EDL[D�VHQVLELOLGDGH�GR�PpWRGR�GH�GHWHFomR�H�
F��DXVrQFLD�GH�XP�JDWLOKR�SDUD�GDU�LQtFLR�D�UHDomR�³LQ�YLWUR´�
5lVlQHQ�et al.27�WHVWDUDP�R�73/�XWLOL]DQGR�FpOXODV�GH�/DQJHUKDQV�HP�
TXDWUR� SDFLHQWHV� H� REWLYHUDP� XP� LQFUHPHQWR� QR� tQGLFH� GH� HVWLPXODomR��
'HQWUH� HVVHV� SDFLHQWHV� VH� HQFRQWUDYD� XP� TXH� QmR� KDYLD� UHVSRQGLGR� Do
WHVWH� FRQYHQFLRQDO�� R� tQGLFH� GH� HVWLPXODomR� VDOWRX� GH� ����� SDUD� ������
tornando-se positivo se considerarmos IE>3 como valor de corte. Outros 3
pacientes, que responderam ao teste convencional, apresentaram aumento
GH�����D�����YH]HV�QR�tQGLFH�GH�HVWLPXODomR�� ,QIHOL]PHQWH�QHQKXP�FRQWUROH�
IRL�WHVWDGR�FRP�HVVH�PpWRGR��p�LPSURYiYHO��SRUpP�SRVVtYHO�TXH�RV�FRQWUROHV�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 36
WDPEpP�DSUHVHQWDVVHP�XP�LQFUHPHQWR�QR�,(��$OpP�GLVVR��R�PpWRGR�p�PXLWR�
WUDEDOKRVR��QmR�SRGHQGR�VHU�XWLOL]DGR�KRMH�SDUD�VXEVWLWXLU�R�WHVWH�GH�FRQWDWo
FRPR�URWLQD�QR�GLDJQyVWLFR�GD�'&$.
Trattner et al. 28 seguiram outro caminho, tentaram aprimorar o
PpWRGR� PHGLQGR por ELISA o INF- no sobrenadante das culturas de 24
horas de����SDFLHQWHV�DOpUJLFRV�DR�FURPR�H����FRQWUROHV��As culturas foram
estimuladDV�FRP�ELFURPDWR�GH�SRWiVVLR������H��� cg/mL) e fitohemaglutinina,
XP� PLWyJHQR� XWLOL]DGR� SDUD� LQLFLDU� D� UHDomR�� Quatorze pacientes e 6
controles responderam, KDYHQGR�GLIHUHQoD�HQWUH�RV�JUXSRV (P = 0,001). Em
FRPSDUDomR� FRP� R� WHVWH� GH� FRQWDWR� D� GHWHFomR� GH� INF- apresentou
sensibilidade e especificidade de 73,7% e 71,4% respectivamente.
Resultados ainda pouco encorajadores.
Em 2005, Moed et al.29 estudaram a resposta proliferativa de 4
pacientes DOpUJLFRV ao cromo em culturas de 6 dias com diferentes
concHQWUDo}HV�GH�&U&O3 (1, 10 e 100 g/ml) e K2Cr2O7 (0,01; 0,1 e 1 g/ml).
Nenhum dos pacientes respondeu. Os autores passaram a estimular as
culturas de 6 dias FRP�DV�FRPELQDo}HV�GH�FLWRFLQDV�,/-7 e IL-12 e IL-7 e IL-4
H�DYDOLDUDP�D�SUROLIHUDomR�H�D�SURGXomR de INF- e IL-5. A IL-7 foi escolhida
SRU�DXPHQWDU�D�DWLYDomR�FHOXODU�H�D�,/-12 e IL-��SRU�HVWLPXODUHP�D�SURGXomR�
de citocinas Th1 (IFN- ) e Th2 (IL-5) respectivamente. (VVH� PpWRGR� IRL�
aplicado em 7 casos e 7 controles. Os resultados variaram de acordo com o
PpWRGR�GH�GHWHFomR�XWLOL]DGR��7UrV�FDVRV�H�QHQKXP�FRQWUROH� UHVSRQGHUDP�
TXDQGR� R� PpWRGR� GH� DIHULomR� IRL� R� tQGLFH� GH� HVWLPXODomR�� �� FDVRV� H� XP�
FRQWUROH�FRP��D�GHWHFomR�GH�,1)- e os sete casos e um controle quando a
________________________________________________5HYLVmR�GD�OLWHUDWXUD 37
DIHULomR�IRL�IHLWD�SHOD�GHWHFomR�GH�,/-5. O mesmo resultado foi obtido com a
GHWHFomR� GH� ,/-�� HP� VHWH� SDFLHQWHV� DOpUJLFRV� DR� QtTXHO� H� VHWH� FRQWUROHV��
PDV� UHVXOWDGRV� LQIHULRUHV� IRUDP� HQFRQWUDGRV� FRP� R� FREDOWR�� IUDJUkQFLD��
parafenilenodiamina e metilcloroisotiazolinona.
Akerman et al.30 reali]DUDP�D�GHWHFomR�VLPXOWkQHD�GH����FLWRFLQDV�
no sobrenadante das culturas de 6 dias de 20 pacientes DOpUJLFRV ao cromo
XVDQGR� R� NLW� ³/XPLQH[� OLTXLFKLS� DUUD\´�� 7RGDV� DV� citocinas testadas (GM-
CSF, IL- -2, IL-4, IL-5, IL-6, IL-8, IL-10, INF- , e TNF-
TNF-
esses resultados podem levar ao desenvolvimento de testes, baseados na
GHWHFomR� GH� FLWRFLQDV�� SDUD� R� GLDJQyVWLFR� GD� KLSHUVHQVLELOLGDGH� DR� FURPR��
,QIHOL]PHQWH�RV�WHVWHV�QmR�IRUDP�UHDOL]DGRV�HP�XP�JUXSR�FRQWUROH�
Minang et al.31� GHWHFWDUDP� D� SURGXomR� GH� ,/-2, IL-4 e IL-13 por
ELISpot e IFN- , IL-2 e IL-13 por ELISA nas culturas de 20 a 40 horas de
FpOXODV�PRQRQXFOHDUHV�GR�VDQJXH�SHULIpULFR�GH����SDFLHQWHV�FRP�HF]HPD�H�
WHVWH� GH� FRQWDWR� SRVLWLYR� SDUD� RV� PHWDLV� QtTXHO�� FREDOWR�� FURPR�� SDOiGLR� H�
RXUR�H�GH���FRQWUROHV�VDXGiYHLV�FRP�WHVWH�GH�FRQWDWR�QHJDWLYR��$� LQWHQomR�
dos autores era determinaU�VH�D�UHVSRVWD�GRV�GHPDLV�PHWDLV�p�XPD�UHVSRVWD�
PLVWD�� WDQWR� 7K�� TXDQWR� 7K��� FRPR� HVWi� EHP� GHWHUPLQDGR� SDUD� R� QtTXHO��
&RQFOXtUDP�TXH�WRGRV�RV�PHWDLV�WHVWDGRV�OHYDP�D�XPD�UHVSRVWD�PLVWD��FRP�
DXPHQWR�QD�SURGXomR�GH�FLWRFLQDV�SDGUmR�7K��H�7K���'HQWUH�RV�SDFLentes,
QRYH� HUDP� DOpUJLFRV� DR� FURPR�� 3DUD� R� (/,6$� RV� DXWRUHV� DGRWDUDP� FRPR�
FULWpULR�GH�UHVSRVWD�DR�FURPR�XP�LQFUHPHQWR�QR�YDORU�HQFRQWUDGR�QD�FXOWXUD�
HVWLPXODGD� VREUH� D� DIHULomR� GD� FXOWXUD� EDVDO� DOLDGR� D� XPD� UD]mR� HQWUH� D�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 38
cultura estimulada e a cultura basal que deveriam ser respectivamente da
ordem de: - ,
IL-2 e -����3DUD�R�(/,6SRW�RV�FULWpULRV�IRUDP�
os mesmos mas o aumento nas citocinas deveria ser de: ³VSRWV´ e 2
vezes para a IL-2, ³VSRWV´ e 2 vezes para a IL-4 e ³VSRWV´ e 2 vezes
para a IL-13. Seis dos nove pacientes apresentaram resposta a pelo menos
XPD�GDV�FLWRFLQDV�HP�SHOR�PHQRV�XP�GRV�PpWRGRV��(/,6$�RX�(/,6SRW��2V�
WUrV�SDFLHQWHV�TXH� QmR� UHVSRQGHUam foram os que apresentaram resposta
de menor intensidade ao cromo no TC, apenas eritema e edema sem a
IRUPDomR� GH� SiSXODV� RX� YHVtFXODV�� $OpP� GLVVR�� RV� WUrV� DSUHVHQWDUDP�
SRVLWLYLGDGH�LQWHQVD��FRP�IRUPDomR�GH�YHVtFXODV��DR�QtTXHO��XP�DR�FORUHWR�GH�
cobalto H� RXWUR� HULWHPD�H� HGHPD� DR� SDOiGLR� H� RXUR�� $VVLP� D� DXVrQFLD� GH�
positividade nesses pacientes pode se dever a baixa sensibilidade do
PpWRGR�RX�D�UHDo}HV�IDOVR-positivas com os metais.
Em 2008, Lindemann et al.32� WHVWDUDP� RV� PpWRGRV� FHOXODUHV� GH�
GHWHFomR� ³LQ�YLWUR´� GH�DOHUJLD�DR�FURPR�D� ILP� GH� HVWDEHOHFHU� D�FDSDFLGDGH�
GRV� PHVPRV� GLVFHUQLUHP� DOHUJLD� GH� VHQVLELOL]DomR�� HVWDGR� HP� TXH� Ki�
UHDWLYLGDGH�QR�WHVWH�GH�FRQWDWR��PDV�DR�FRQWUiULR�GD�DOHUJLD�QmR�DR�FRQWDWR�
KDELWXDO� FRP� R� DJHQWH� HP� TXHVWmR�� /HPEUDP� TXH� UHDo}HV� LUULWDQWHV� VmR�
FRPXQV� QR� 7&� SDUD� R� FURPR�� DWp� XPD� HP� GXDV� UHDo}HV� VmR� LUULWDQWHV�
TXDQGR� R� FURPR� p� WHVWDGR� QD� IRUPD� GH� ELFURPDWR� GH� SRWiVVLR� D� ������
GLILFXOWDQGR� R� GLDJQyVWLFR�170 Assim um TC positivo para o cromo tem
VLJQLILFDGR� FOtQLFR� OLPLWDGR�� GHYHQGR� VHU� FRUUHODFLRQDGR� j� FOtQLFD� GR�
paciente.171�7RGDYLD��QRV�FDVRV�FU{QLFRV�GH�'&��HYHQWR�EDVWDQWH�FRPXP�QD�
________________________________________________5HYLVmR�GD�OLWHUDWXUD 39
DOHUJLD�DR�FURPR��p�GLItFLO�GLIHUHQFLDU�D�'&,�GD�'&$�SHOR�TXDGUR�FOtQLFR�32,163
(VVHV� GDGRV� OHYDUDP� RV� DXWRUHV� D� EXVFDU� SRU� PpWRGRV� GLDJQyVWLFRV�
aOWHUQDWLYRV� TXH� SRVVDP�SUHIHUHQFLDOPHQWH� GLVWLQJXLU� HQWUH�VHQVLELOL]DomR� H�
alergia. Os autores estudaram o TC, TPL e ELISpot em 37 pacientes
DOpJLFRV�DR�FURPR�����VHQVLELOL]DGRV�H����YROXQWiULRV��
���&RP� UHODomR� DR� 73/� GHPRQVWUDUDP� TXH� FXOWXUDV� GH� �� GLDV�
estimuladas com 12,5 g/mL de cloreto de cromo ou 50 ng/mL de bicromato
GH� SRWiVVLR� VmR� FDSD]HV� GH� GLVFHUQLU� HQWUH� DOpUJLFRV� H� VHQVLELOL]DGRV�� D�
SUROLIHUDomR� OLQIRFLWiULD� IRL� HVWLPXODGD� QRV� DOpUJLFRV� PDV� QmR� QRV�
sensibilizados. Todavia, o resultado foi signiILFDWLYR� TXDQGR� R� FULWpULR� GH�
UHVSRVWD�DGRWDGR�HUD�R�YDORU�REWLGR�SHOD�VXEWUDomR�GDTXHOH�HQFRQWUDGR�QDV�
FXOWXUDV�HVWLPXODGDV�PHQRV�R�GDV�FXOWXUDV�QmR�HVWLPXODGDV��PDV�QmR�FRP�R�
YDORU� HQFRQWUDGR� SHOD� UD]mR� HQWUH� DV� FXOWXUDV�� DSHVDU� GRV� DXWRUHV� WHUHP�
encontrDGR� FRUUHODomR� HQWUH� RV� GRLV� FULWpULRV� �r = 0.85 e P < 0,0001). A
positividade nas culturas estimuladas com cloreto de cromo e/ou bicromato
GH� SRWiVVLR� RFRUUHX� HP� ��� GH� ��� SDFLHQWHV� DOpUJLFRV� ������� �� GH� ���
sensibilizados (31%) e 5 de 25 controles (20%) e se mostrou um fator
SUHGLWLYR� IRUWH� SDUD� DOHUJLD�� WDQWR� QD� FRPSDUDomR� FRP� SDFLHQWHV�
sensibilizados quanto controles, P = 0,004 e < 0,0001 respectivamente.
O ELISpot foi utilizado para dosar as citocinas IFN- , IL-2, IL-4,IL-10
e IL-12 nas culturas de 48� KRUDV�GRV� SDFLHQWHV� DOpUJLFRV�� VHQVLELOL]DGRV�H�
FRQWUROHV�� 1mR� KRXYH� GLIHUHQoD� QD� SURGXomR� GHVVDV� FLWRFLQDV� HQWUH� RV�
JUXSRV� GH� SDFLHQWHV� DOpUJLFRV� H� VHQVLELOL]DGRV�� 2V� SDFLHQWHV� DOpUJLFRV�
DSUHVHQWDUDP� PDLRU� SURGXomR� GH� ,)1- que o os controles (P < 0,05). A
________________________________________________5HYLVmR�GD�OLWHUDWXUD 40
resposta do IFN- ��PDV�QmR�D�GDV�GHPDLV�FLWRFLQDV��VH�FRUUHODFLRQRX�D�FRP�
a intensidade da resposta encontrada no TC (r = 0.49 e P = 0.002). Em
FRQWUDVWH��QmR�KRXYH�FRUUHODomR�HQWUH�DV�UHVSRVWDV�HQFRQWUDGD�QR�7&�H�QR�
TPL. Em conjunto, alergia se correlacionou a resposta no TPL e a resposta
do TC a do IFN- �GHWHFWDGR�SHOR�(/,6SRW��2V�DXWRUHV�FRQFOXHP�TXH�R�73/�p�
PHOKRU�TXH� D�GHWHFomR� GH�FLWRFLQDV� SHOR�(/,6SRW�SDUD� GLVWLQJXLU� SDFLHQWHV�
DOpUJLFRV�GH�VHQVLELOL]DGRV�PDV�DGPLWHP�TXH�QRYRV� WUDEDOKRV�TXH�DYDOLHP
GLIHUHQWHV� FRQGLo}HV� GH� FXOWXUD� SDUD� D� GHWHFomR� GH� FLWRFLQDV� GHYHP� VHU�
IHLWRV�SDUD�GHWHUPLQDU�RV�SDUkPHWURV�PDLV�DGHTXDGRV�SDUD�HVVH� WHVWH��(P�
SDUWLFXODU�GHIHQGHP�R�DXPHQWR�QR�SHUtRGR�GH�LQFXEDomR�TXH�FRPR�FKDPDGR�
D�DWHQomR�SRU�.HDWLQJ�et al.172 pode levaU�D�LGHQWLILFDomR�GH�RXWURV�IHQyWLSRV�
GH�FpOXODV�7��FRPR�DV�FpOXODV�7�GH�PHPyULD�FHQWUDO�
���8P� UHVXPR� GRV� SULQFLSDLV� WUDEDOKRV� VREUH� 73/� FRP� FURPR� HVWi�
GLVSRQtYHO�QR�TXDGUR���
________________________________________________5HYLVmR�GD�OLWHUDWXUD 41
Quadro 1 -�7UDEDOKRV�VREUH�WHVWH�GH�SUROLIHUDomR�OLQIRFLWiUia com cromo
Autores &pOXODV Salutilizado
CRQFHQWUDomR�nas culturas*1
3HUtRGR�GH�cultura*1
0pWRGR�GH�DIHULomR
Resultado
Grosfeld165
(1966)/HXFyFLWRV�
totaisK2Cr2O7 0,0294 a
2,94 g/mL 3-4 dias Blastos I
Lischka166
(1971) /HXFyFLWRV�
totais K2Cr2O7 2,94 g/mL 5 dias Blastos S = 87%
E = 80% Christiansen167
(1979) ³3%01�FHOOV´
CrCl3 26,65 g/mL 6 dias IE > 3 S = 62% E = 91%
Al-Tawil169
(1983) ³3%01�FHOOV´
Diversos 25, 50 e 100 g/mL
(CrCl3)
7 e 8 dias IE>1 e > co
S = 65% E = 92%
Carando168
(1985) ³3%01�FHOOV´
CrCl3 0,5 g/mL 56 horas(2,33 dias)
IE S = 100%E = NT
5lVlQHQ27
(1991) ³3%01�FHOOV´
Diversos 25 e 50 g/mL (CrCl3)
7 e 8 dias IE > 2,5 S = 75% E = 100%
Trattner28
(2003) ³3%01�FHOOV´
K2Cr2O7 0,5 e 1 cg/mL 1 dia IFN- S = 74% E = 71%
Moed29
(2005) ³3%01�FHOOV´
CrCl3 e K2Cr2O7
1, 10 e 100 g/mL (CrCl3)
6 dias IE, IFN- e IL-5
S = 100%E = 86%
Martins24
(2006) ³3%01�FHOOV´
CrCl3 333,33 g/mL 6 dias IE > 3 S = 60% E = 95%
Ackerman30
(2006) ³3%01�FHOOV´
K2Cr2O7 ND ND Diversas citocinas*2
P < 0.05*3 NT
Minang31
(2006) ³3%01�FHOOV´
CrCl3 e K2Cr2O7
100 g/mL
(CrCl3)
20 a 40 horas
IL-2, 4, 13 e IFN- *4
S = 67%*5
E = 100%*5
Lindemann32
(2008) ³3%01�FHOOV´
CrCl3 e K2Cr2O7
12,5 g/mL*6
50 g/mL*7
7 e 8 dias*8 48 horas*9
IE, cpm, citocinas*10
S = 70%*11 E = 80%*11
*1 ±� RV� SDUkPHWURV� TXH� OHYDUDP� DRV� PHOKRUHV� UHVXOWDGRV�� .2Cr2O7 ±� ELFURPDWR� GH� SRWiVVLR�� ,� ±inconclusivo; S ± sensibilidade; E ± especificidade; ³3%01�FHOOV´�±�FpOXODV�PRQRQXFOHDUHV�GR�VDQJXH�SHULIpULFR�� CrCl3 ± cloreto de cromo; IE ±� tQGLFH� GH� HVWLPXODomR�� FR� ± controle testado simultaneamente; NT ±�QmR�WHVWDGD�SHOD�DXVrQFLD�GH�JUXSR�FRQWUROH��1'�±�QmR�GLVSRQtYHO��*2 ± GM-CSF, IL- -2, IL-4, IL-5, IL-6, IL-8, IL-10, INF- , e TNF- ; *3 ± aumento de todas as citocinas, exceto TNF- ; *4 ± o valor de corte foi diferente para cada citocina; *5 ± considerando todas as citocinas testadas; *6 ± melhor valor de CrCl3 para disWLQJXLU�SDFLHQWHV�DOpUJLFRV�GH�VHQVLELOL]DGRV��*7 ±valor de CrCl3�FRP�R�TXDO�KRXYH�GLIHUHQoD�QD�SURGXomR�GH�,)1- �HQWUH�DOpUJLFRV�H�QmR�DOpUJLFRV��*8 ±PHOKRU� SHUtRGR� SDUD� R� WHVWH� GH� SUROLIHUDomR� FRQYHQFLRQDO�� *9 ±� SHUtRGR� XVDGR� SDUD� D� GHWHFomR� GH�citocinas; *10 ±�RV�YDORUHV�GH�FRUWH�QmR�IRUDP�GHVFULWRV�H�DV�FLWRFLQDV�WHVWDGDV�IRUDP��,)1- , IL-2, IL-4, IL-10 e IL-12; *11 ±�YDORU�SDUD�R�WHVWH�GH�SUROLIHUDomR��DV�UHVSRVWD�DV�FLWRFLQDV�QmR�IRUDP�GHVFULWDV�
0e72'26
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__________________________________________________________0pWRGRV 42
4 0pWRGRV
4.1 6HOHomR�GH�SDFLHQWHV
37� SDFLHQWHV� IRUDP� VHOHFLRQDGRV� QR� DPEXODWyULR� GH� DOHUJLD� GD�
'LYLVmR�GH�'HUPDWRORJLD�GR�+RVSLWDO�GDV�&OtQLFDV�GD�)DFXOGDGH�GH�0HGLFLQD�
GD� 8QLYHUVLGDGH� GH� 6mR� 3DXOR (HC-FMUSP) entre novembro de 2009 e
janeiro de 2010. Dezoito pacientes apresentavam�KLVWyULD��FOtQLFD e teste de
contato recente, teste realizado a menos de dois anos,� FRPSDWtYHLV� FRP�
DCA por cromo e formaram R�JUXSR�GH�FDVRV��2V�GDGRV�GD�KLVWyULD�OHYDGRV�
HP� FRQVLGHUDomR� IRUDP�� j� H[LVWrQFLD� GH� H[SRVLomR� DR cromo, o tempo
WUDQVFRUULGR� HQWUH� D� H[SRVLomR� H� R� VXUJLPHQWR� GDV� OHV}HV�� D� UHFRUUrQFLD�
IUHQWH�jV�H[SRVLo}HV�UHSHWLGDV�H�D�PHOKRUD��DR�PHQRV�SDUFLDO��QD�DXVrQFLD�
GD� H[SRVLomR�� &RP� UHODomR� DR� H[DPH� ItVLFR� HVSHFLDO� DWHQomR� IRL� GDGD� j�
morfologia, que deveria VHU� FRPSDWtYHO� FRP� HF]HPD� H� D� WRSRJUDILD� GDV�
OHV}HV�� TXH� GHYHULDP� RFRUUHU� QRV� ORFDLV� GD� H[SRVLomR�� 2V� RXWURV� ��
pacientes tinham KLVWyULD�H�FOtQLFD�QmR�FRQGL]HQWHV�FRP�'&$�SRU�FURPR bem
como teste de contato recente negativo para o metal, esses pacientes foram
iQFOXtGRV� QR� JUXSR� GH� FRQWUROHV�� 1HVVH� HVWXGR� IRUDP� H[FOXtGRV� SDFLHQWHV�
menores de 18 anos idade, gestantes ou�TXH�IL]HUDP�XVR�GH�FRUWLFRVWHUyLGH�
VLVWrPLFR�SRU�XP�SHUtRGR�PtQLPR�GH���VHPDQDV�DQWHV�GD�UHDOL]DomR�GR�7&�H�
do TPL.
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__________________________________________________________0pWRGRV 43
Antes da coleta do exame todos os pacientes foram informados
quanto ao procedimento e a finalidade do estudo. Receberam e assinaram o
termo de consentimento livre e esclarecido.
&DUDFWHUtVWLFDV�GRV�JUXSRV�
1) Grupo de casos: 18 pacientes, sendo 17 GR� JrQHUR� PDVFXOLQR� e 1
feminino, com idade entre 36 e 65� DQRV� �PpGLD� 50 anos)
DSUHVHQWDQGR� KLVWyULD� H� FOtQLFD� FRPSDWtYHO� FRP� VHQVLELOL]DomR� DR�
cromo confirmada por teste de contato.
2) Grupo controle: 19 pacientes, sendo 11� GR� JrQHUR� PDVFXOLQR e 7
feminino com idade entre 26 e 68 anos �PpGLD�44�DQRV��VHP�KLVWyULD�
RX� FOtQLFD� FRPSDWtYHO� FRP� VHQVLELOL]DomR� DR� FURPR� FRP� WHVWH� GH�
FRQWDWR�QHJDWLYR�SDUD�ELFURPDWR�GH�SRWiVVLR��
4.2 Teste de contato
���7RGRV� RV� WHVWHV� GH� FRQWDWR� IRUDP� UHDOL]DGRV� QR� DPEXODWyULR� GH�
Dermatologia do HC-FMUSP. Os testes foram efetuados utilizando a bateria
brasileira padronizada de teste de contato (IPI-ASAC Brasil , Rio de Janeiro,
Brasil).� (VVD� EDWHULD� p� IRUPDGD� SRU� ��� VXEVWkQFLDV� VHQGR� R� ELFURPDWR� GH�
SRWiVVLR� WHVWDGR� D� �����HP� YDVHOLQD�VyOLGD��Dois controles tambpP� IRUDP�
WHVWDGRV� FRP� D� EDWHULD� GH� WHVWH� GH� FRQWDWR� SDUD� FRVPpWLFRV� (IPI-ASAC
Brasil , Rio de Janeiro, Brasil)�H�XP�FRP�R�SUySULR�HVPDOWH.�$V�VXEVWkQFLDV�
TXH� FRPS}HP� HVVDV� EDWHULDV� HVWmR� D� GLVSRVLomR� QRV� TXDGURV� �� H� ���
GLVSRQtYHLV�QRV�DQH[RV�
__________________________________________________________0pWRGRV 44
���2V� DOpUJHQRV� IRUDP� DSOLFDGRV� QR� GRUVR�VXSHULRU� FRP�R� DX[tOLR� GH�
contensores (IPI-ASAC Brasil , Rio de Janeiro, Brasil ± registro no MS-
$19,6$�Q�� ��������������) presos por fita adesiva KLSRDOHUJrQLFDV� (3M )
DSyV� SUHSDUR� GD� SHOH� FRP� VROXomR� GH� pWHU�� 2V� SDFLHQWHV� UHFHberam as
RULHQWDo}HV� QHFHVViULDV� H� DSyV� ���KRUDV�R� WHVWH� HUD� UHWLUDGR� H� D� SULPHLUD�
leitura feita. Em 96 horas o paciente retornava para a segunda leitura e
RULHQWDo}HV�TXDQWR�DRV�UHVXOWDGRV�
���2V�FULWpULRV�XWLOL]DGRV�SDUD�DYDOLDU�RV�UHVXOWDGRV�IRUDP�HVWDEelecidos
SHOR� ³International Contact Dermatitis Research Group´�± ICDRG.173 Dessa
forma os resultados foram divididos em:
negativo (-���VHP�UHDomR�
duvidoso (+?): eritema mal definido;
SRVLWLYR�IUDFR������HULWHPD�GHILQLGR�H�SiSXOD�
positivo forte (++): eriWHPD��SiSXOD�H�YHVtFXODs;
SRVLWLYR�PXLWR�IRUWH��������HULWHPD��SiSXOD e�YHVtFXODV�FRQIOXHQWHV�
5,��UHDomR�LUULWDWLYD�
3DUD�VHU�FRQVLGHUDGR�VHQVLELOL]DGR�DR�DOpUJHQR�WHVWDGR�o resultado
deveria ser positivo em 96 horas com intensidade igual ou superior j� GD�
leitura de 48 horas. Nos casos em que houve positividade de 3 ou mais
VXEVWkQFLDV�QmR�UHOacionadas o teste foi repetido.
__________________________________________________________0pWRGRV 45
4.3 Teste de pUROLIHUDomR�lLQIRFLWiULD
��)RUDP�FROKLGRV�TXLQ]H�PLOLOLWURV�GH�VDQJXH�YHQRVR�SHULIpULFR�GH�FDGD�
paciente, em tXER�HVWpULO� KHSDULQL]DGR��QR�DPEXODWyULR�GH�'HUPDWRORJLD�GR�
HC-FMUSP. O sangue foi processado dentro de 2�KRUDV�QR�/DERUDWyULR�GH�
,QYHVWLJDomR� HP� $OHUJLD� H� ,PXQRORJLD� &OtQLFD� H� ([SHULPHQWDO� �/,0� ���� GD�
FMUSP.
Em fluxo laminar, o sangue era transferido pDUD�XP�WXER�F{QLFR�GH�
��PO� DR� TXDO� HUD� DGLFLRQDGR� LJXDO� TXDQWLGDGH� GH�VROXomR� VDOLQD� LVRW{QLFD��
��PO��$SyV�KRPRJHQHL]DU�R�FRQWH~GR�R�VDQJXH�HUD�WUDQVIHULGR�OHQWDPHQWH��
DWUDYpV� GH� XPD� SLSHWD� GH� WUDQVIHUrQFLD�� SDUD� RXWUR� WXER� F{QLFR de 50ml
contendo 12,5ml de Ficoll-Hypaque. O Ficoll-+\SDTXH� p� XP� JUDGLHQWH� GH�
GHQVLGDGH�TXH�IDFLOLWD�D�VHSDUDomR�H�FROKHLWD�GDV�FpOXODV�PRQRQXFOHDUHV�GR�
UHVWDQWH�GR�VDQJXH�DSyV�D�FHQWULIXJDomR��
���$� FHQWULIXJDomR� HUD� IHLWD� D� ����USP� SRU� ��� PLQXWRV� HP� XPD�
FHQWUtIXJD�UHIULJHUDGD�H�R�FRQWH~GR��VDQJXH�H�)LFROO-Hypaque) separava-se
em fases. A superior era formada pelo plasma, amarelo citrino. Logo abaixo,
D� VHJXQGD� IDVH�� IRUPDGD� SHODV� FpOXODV� PRQRQXFOHDUHV�� DSUHVHQWDYD-se
FRPR� XP� DQHO� HVEUDQTXLoDGR�� $� WHUFHLUD� IDVH� HUD� FRPSRVWD� SHOR� )icoll-
+\SDTXH�� LQFRORU� RX� OLJHLUDPHQWH� HVEUDQTXLoDGR� H� D� TXDUWD�� QD� SRUomR�
LQIHULRU�GR�WXER��HUD�IRUPDGD�SHORV�HULWUyFLWRV�H�JUDQXOyFLWRV�TXH�FRQIHULDP�D�
cor vermelha.
__________________________________________________________0pWRGRV 46
���$SHQDV� R� DQHO� GH� FpOXODV� PRQRQXFOHDUHV� HUD� WUDQVIHULGR�� SHOR�
DX[tOLR� GH� XPD� SLSHWD� GH� WUDQVIHUrQFLD�� SDUD� XP� WXER� F{QLFR� GH� ��PO�
FRQWHQGR��PO�GH�VRUR�ILVLROyJLFR��$SyV�D� WUDQVIHUrQFLD�R�FRQWH~GR�GR� WXER�
era completado para 10ml FRP�VRUR�ILVLROyJLFR�H�FHQWULIXJDGR�D�����USP�SRU�
��� PLQXWRV� HP� FHQWUtIXJD� UHIULJHUDGD�� (VVD� HWDSD� YLVD� HOLPLQDU� o Ficoll-
+\SDTXH�UHVLGXDO�GHYLGR�j�VXD�DomR�FLWRWy[LFD�
���$R� WpUPLQR� GD� FHQWULIXJDomR� R� VREUHQDGDQWH� HUD� GHVSUH]DGR� H� R�
ERWmR�GH�FpOXODV�SUHVHQWH�QR� IXQGR�GR� WXER�F{QLFR�HUD�UHVXVSHQVR�HP��PO�
GH� VRUR� ILVLROyJLFR�� 2� YROXPH� HUD� QRYDPHQWH� FRPSOHWDGR� para 10ml e
centrifugado a 1100rpm por 10 minutos em centrifuga refrigerada.
���1RYDPHQWH��R�VREUHQDGDQWH�HUD�GHVSUH]DGR�H�R�ERWmR�UHVXVSHQVR��
SRUpP�QHVVD�HWDSD��HP��PO�GH�530,�������Rosewell Park Memorial Institute
FXOWXUH� PpGLXP). O RPMI 1640 foi o meio de cultura empregado nas
culturas.
���$SyV� D� KRPRJHQHL]DomR� GR� FRQWH~GR� ��� l� GD� VXVSHQVmR� HUDP�
OHYDGRV�SDUD�D�FRQWDJHP�GH�FpOXODV�QR�DSDUHOKR�Cell Dyn 1400��$V�FpOXODV�
HUDP� WUDQVIHULGDV� SDUD� XP� WXER� F{QLFR contendo RPMI 1640 enriquecido
com 10% de soro humano AB.� $� TXDQWLGDGH� GD� VXVSHQVmR� GH� FpOXODV�
transferida era ajustada para que a VXVSHQVmR� ILQDO� WLYHVVHP� �� [� ��6
FpOXODV�Pl.
__________________________________________________________0pWRGRV 47
4.3.1 7HVWH� GH� SUROLIHUDomR� OLQIRFLWiULD� SDUD� GHWHFomR� GD� DWLYDomR�
FHOXODU�SHOD�WD[D�GH�FDSWDomR�GH�WLPLGLQD�UDGLRPDUFDGD
As cultXUDV�IRUDP�IHLWDV�HP�WULSOLFDWDV�HP�SODFDV�GH����RULItFLRV�FRP�
fundo chato (TPP �� 6ZLW]HUODQG�� (XURSH��� (P� FDGD� RULItFLR� IRUDP�
adicionados 100 O� GD� VXVSHQVmR� GH� FpOXODV� H� ��� O� GH� DQWtJHQR� RX�
PLWyJHQR�D�VHU�WHVWDGR�GLOXtGR�HP�530,��8PD�WULSOLFDWD��FKDPDGD�de basal,
era preparada com 100 O�GH�FpOXODV�H���� O�GH�530,�������VHP�DQWtJHQR�RX�
PLWyJHQR�� (VVD� FXOWXUD� p� D� FXOWXUD� QmR� HVWLPXODGD�� XVDGD� FRPR�
GHQRPLQDGRU�QR�FiOFXOR�GR�tQGLFH�GH�HVWLPXODomR�
O�PLWyJHQR� WHVWDGR� IRi o Pokeweed ± 5 l/ml (PWM - Sigma ). Os
PLWyJHQRV�VmR�XWLOL]DGRV�SDUD�YHULILFDU�D�LQWHJULGDGH�GR�VLVWHPD�LPXQROyJLFR�
2� DQWtJHQR� WHVWDGR� IRi o cloreto de cromo hexahidratado (CrCl3.6H2O ±
fornecido pela Merck, Darmstadt, Alemanha, com pureza 96%). O sal
trivalente foi� GLOXtGR� QR� PHLR� 530,� ����� SDUD� IRUPDU� VROXo}HV� a 10-2 M.
(VVDV� VROXo}HV� IRUDP� KRPRJHQHL]DGDV e estocadas a ±70o&� DWp� R� XVR��
Durante o TPL D� VROXomR� GH� FORUHWR� GH� FURPR� D� ��-2� 0� HUD� GLOXtGD� SDUD�
2,5.10-3 M, equivalente a 666,66 g/mL. $�FRQFHQWUDomR�ILQDO�GDV�FXOWXUDV�HUD�
a metade dessD�FRQFHQWUDomR,�Mi�TXH�DV�FXOWXUDV�HUDP�FRPSRVWDV�SRU���� l
GD�VXVSHQVmR�GH�FpOXODV�H���� O�GD�VROXomR�
���$V� FpOXODV� IRUDP� FXOWLYDGDV� HP� WULSOLFDWDV� SRU� �� GLDV� D� ��oC em
atmosfera de 5% de CO2. O� PLWyJHQR� PWM era testado como controle
positivo.
__________________________________________________________0pWRGRV 48
���'H]RLWR�KRUDV�DQWHV�GR�WpUPLQR�GDV�FXOWXUDV���� l de timidina tritiada
(3H-thymidine, Amershan Pharmacia Biotech, UK; specific activity
25Ci/mmol�� GLOXtGD� HP�530,� ����� �������� IRL� DGLFLRQDGD� HP� FDGD� FXOWXUD�
para ser incorporada SHODV�FpOXODV em quantidade diretamente proporcional
DR�VHX�HVWDGR�GH�DWLYDomR.
���&RPSOHWDGR�R�SHUtRGR�GH�LQFXEDomR�DV�FpOXODV�HUDP�DVSLUDGDV�SRU�
XP�FROHWRU�GH�FpOXODV�DXWRPiWLFR��Cell-Harvester, Skatron Instruments, UK) e
passadas para uma membrana de fibra de vidro que era deixada para secar
por 24 horas.
���$SyV� R�SHUtRGR� GH� VHFDJHP�� DV�PHPEUDQDV� HUDP� EDQKDGDV� FRP�
OtTXLGR� GH� FLQWLODomR� H� VHODGDV� HP� HQYHORSHV� SOiVWLFRV�� 3DVVDYDP� SRU� XP�
FRQWDGRU� GH� FLQWLODomR� �%HWDSODWH�� :DOODF�� )LQOkQGLD) que determinava a
FDSWDomR�GH� WLPLGLQD�Uadiomarcada em cada cultura e dava o resultado em
FRQWDJHQV� SRU� PLQXWR� �FSP��� $� PpGLD� GDV� WULSOLFDWDV� HUD� FDOFXODGD� H� RV�
UHVXOWDGRV�H[SUHVVRV�HP�tQGLFH�GH�HVWLPXODomR�(IE) TXH�p�D� UD]mR�HQWUH�D�
PpGLD�GH� FDSWDomR�GH� WLPLGLQD� UDGLRPDUFDGD� GDV�FXOWXUDV� HVWLPXladas e a
PpGLD�HQFRQWUDGD�QDV�FXOWXUDV�QmR�HVWLPXODGDV�
ËQGLFH�GH�HVWLPXODomR (IE) = _PpGLD�GD�FDSWDomR�QDV�FXOWXUDV�HVWLPXODGDV__ �PpGLD�GD�FDSWDomR�QDV�FXOWXUDV�QmR�HVWLPXODGDV
__________________________________________________________0pWRGRV 49
4.3.2 7HVWH� GH� SUROLIHUDomR� OLQIRFLWiULD� SDUD� GHWHFomR da DWLYDomR�
FHOXODU�SHOD�SURGXomR de citocinas
As culturas foram realizadas em monoplicatas em placas de 96
RULItFLRV� FRP� IXQGR� FKDWR� �733 �� 6ZLW]HUODQG�� (XURSH��� (P� FDGD� RULItFLR�
foram adicionados 100 O�GD�VXVSHQVmR�GH�FpOXODV�H���� l de meio RPMI ou
da� VROXomR� GH� FORUHWR� GH� FURPR� a 2,5.10-3� 0�� IRUPDQGR� FXOWXUDV� QmR�
HVWLPXODGDV�� WDPEpP� GHQRPLQDGDV� GH� � FXOWXUDV� EDVDLV� H� HVWLPXODGDV�
UHVSHFWLYDPHQWH��$V�FXOWXUDV�SDUD�D�GHWHFomR�GH�FLWRFLQDV�IRUDP�SUHSDUDGDV�
DR�PHVPR� WHPSR�H�QDV�PHVPDV�FRQGLo}HV�TXH�DV�FXOWXUDV�SDUD�GHWHFomR�
por timidina radiomarcada, mas em placas diferentes.
���$R� WpUPLQR� GR� SHUtRGR� GH� LQFXEDomR� DV� FXOWXUDV� IRUDP�
FHQWULIXJDGDV�� SDUD� VHSDUDU� DV�FpOXODV� GR� VREUHQDGDQWH� H� R�VREUHQDGDQWH�
WUDQVIHULGR�SDUD� WXERV�SOiVWLFRV� LGHQWLILFDGRV�H�DUPD]HQDdos a -70o&�DWp�D�
dosagem das citocinas.
Os resultados da GHWHFomR�GH�FLWRFLQDV�IRUDP�H[SUHVVRV�HP�SJ�P/��
2V� FULWpULRV� XVDGRV� SDUD� SRVLWLYLGDGH� GRV� WHVWHV� IRUDP�� R� DXPHQWR� QD�
SURGXomR� GH� FLWRFLQDV� GHWHUPLQDGR� SHOD� GLIHUHQoD� HQWUH� D� SURGXomR�
encontrada nDV� FXOWXUDV� HVWLPXODGDV� PHQRV� D� SURGXomR� GD� FXOWXUD� EDVDO��
GLIHUHQoD� HVVD� DTXL� GHQRPLQDGD� GH� GHOWD� � ) e pelD� UD]mR� HQWUH� o valor
encontrado nas culturas estimuladas e as culturas basais, denominada de
tQGLFH�GH�HVWLPXODomR��,(��
Delta ( ) = valor encontrado nas culturas estimuladas valor encontrado nas culturas basais
__________________________________________________________0pWRGRV 50
ËQGLFH�GH�HVWLPXODomR�(IE) = valor encontrado nas culturas estimuladas valor encontrado nas culturas basais
4.4 Dosagem das citocinas no sobrenadante das culturas
4.4.1 ³/XPLQH[�OLTXLFKLS�DUUD\´
���$�GRVDJHP�GDV�FLWRFLQDV�IRL�UHDOL]DGD�XWLOL]DQGR�R�VLVWHPD�³OXPLQH[�
OLTXLFKLS� DUUD\´�� (VVH� VLVWHPD� SHUPLWH� D� GRVDJHP� VLPXOWkQHD� GH� P~OWLSODV�
citocinas no sobrenadante de culturas. Nesse PpWRGR� PLFURHVIHUDV� GH�
poliestireno VmR�FRUDGDV�com dois fluorocromos espectrais distintos (figura
2). 8WLOL]DQGR�XPD�SURSRUomR�SUHFLVD�GHVWHV�GRLV�fluorocromos foram criadas
100 microesferas diferenciadas pela sua cor. &DGD�HVIHUD�p�FRQMXJDGD�D�XP�
anticorpo GH� FDSWXUD� HVSHFtILFR� TXH� VH� OLJDUi� DR� VHX� DQWtJHQR� TXDQGR�
encontra-OR�� $� YDQWDJHP� GHVVH�PpWRGR� UHVLGH� QD� XWLOL]DomR� GH� HVIHUDV� GH�
HVSHFLILFLGDGHV� GLIHUHQWHV�SDUD� D�GHWHFomR�VLPXOWkQHD� GH�YiULRV�DQWtJHQRV�
em uma amostra.
Fig 2. Microesferas de poliestireno coradas de forma singular.
__________________________________________________________0pWRGRV 51
As microsferas foram incubadas com o sobrenadante das culturas.
$SyV� D� LQFXEDomR� D� DPRVWUD� foi lavada para remover as microesferas que
QmR�VH�OLJDUDP�DRs seus respectivos�DQWtJHQRs. Em seguida um anticorpo de
GHWHFomR� ELRWLQLODGR,� TXH� VH� OLJD� DR� FRPSOH[R� DQWtJHQR-anticorpo preso a
microesfera de poliestireno, foi incubado com a amostra. O resultado final
era� DPSOLILFDGR� DWUDYpV� Ga� LQFXEDomR� FRP� R� FRQMXJDGR� HVWUHSWDYLGLQD-
ficoeritrina que emite um sinal fluorescente (figura 3). As microesferas foram
lidas no equipamento Luminex 100 �DWUDYpV�GH�um sistema duplo de lasers
que incide nDV�PLFURHVIHUDV� j�PHGLGD� TXH�HVWDV�passavam pelo aparelho.
Um feixe de laser detectava a cor da microesfera e portanto o ensaio
relacionado a sua cor e o outro laser quantificava o sinal em cada
microesfera (figura 4).
)LJ����$�HVIHUD�FRP�R�DQWLFRUSR�p�LQFXEDGD�FRP�D�DPRVWUD�H�VH�OLJD�DR�VHX�DQWtJHQR��$�DPRVWUD�p� ODYDGD�H�LQFXEDGD�FRP�XP�DQWLFRUSR�ELRWLQLODGR�TXH�se ligD� DR� FRPSOH[R� DQWtJHQR-anticorpo. O resultado p amplificado pela
__________________________________________________________0pWRGRV 52
LQFXEDomR� FRP� R� FRQMXJDGR� HVWUHSWDYLGLQD-ficoeritrina que emite um sinal fluorescente.
)LJ����2V�IHL[HV�GH�ODVHU�LQFLGHP�VREUH�DV�HVIHUDV�SHUPLWLQGR�D�GHWHFomR�GR�ensaio e da intensidade da resposta.
4.4.2 Citocinas testadas
As citocinas escolhidas para estudo foram o IFN- , IL-2, IL-4, IL-5,
IL-10, IL-12, IL-13, IL-��� H� 5$17(6�� 2V� YDORUHV� PpGLRV� PtQLPRV� GH�
GHWHFomR�GHVVDV�FLWRFLQDV�FRP�R�PpWRGR�VmR����������������������������������4;
0,2 e 1,0 J�P/� UHVSHFWLYDPHQWH�� 3DUD� RV� FiOFXORV� HVWDWtVWLFRV� WRGRV� RV�
YDORUHV� HQFRQWUDGRV� DEDL[R� GD� FDSDFLGDGH� GH� GHWHFomR� GR� PpWRGR� IRUDP�
WURFDGRV�SDUD�R�YDORU�PpGLR�PtQLPR�GH�GHWHFomR�GH�FDGD�FLWRFLQD�
__________________________________________________________0pWRGRV 53
4.4 0pWRGRV�HVWDWtVWLFRV
�3DUD� D� DYDOLDomR� HVWDWtVWLFD� IRUDP utilizados os testes� QmR�
SDUDPpWULFRs de Mann-Whitney U H� R� WHVWH� GH� FRUUHODomR� GH� 6SHDUPDQ�
(software GraphPad Prism �YHUVmR������H�RV�UHVXOWDGRV�IRUDP�FRQVLGHUDGRV�
significantes quando P <0,05. A sensibilidade, especificidade�H�DFXUiFLa dos
PpWRGRV foram calculadas.
RESULTADOS
id23705453 pdfMachine by Broadgun Software - a great PDF writer! - a great PDF creator! - http://www.pdfmachine.com http://www.broadgun.com
________________________________________________________Resultados 54
5 RESULTADOS
5.1 6HOHomR�GRV�JUXSRV
O grupo de casos foi formado por 18 pacientes, sendo 17 GR�JrQHUR�
masculino e 1 feminino e o grupo controle por 19 pacientes, 11� GR�JrQHUR�
masculino e 7 feminino. A idade do grupo de casos variou entre 36 e 65
anoV��PpGLD�50 anos) e do grupo controle de 26 a 68�DQRV��PpGLD�44 anos),
QmR�KDYHQGR�GLIHUHQoD�HVWDWtVWLFD�VLJQLILFDWLYD��P = 0.1913).
5.2 Testes de contato
Todos os casos apresentaram teste de contato positivo (+, ++ ou
+++) para cromo. Em um paciente o teste foi positivo fraco (+), em treze
positivo forte (++) e em quatro�SRVLWLYR�PXLWR�IRUWH��������$OpP�GR�FURPR��oito
pacientes eram sensibilizados ao carba-mix, cinco ao tiuram-mix e ao sulfato
GH� QtTXHO, WUrV� DR� FREDOWR� H� XP�ao PPD-PL[�� IRUPDOGHtGR� nitrofurasona e
quinolina.
Todos os controles apresentaram teste de contato negativo para
ELFURPDWR� GH� SRWiVVLR��Cinco pacientes eram sensibilizados ao sulfato de
QtTXHO, dois DR� FREDOWR�� FRORI{QLR�� QHRPLFLQD� H� SHUIXPH-PL[� H� XP� j�
id23710609 pdfMachine by Broadgun Software - a great PDF writer! - a great PDF creator! - http://www.pdfmachine.com http://www.broadgun.com
________________________________________________________Resultados 55
EHQ]RFDtQD��FORUH[LGLQH��HWLOHQRGLDPLQD��HVPDOWH��IRUPDOGHtGR��QLWURIXUD]RQD��
WLPHURVDO��WLRJOLFRODWR�GH�DP{QLD�H�WLXUDP-mix.
2V� UHVXOWDGRV� GRV� WHVWHV� GH� FRQWDWR� GRV� FDVRV� H� FRQWUROHV� HVWmR�
GLVSRQtYHLV�no quadro ���SUHVHQWH�QD�VHVVmR�GH�DQH[RV�
5.3 Testes de pUROLIHUDomR lLQIRFLWiULD
���7DQWR�FRQWUROHV�TXDQWR�FDVRV�DSUHVHQWDUDP�SUROLIHUDomR�DGHTXDGD�
DR� VHUHP� HVWLPXODGRV� FRP� R� PLWyJHQR� pokeweed, P < 0.0001 nos dois
JUXSRV�� QmR� H[LVWLQGR� GLIHUHQoD� QRV� tQGLFHV� GH� HVWLPXODomR� HQWUH� FDVRV� H�
controles, P = 0.2185 (Figuras 5 a 7).
1R�HQWDQWR��DSHQDV�R�JUXSR�GH�FDVRV� UHVSRQGHX�DR�HVWtPXOR�FRP�
cromo, P < 0.0001 e 0.2202 para casos e controles respectivamente (figuras
�� H� ���� 2V� tQGLFHV� GH� HVWLPXODomR� HQFRQWUDGRV� QRV� JUXSRV� GH� FDVRV� H�
FRQWUROHV� HVWLPXODGRV� FRP� FURPR� WDPEpP� IRUDP diferentes, P = 0.0014
(figura 10).
���6HLV� GRV� ��� FDVRV� H� QHQKXP� FRQWUROH� DSUHVHQWDUDP� tQGLFH� GH�
HVWLPXODomR� VXSHULRU� D� ��� DSUHVHQWDQGR� R� WHVWH� GH� SUROLIHUDomR� OLQIRFLWiULD�
uma sensibilidade, especificidade� H� DFXUiFLD� de 33%, 100% e 68%,
respectivamente. Os valores encontrados nas culturas basais e estimuladas
FRP�FURPR��EHP�FRPR�RV�tQGLFHV�GH�HVWLPXODomR�H�RV�YDORUHV�GHOWD�REWLGRV��
HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 56
Fig 5. Contagens por minutoencontradas nas culturas
basais e estimuladas com PWMdo grupo de casos.
Basal casos PWM casos0
1000
2000
3000
4000
P < 0.0001
Fig 6. Contagens por minutoencontradas nas culturas
basais e estimuladas com PWMdo grupo controle.
Basal controles PWM controles0
2500
5000
7500
10000
P < 0.0001
)LJ����ËQGLFHV�GH�HVWLPXODomRencontrados nos casos e
controles estimulados comPWM.
IE PWM casos IE PWM controls0
10
20
30
P = 0.2185
Fig 8. Contagens por minutoencontradas nas culturasbasais e estimuladas comcromo do grupo de casos.
Basal casos Cromo casos0
250
500
750
P < 0.0001
Fig 9. Contagens por minutoencontradas nas culturasbasais e estimuladas comcromo do grupo controle.
Basal controles Cromo controles0
500
1000
1500
P = 0.2202
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nos casos e
controles estimulados comcromo.
IE cromo controles IE cromo casos0
1
2
3
4
5
P = 0.0014
________________________________________________________Resultados 57
5.4 'HWHFomR�GDV�FLWRFLQDV
5.4.1 Interferon-gama
���1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,1)- entre as culturas
basais de casos e controles, P = 0.1406 (figuras 11).
���+RXYH�DXPHQWR�GD�SURGXomR�GH� ,)1- nas culturas estimuladas no
JUXSR� GH� FDVRV�� PDV� QmR� QR� JUXSR� FRQWUROH� - P = 0.0005 e 0.1290
respectivamente (figuras 12 e 13).
���2V�tQGLFHV�REWLGRV�SHOD�UD]mR�HQWUH�D�SURGXomR�GH�,)1- encontrada
QDV� FXOWXUDV� HVWLPXODGDV� FRP� FURPR� H� QDV� QmR� HVWLPXODGDV� QR� JUXSR� GH�
casos foram superiores aos vistos no grupo controle, P = 0.0186 (figura 14).
Da mesma forma os vDORUHV� HQFRQWUDGRV� SHOD� GLIHUHQoD� QD�
SURGXomR� GH� ,)1- � HQWUH� DV� FXOWXUDV� HVWLPXODGDV� H� QmR� HVWLPXODGDV� FRP�
cromo no grupo de casos foram maiores do que os obtidos no grupo
controle, P = 0.0186 (figura 15).
���2V� YDORUHV� GRV� tQGLFHV� GLDJQyVWLFRV� TXH� DSUHVHQtaram a melhor
DFXUiFLD� IRUDP�� ,(� VXSHULRU� D� �� H� maior que 50. Com esses valores de
corte, a sensibilidade e especificidade do teste com o IE foi de 83% e 63%
�DFXUiFLD�GH������H�FRP�R� �GH�����H������DFXUiFLD�GH�������7RGRV�RV�
valores de INF- detectadoV� H� GRV� tQGLFHV� XWLOL]DGRV� HVWmR� GLVSRQtYHLV� QD�
tabela 2, presente nos anexos.
________________________________________________________Resultados 58
)LJ�����'HWHFomR�GH�,)1� nasculturas basais dos casos e
controles.
Basal casos Basal controles0
100
200
300
400
500
P = 0.1406
Fig 12. 'HWHFomR�GH�,)1� nasculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
2500
5000
7500
P = 0.0005
Fig 13. 'HWHFomR�GH�,)1� nasculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
500
1000
1500
P = 0.1290
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
100
200
300
400
500
P = 0.0186
)LJ�����'LIHUHQoD�QD�GHWHFomRde IFN- entre as culturasestimuladas e basais dos
grupos de caso e controle.
casos controles-2500
0
2500
5000
7500
P = 0.0186
________________________________________________________Resultados 59
5.4.2 Interlucina-2
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-2 entre as culturas
basais de casos e controles e nem entre as culturas basais e estimuladas do
grupos, P = 1, 0.2110 e 0.1567, respectivamente (figuras 16 a 18).
���2V� YDORUHV� REWLGRV� SHOD� UD]mR� HQWUH� DV� FXOWXUDV� HVWLPXODGDV� FRP�
cromo e as basais no grupo de casos foram superiores aos vistos no grupo
controle, P = 0.0465 (figura 19).
No entanto, os valorHV�HQFRQWUDGRV�SHOD�GLIHUHQoD�HQWUH�DV�FXOWXUDV�
HVWLPXODGDV� H� QmR� HVWLPXODGDV� QR� JUXSR� GH� FDVRV� QmR� IRUDP� GLIHUHQWHV�
daqueles encontrados no grupo controle, P = 0.0705 (figura 20).
���2V�YDORUHV�yWLPRV�HQFRQWUDGRV� IRUDP�FRP�XP� ,(�VXSHULRU�D������$�
sensibilidade e especificidade do teste com esse IE foram de 61% e 84%
UHVSHFWLYDPHQWH�� DFXUiFLD� GH������7RGRV� RV�YDORUHV�GH� ,/-2 detectados e
GRV�tQGLFHV�XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 60
Fig 16. 'HWHFomR�GH�,L-2 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
5
10
15
P = 1
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
10
20
P = 0.2110
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
5
10
15
P = 0.1567
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
10
20
30
40
P = 0.0465
Fig 20. 'LIHUHQoD�QD�GHWHFomRde IL-2 entre as culturas deestimuladas e basais dos
grupos de caso e controle.
-20
-10
0
10
20
casos controles
P = 0.0705
________________________________________________________Resultados 61
5.4.3 Interleucina-4
Foi encontrado maior pURGXomR�GH�,/-4 nas culturas basais do grupo
GH�FDVRV�HP�UHODomR�DV�FXOWXUDV�EDVDLV�GR�JUXSR�FRQWUROH��P = 0.0164 (figura
21).
���1mR� KRXYH� GLIHUHQoD� HQWUH� RV� YDORUHV� GH� ,/-4 detectados nas
culturas basais e estimuladas no grupo de casos, mas houve aumento nas
FXOWXUDV�HVWLPXODGDV�HP�UHODomR�DV�EDVDLV�QR�JUXSR�FRQWUROH�± P = 0.2114 e
0.0265 respectivamente (figuras 22 e 23).
���2V�YDORUHV�REWLGRV� WDQWR�SHOD�UD]mR�TXDQWR�SHOD�GLIHUHQoD�HQWUH�DV�
FXOWXUDV�HVWLPXODGDV�FRP�FURPR�H�DV�EDVDLV�QR�JUXSR�GH�FDVRV�QmR foram
diferentes daqueles encontrados no grupo controle, P = 0.6268 e 0.7962
respectivamente (figuras 24 e 25). Todos os valores de IL-4 detectados e
GRV�tQGLFHV�XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 62
Fig 21. 'HWHFomR�GH IL-4 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
25
50
75
100
P = 0.0164
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
25
50
75
100
P = 0.2114
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
50
100
150
P = 0.0265
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
1
2
P = 0.6268
Fig 25. 'LIHUHQoD�QD�GHWHFomRde IL-4 entre as culturas
estimuladas e basais dosgrupos de caso e controle.
casos controles-25
0
25
50
P =0.7962
________________________________________________________Resultados 63
5.4.4 Interleucina-5
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-5 entre as culturas
basais dos casos e controles, P = 0.069 (figuras 26).
���+RXYH� DXPHQWR� GD� SURGXomR� GH� ,/-5 nas culturas estimuladas no
JUXSR� GH� FDVRV�� PDV� QmR� QR� JUXSR� FRQWUROH� ± P = 0.0006 e 0.7588
respectivamente (figuras 27 e 28).
���2V�YDORUHV�REWLGRV� WDQWR�SHOD�UD]mR�TXDQWR�SHOD�GLIHUHQoD�HQWUH�DV�
culturas estimuladas e as basais no grupo de casos foram superiores aos
vistos no grupo controle, P = 0.0032 e 0.0102 respectivamente (figuras 29 e
30).
Os valRUHV�yWLPRV�GH�FRUWH�IRUDP��,(�VXSHULRU���H� maior que 0,2. A
VHQVLELOLGDGH�� HVSHFLILFLGDGH� H� DFXUiFLD� GR� WHVWH� FRP� HVVHV� SDUkPHWURV�
foram de 72%, 79% e 76% com esse IE e 56%, 89% e 73% com o . Todos
os valores de IL-��GHWHFWDGRV�H�GRV�tQGLFHV�XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�
tabela 5, presente nos anexos.
________________________________________________________Resultados 64
Fig 26. 'HWHFomR�GH IL-5 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
5
10
15
P = 0.0609
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0.0
2.5
5.0
7.5
10.0
P = 0.0006
)LJ�����'HWHFomR�GH�,/���QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
5
10
15
P = 0.7588
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
25
50
75
100
P = 0.0032
Fig 30. 'LIHUHQoD�QD�GHWHFomRde IL-5 entre as culturas
estimuladas e basais dosgrupos de caso e controle.
-10
0
10
casos controles
P = 0.0102
________________________________________________________Resultados 65
5.4.5 Interleucina-10
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-10 entre as culturas
basais dos casos e controles, P = 0.6595 (figuras 31).
���7RGDYLD�� KRXYH� UHGXomR� QRV� YDORUHV� GHWHFWDGRV� QDV� FXlturas
estimuladas com cromo tanto no grupo de casos quanto no grupo controle, P
< 0.0001 e 0.0001 respectivamente (figuras 32 e 33).
���2V� YDORUHV� REWLGRV� SHOD� UD]mR� H� SHOD� GLIHUHQoD� HQWUH� DV� FXOWXUDV�
HVWLPXODGDV�FRP�FURPR�H�DV�EDVDLV�QR�JUXSR�GH�FDVRV�Qmo foram diferentes
daqueles obtidos no grupo controle, P = 0.2019 e 0.7962 respectivamente
(figuras 34 e 35). Todos os valores de IL-��� GHWHFWDGRV� H� GRV� tQGLFHV�
XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 66
Fig 31. 'HWHFomR�GH IL-10 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
1000
2000
3000
P = 0.6595
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
1000
2000
P < 0.0001
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
1000
2000
3000
P = 0.0001
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0.00
0.25
0.50
0.75
1.00
P = 0.2019
Fig 35. 'LIHUHQoD�QD�GHWHFomRde IL-10 entre as culturasestimuladas e basais dos
grupos de caso e controle.
casos controles-3000
-2000
-1000
0
P = 0.7962
________________________________________________________Resultados 67
5.4.6 Interleucina-12
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-12 entre as culturas
basais de casos e controles, P = 0.0946 (figura 36).
���7DPEpP� QmR� KRXYH� GLIHUHQoD� HQWUH� RV� YDORUHV� GHWHFWDGRV� QDV�
FXOWXUDV� HVWLPXODGDV� H� QmR� HVWLPXODGDV� QRV� GRLV� JUXSRV�� P = 0.1940 e
0.2999 respectivamente (figuras 37 e 38).
���1R� HQWDQWR�� RV� YDORUHV� REWLGRV� WDQWR� SHOD� UD]mR� TXDQWR� SHOD�
GLIHUHQoD�HQWUH�DV�FXOWXUDV�HVWLPXODGDV�H�DV�EDVDLV�QR�JUXSR�GH�FDVRV�IRUDP�
superiores aos vistos no grupo controle, P = 0.0024 e 0.0050 (figuras 39 e
40).
Os melhores valores encontrados para o IE e o foram: superior a
1,5 e 0,3 respectivamente. Com esses, a sensibilidade, especificidade e
DFXUiFLD� GR� WHVWH� IRUDP�GH������ ����H� ����SDUD�R� ,(�H� GH� ����� ����H�
70% para o . Todos os valores de IL-12 detectados e GRV�tQGLFHV�XWLOL]DGRV�
HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 68
Fig 36. 'HWHFomR�GH IL-12 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
10
20
30
P = 0.0946
Fig 37. 'HWHFomR�GH IL-12 nasculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
10
20
P = 0.1940
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
10
20
30
P = 0.2999
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0.0
2.5
5.0
7.5
P = 0.0024
Fig 40. 'LIHUHQoD�QD�GHWHFomRde IL-12 entre as culturasestimuladas e basais dos
grupos de caso e controle.
-30
-20
-10
0
10
20
30
casos controles
P = 0.0050
________________________________________________________Resultados 69
5.4.7 Interleucina-13
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-13 entre as culturas
basais de casos e controles, P = 0.0945 (figuras 41).
���2� FURPR�HVWLPXORX� D� SURGXomR�de IL-13 nas culturas do grupo de
FDVRV�� PDV� QmR� QDV� FXOWXUDV� GR� JUXSR� FRQWUROH�� P < 0.0001 e 0.9186
respectivamente (figuras 42 e 43).
���2V�YDORUHV�HQFRQWUDGRV�WDQWR�SHOD�UD]mR�TXDQWR�SHOD�GLIHUHQoD�HQWUH�
as culturas estimuladas com cromo e as basais no grupo de casos foram
superiores aos obtidos no grupo controle, P� �� �������QDV� GXDV�FRQGLo}HV�
(figuras 44 e 45).
Utilizando os valores superiores a 3 para o IE e superior a 2 para o
��D�VHQVLELOLGDGH��HVSHFLILFLGDGH�H�DFXUiFLD�GR�WHVWH�IRUDP�GH��������% e
89% para o IE e 94%, 89% e 92% para o . Todos os valores de IL-13
GHWHFWDGRV�H�GRV� tQGLFHV�XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD� WDEHOD����SUHVHQWH�
nos anexos.
________________________________________________________Resultados 70
Fig 41. 'HWHFomR�GH IL-13 nasculturas basais dos casos e
controles.
Basal casos Basal controles0
100
200
300
P = 0.0945
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0
25
50
75
P < 0.0001
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0
100
200
300
P < 0.9186
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
100
200
P < 0.0001
Fig 45. 'LIHUHQoD�QD�GHWHFomRde IL-13 entre as culturasestimuladas e basais dos
grupos de caso e controle.
-100
-50
0
50
100
casos controles
P < 0.0001
________________________________________________________Resultados 71
5.4.8 Interleucina-17
1mR�IRL�HQFRQWUDGR�GLIHUHQoD�QD�SURGXomR�GH�,/-17 entre as culturas
basais dos grupos, P = 0.4459 (figura 46).
���7DPEpP�QmR�KRXYH�GLIHUHQoD�QRV�YDORUHV�GHWHFWDGRV�QDV�FXOWXUDV�
HVWLPXODGDV�H�QmR�HVWLPXODGDV�QR�JUXSR�GH�FDVRV�H�QR�JUXSR�FRQWUROH��P =
0.3101 e 0.1547 respectivamente (figuras 47 e 48).
Os valores obtidos pela rD]mR� H� SHOD� GLIHUHQoD� HQWUH� DV� FXOWXUDV�
HVWLPXODGDV�H�DV�QmR�HVWLPXODGDV�QR�JUXSR�GH�FDVRV�QmR� IRUDP�GLIHUHQWHV�
daqueles obtidos no grupo controle, P = 0.0698 e 0.0910 respectivamente
(figuras 49 e 50). Todos os valores de IL-��� GHWHFWDGRV� H� GRV� tQGLFHV�
utLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�WDEHOD����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 72
Fig 46. 'HWHFomR�GH IL-17 nasculturas basais dos casos e
controles.
Basal casos Basal controles0.0
2.5
5.0
7.5
10.0
P = 0.4459
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladas
com cromo do grupo de casos.
Basal casos Cromo casos0.0
2.5
5.0
7.5
P = 0.3101
)LJ�����'HWHFomR�GH�,/����QDVculturas basais e estimuladascom cromo do grupo controle.
Basal controles Cromo controles0.0
2.5
5.0
7.5
10.0
P = 0.1547
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
10
20
30
P = 0.0698
Fig 50. 'LIHUHQoD�QD�GHWHFomRde IL-17 entre as culturasestimuladas e basais dos
grupos de caso e controle.
-10
-5
0
5
10
casos controles
P = 0.0910
________________________________________________________Resultados 73
5.4.9 RANTES
���1mR� IRL� HQFRQWUDGR� GLIHUHQoD� QD� SURGXomR� GH� RANTES entre as
culturas basais de casos e controles, P� ���������ILJXUD������-i�DV�FXOWXUDV�
de casos e controles realizadas com cromo aXPHQWDUDP� D� SURGXomR� GH�
RANTES, P < 0,0001 (figuras 52 e 53).
���-i�RV�YDORUHV�REWLGRV�SHOD�UD]mR�H�SHOD�GLIHUHQoD�HQWUH�DV�FXOWXUDV�
estimuladas com cromo e as basais no grupo de casos foram iguais aos
encontrados no grupo controle, P = 0.5535 e 0.6595 respectivamente
�ILJXUDV���� H� �����7RGRV� RV�YDORUHV�GH�5$17(6�GHWHFWDGRV�H� GRV� tQGLFHV�
XWLOL]DGRV�HVWmR�GLVSRQtYHLV�QD�WDEHOD�����SUHVHQWH�QRV�DQH[RV�
________________________________________________________Resultados 74
)LJ�����'HWHFomR�GH�5$17(6nas culturas basais dos casos
e controles.
Basal casos Basal controles0
2500
5000
7500
P = 0.6161
)LJ�����'HWHFomR�GH�5$17(6nas culturas basais e
estimuladas com cromo dogrupo de casos.
Basal casos Cromo casos0
5000
10000
P < 0.0001
)LJ�����'HWHFomR�GH�5$17(6nas culturas basais e
estimuladas com cromo dogrupo controle.
Basal controles Cromo controles0
2500
5000
7500
P < 0.0001
)LJ�����ËQGLFHV�GH�HVWLPXODomRencontrados nas culturas dos
casos e controles.
IE casos IE controles0
5
10
15
P = 0.5535
Fig 55. 'LIHUHQoD�QD�GHWHFomRde RANTES entre as culturas
estimuladas e basais dosgrupos de caso e controle.
casos controles-2500
0
2500
5000
7500
10000
P = 0.6595
________________________________________________________Resultados 75
5.5 7HVWHV�GH�FRUUHODomR
��2� tQGLFH� GH� FRUUHODomR� HQWUH� DV� YDULiYHLV� DOHUJLD� H� RV� tQGLFHV� GH�
HVWLPXODomR� HQFRQWUDGRV� FRP� R� WHVWH� GH� SUROLIHUDomR� FRQYHQFLRQDO� IRL� GH�
0,55 (P ����������WDEHOD����QRV�DQH[RV��$�WDEHOD����DSUHVHQWD�RV�tQGLFHV�
GH�FRUUHODomR�HQFRQWUDGRV�HQWUH�DOHUJLD�H�RV�tQGLFHV�GH�HVWLPXODomR�REWLGRV�
com cada citocina testada. A tabela 13, disSRQtYHO�QRV�DQH[RV��PRVWUD�TXH�
QmR�KRXYH�FRUUHODomR�HQWUH�RV�UHVXOWDGRV�GRV�tQGLFHV�GH�HVWLPXODomR�REWLGRV�
FRP� R� 73/� H� FRP� DV� GLYHUVDV� FLWRFLQDV�� 3DUD� WRGDV� DV� DQiOLVHV� GH�
FRUUHODomR�D�YDULiYHO�DOHUJLD�IRL�JUDGXDGD�SHOR�WHVWH�GH�FRQWDWR�HP���������H�
3 que FRUUHVSRQGHUDP�D�SDFLHQWHV��VHP�DOHUJLD��DOpUJLFRV�FRP�XPD�FUX]�QR�
TC, duas cruzes e 3 cruzes, respectivamente.
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VHQVLELOL]DGRV� DR� FURPR� HVWi� FODUR� TXH� HVVH� WHVWH�� QD� VXD� IRUPD�
FRQYHQFLRQDO��QmR�DSUHVHQWD�VHQVLELOLGDGH�VXILFLHQWH�SDUD�VXEVWLWXLU�R�7&�24
���)RUPDV�DOWHUQDWLYDV�GHVVH�PpWRGR, como o uso de CD no lugar dos
PRQyFLWRV�RX�D�LQGXomR�H�D�GHWHFomR�GD�UHVSRVWD�SRU�FLWRFLQDV�FRP�R�LQWXLWR�
GH�PHOKRUDU�D�VHQVLELOLGDGH��UHDEULUDP�DV�SRUWDV�SDUD�QRYDV� LQYHVWLJDo}HV��
Os poucos estudos que seguiram esses protocolos alternativos apresentam
dLIHUHQoDV� FRQVLGHUiYHLV� TXDQWR� D� VXD� PHWRGRORJLD�� GLILFXOWDQGR� D�
FRPSDUDomR� GRV� UHVXOWDGRV�27-32 Mesmo assim, todos apresentaram algum
UHVXOWDGR� HVWLPXODQGR� R� UHILQDPHQWR� GHVVHV� SURWRFRORV� DWUDYpV� GH� QRYDV�
pesquisas.
���'HQWUH�DV�GLIHUHQoDV�HQFRQWUDGDV�QRV�PpWRGRV�DOWHUQDWLYRV� WHPRV��
D� LQGXomR� GD� UHVSRVWD� SRU� PLWyJHQR� RX� FLWRFLQDV�� SHUtRGRV� GLIHUHQWHV� GH�
FXOWXUD��SRXFDV�H�GLIHUHQWHV�FLWRFLQDV�WHVWDGDV�SDUD�GHWHFomR�GR�UHVXOWDGR�H��
HP� XP� WUDEDOKR�� DXVrQFLD� GH� JUXSR� FRQWUROH�� 3DUD� VDQDU� DOJXPDV� GHVVDV�
TXHVW}Hs esse estudo foi realizado comparando a resposta do TPL
FRQYHQFLRQDO� j� GHWHFomR� GH� QRYH� FLWRFLQDV� QDV� FXOWXUDV� GH� �� GLDV� GH� ���
SDFLHQWHV�DOpUJLFRV�DR�FURPR�H����FRQWUROHV�
O IFN- �IRL�HVFROKLGR�SRU�VHU�SURGX]LGR�SHORV�/7F��H�/7K��DQWtJHQR-
HVSHItFLIRV��SULQFLSDLV�FpOXODV�HIHWRUDV�GD�GHUPDWLWH�GH�FRQWDWR�DOpUJLFD.108,111
(VVDV� FpOXODV� Oiberam IFN- � TXH� HVWLPXOD� RXWUDV� FpOXODV� 7�� FpOXODV� 1.� H�
id23720406 pdfMachine by Broadgun Software - a great PDF writer! - a great PDF creator! - http://www.pdfmachine.com http://www.broadgun.com
________________________________________________________'LVFXVVmR 77
PDFUyIDJRV�D�DPSOLDU�R�SURFHVVR�LQIODPDWyULR�H�DXPHQWD�D�H[SUHVVmR�ICAM-
1 e GH�)DV�QR�TXHUDWLQyFLWRV� WRUQDQGR-os maLV�VXVFHWtYHLV�j�FLWRWR[LFLGDGH�
mediada por FasL.80,81,111,140,141
A IL-��p�XP�IDWRU�GH�FUHVFLPHQWR�SDUD�DV�FpOXODV�7�DWLYDGDV��(OD�p��
SURGX]LGD� SHORV� SUySULRV� /7� KDSWHQR-HVSHFtILFRV� DWLYDGRV� H� DWXD� SRU�DomR�
DXWyFULQD� OHYDQGR� D� SUROLIHUDomR� FHOXODU�� SURcesso esse conhecido como
H[SDQVmR�FORQDO�66-68
A IL-���SURGX]LGD�SHOD�FpOXOD�7�1.�LQYDULDQWH��p�QHFHVViULD�SDUD�D�
DWLYDomR� GRV� OLQIyFLWRV� %-�� H� FRQVHT�HQWH� SURGXomR� GD� ,J0� DQWtJHQR-
HVSHFtILFD�� $� DXVrQFLD� GH� ,/-4 diminui a resposta do contato em
camundoQJRV�³knock-out´�SDUD�R�JHQH�GHVVD�LQWHUOHXFLQD�46,160
A IL-�� p� SURGX]LGD� HP� PRGHUDGD� TXDQWLGDGH� SHODV� FpOXODV� 7�
UHJXODGRUDV� DQWtJHQR-HVSHFtILFDV�70 A IL-�� WDPEpP� p� QHFHVViULD� SDUD� D�
PDQXWHQomR�GD�SURGXomR�GD� ,J0��&DPXQGRQJRV�FRP�³knock-out´�GR�JHQH�
da IL-��DSUHVHQWDP�GLPLQXLomR�GD�UHVSRVWD�GH�FRQWDWR�45
A IL-10 foi escolhida por ser produzida em grande quantidade pelas
FpOXODV�7�UHJXODGRUDV�SDUD�VXSULPLU�D�UHVSRVWD�GH�FRQWDWR�70
A IL-1�� p� H[SUHVVD� SHODV� &'� DSyV� D� PDWXUDomR� FRPSOHWD� GDV�
PHVPDV� H� p� QHFHVViULD� SDUD GLIHUHQFLDomR� H�PDWXUDomR� GRV� /7� KDSWHQR-
HVSHFtILFRV�HP�FpOXODV�HIHWRUDV�H�GH�PHPyULD.112
Minang et al.� GHPRQVWUDUDP� TXH� RV� PHWDLV� OHYDP� D� SURGXomR� GH�
FLWRFLQDV�SDGUmR�WDQWR�7K���FRPR�R�,1)- e a IL-2, quanto Th2, como a IL-4 e
IL-13, naV� FXOWXUDV� GRV� SDFLHQWHV� DOpUJLFRV�� $� ,/-13 foi a citocina que
apresentou a melhor resposta em seus ensaios com o cromo, sugerindo
________________________________________________________'LVFXVVmR 78
XPD�PHOKRU�DYDOLDomR�TXDQWR�DR�VHX�SDSHO�QR�GLDJQyVWLFR�GH�DOHUJLD�D�HVVH�
metal.
A IL-17, produzida pelos LTh17 estimula a� SURGXomR� GH� FLWRFLQDV�
SUy-LQIODPDWyULDV��PRGXOD�D�OLEHUDomR�GH�TXLPLRFLQDV�H�DXPHQWD�D�H[SUHVVmR�
de ICAM-�� QRV� TXHUDWLQyFLWRV�� R� TXH� IDYRUHFH� D� LQWHUDomR� FRP� RV�
OLQIyFLWRV�115-117
A quimiocina RANTES�� WDPEpP� FKDPDGD� GH� &&/��� p� SURGX]LGD�
SHOR� HQGRWpOLR�� SODTXHWDV� H� FpOXODV� 7�� (OD� DWUDL� PRQyFLWRV�� &'�� EDVyILORV��
HRVLQyILORV��FpOXODV�1.�H�FpOXODV�7�H�HVWi�HQYROYLGD�QD�DWLYDomR�GRV�/7�66
���&RP� UHODomR� DR� 73/� FRQYHQFLRQDO�� D� UHVSRVWD� DR� ³SRNHZHHG´ foi
adequada e de igual intensidade nos casos e controles; confirmando que as
FRQGLo}HV�GH�FXOWXUD�IRUDP�DGHTXDGDV�H�TXH�FDVRV�H�FRQWUROHV�UHVSRQGHP�
GH�IRUPD�VHPHOKDQWH�D�XP�HVWtPXOR�DRV�TXDLV�DPERV�VmR�VHQVtYHLV��ILJXUDV�
5 a 7).
���&RPR� HVSHUDGR�� DSHQDV� RV� OLQIyFLWRV� GR� JUXSR� GH� SDFLHQWHV�
DOpUJLFRV� IRUDP� HVWLPXODGRV� FRP� R� FURPR� �ILJXUDV� �� H� ���� $� GLIHUHQoD�
HQFRQWUDGD� QD� UHVSRVWD� HQWUH� RV� JUXSRV� OHYRX� D� GLIHUHQoD� QRV� ,(�
encontrados entre os grupos (figura 10).
���&RQVLGHUDQGR� XP� ,(� VXSHULRU� D� �� FRPR� FULWpULR� GH� SRVLWLYLGDGH�� D�
resposta ao teste foi encontrada em apenas 6 dos 18 casos e nenhum
controle, o que corresponde a uma sensibilidade de 33% e especificidade de
100% (tabela 1, nos anexos). Esse valor de corte foi determinado como o
LGHDO� SDUD� GLVWLQJXLU� FDVRV� GH� FRQWUROHV� HP� XP� HVWXGR� SUpYLR� IHLWR� SHORV�
mesmos DXWRUHV�VHJXLQGR�DV�PHVPDV�FRQGLo}HV�GH�FXOWXUD�24
________________________________________________________'LVFXVVmR 79
���$� VHQVLELOLGDGH� DTXpP� GR� GHVHMDGR� p� XPD� FDUDFWHUtVWLFD� SUySULD�
desse teste, quando realizado para detectar alergia ao cromo.24 Todavia, a
VHQVLELOLGDGH�ILFRX�DEDL[R�GR�HQFRQWUDGR�QR�QRVVR�HVWXGR�SUpYLR�(60%) e em
RXWUR�HVWXGR�GH�PHVPD�DXWRULD��FXMRV�GDGRV�QmR�IRUDP�SXEOLFDGRV��HQWUH����
casos e 14 controles onde a sensibilidade foi de 53%.24
���-i�D�HVSHFLILFLGDGH�GR�WHVWH�VH�PRVWURX�H[FHOHQWH��FRPR�QR�QRVVR�
HVWXGR�SUpYLR�������H�VXSHULRU�jTXHOD�HQFRQWUDGD�QR�HVWXGR�QmR�SXEOLFDGR��
que foi de 79%.24� &RPR� RV� FULWpULRV� GH� VHOHomR� IRUDP� RV� PHVPRV�� DV�
culturas foram realizadas seguindo a mesma metodologia e preparadas pela
PHVPD� SHVVRD� HP� WRGRV� RV� FDVRV�� HVVDV� GLIHUHQoDV� GHYHP� VH� GHYHU� DR�
acaso.
���2� tQGLFH� GH� FRUUHODomR� GH� 6SHDUPDQ� PRVWURX� H[LVWLU� FRUUHODomR�
PRGHUDGD�HQWUH�DV�YDULiYHLV�DOHUJLD�H�tQGLFH�GH�HVWLPXODomR�REWLGR�SHOR�73/�
convencional, como visto no estudo de Lindemann et al.32
���2� HVWtPXOR� FRP� FURPR� LQGX]LX� D� SURGXomR� GH� ,1)- , IL-5 e IL-13
apenas no grupo de casos (figuras 12, 13, 27, 28, 42 e 43), determinando
GLIHUHQoD�QRV�,(�H� entre casos e controles (figuras 14, 15, 29, 30, 44 e 45).
���$� VHQVLELOLGDGH� H� HVSHFLILFLGDGH� REWLGDV� FRP� D� GHWHFomR� GH� ,)1-
nesse trabalho foram semelhantes as encontradas nos estudos de Trattner
et al.28 (73,7% e 71,4%) e Moed et al.29 (57% e 86%). Esses pesquisadores
adicionaram fitohemaglutinina e citocinas a fim de melhorar a sensibilidade
GR� WHVWH�� &RPR� RV� UHVXOWDGRV� IRUDP� VHPHOKDQWHV� QRYRV� HVWXGRV� VHUmR�
QHFHVViULRV�SDUD�MXVWLILFDU�R�XVR�GHVVHV�PpWRGRV��DR�PHQRV�SDUD�D�GHWHFomR�
de IFN nas culturas estimuladas com cromo. No entanto, duas ressalvas
________________________________________________________'LVFXVVmR 80
GHYHP�VHU� IHLWDV��R�Q~PHUR�GH�FDVRV�HVWXGDGRV�SRU�0RHG�Ht al.29 foi muito
SHTXHQR�H�R�SHUtRGR�GH�LQFXEDomR�GH�7UDWWQHU�et al.28 foi sensivelmente mais
FXUWR��3DUD�XPD�PHOKRU�FRPSDUDomR�FRP�R�WUDEDOKR�GH�7UDWWQHU�et al.28 um
QRYR� HVWXGR� FRP� XP� SHUtRGR� GH� LQFXEDomR� PDLV� FXUWR� p� QHFHVViULR��
Ackerman et al.30 e Lindemann et al.32� WDPEpP� HQFRQWUDUDP� DXPHQWR� QD�
SURGXomR� GH� ,)1- � QRV� SDFLHQWHV� DOpUJLFRV�� PDV� QmR� FDOFXODUDP� D�
VHQVLELOLGDGH� H� HVSHFLILFLGDGH� GR� PpWRGR�� $SHQDV� 0LQDQJ� et al.31� QmR�
GHWHFWDUDP�GLIHUHQoD� QD� SURGXomR� GHVVD� FLWRFLQD� HQWUH� FDVRV� H� FRQWUROHV��
WDOYH]�SHOD�EDL[D�FDVXtVWLFD�
���$� DFXUiFLD� REWLGD� FRP� D� GHWHFomR� Ge IFN- � QmR� SHUPLWH� D�
VXEVWLWXLomR�URWLQHLUD�GR�7&�SRU�HVVH�WHVWH��0DV��D�GRVDJHP�GH�,)1- pode
VHU�~WLO�FRPR�DOWHUQDWLYD��TXDQGR�R�7&�QmR�SXGHU�VHU� IHLWR�RX�FRPR�H[DPH�
complementar.
���2� WHVWH� GH� FRUUHODomR� GH� 6SHDUPDQ� PRVWURX� H[LVWLU� FRUUHODomR�
moderada entre alergia e a resposta do INF- nas culturas, tabela 12 nos
anexos.
Ackerman et al.,30 demonstraram que as culturas de pacientes
DOpUJLFRV� D� FURPR� SURGX]HP� ,/-5 quando estimuladas com esse metal.
7DPEpP�HQFRQWUDPRV�DXPHQWR�GD�,/-5 e determinamos que esse aumento
p�HVSHFtILFR�SRLV�QmR�IRL�YLVWR�QR�JUXSR�FRQWUROH��ILJXUDV����H������7RGDYLD��D�
VHQVLELOLGDGH� H� HVSHFLILFLGDGH� HQFRQWUDGDV� ILFDUDP� DTXpP� GR� GHVHMDGR�
restringindo o uso isolado da IL-�� FRPR�PpWRGR� GLDJQyVWLFR� GH� DOHUJLD� DR�
cromo. Moed et al.,29�FRQVHJXLUDP�tQGLFHV�GH�VHQVLELOLGDGH�H�HVSHFLILFLGDGH�
PHOKRUHV�� ����� H� ���� UHVSHFWLYDPHQWH�� e� SRVVtYHO� TXH� R� HVWtPXOR� FRP�
________________________________________________________'LVFXVVmR 81
FLWRFLQDV� WHQKD� LQGX]LGR� XPD� PHOKRUD� QRV� UHVXOWDGRV��PDV� D� FDVXtVWLFD� p�
muito baixa podendo esse efeito se dever ao acaso. Assim como com o IFN-
�� KRXYH� FRUUHODomR� PRGHUDGD� HQWUH� DOHUJLD� H� RV� QtYHLV� GH� ,/-5 no
sobrenadante das culturas, r = 0,49 (P = 0,0019).
O desempenho do teste com a IL-13 nesse trabalho foi
surpreendente. Como visto por Minang et al.31�HVWD�FLWRFLQD�HVWi�DXPHQWDda
QRV�SDFLHQWHV�DOpUJLFRV�DR�FURPR�H�SDUD�HVVHV�DXWRUHV�� WDPEpP� IRL�D�TXH�
DSUHVHQWRX� RV� PHOKRUHV� UHVXOWDGRV�� $SHQDV� GRLV� SDFLHQWHV� QmR�
UHVSRQGHUDP� DR� WHVWH� XVDQGR� R� ,(� VHQGR� TXH� XP� GHOHV� WDPEpP� QmR�
UHVSRQGHX�DR� tQGLFH� . Os dois pacientes tiveram resposta de duas cruzes
no TC. No estudo de Minang et al.,31 os 3 pacientes que apresentaram
HULWHPD� H� HGHPD� VHP� D� IRUPDomR� GH� SiSXODV� QR� 7&� - resultado positivo
IUDFR� SDUD� RV� DXWRUHV� H� GXYLGRVR� VHJXQGR� D� QRVVD� LQWHUSUHWDomR� -� QmR�
responderam ao teste. Acreditamos�TXH�D�VHQVLELOLGDGH�GR�WHVWH�ILFRX�DTXpP�
GD� VXD� SRVVLELOLGDGH� SHOD� LQFOXVmR� GH� FDVRV� GXYLGRVRV� Qesse estudo. A
GHWHFomR�GH�,/-���WDPEpP�SDUHFH�SURPLVVRUD�FRPR�PpWRGR�GLDJQyVWLFR�HP�
RXWURV�DQWtJHQRV174,175
���&RQVLGHUDQGR�TXH�DWp�XPD�HP�GXDV�UHDo}HV�SRVLWLvas no TC para o
ELFURPDWR�GH�SRWiVVLR� WHVWDGR�D������VmR�IDOVR-positivas, podemos afirmar
que a especificidade do resultado encontrado com a IL-���p�VXSHULRU�D�GR�
TC.170� $� UHGXomR� GHVVD� FRQFHQWUDomR� SDUD� ������ SRGH� FRUULJLU� HVVH�
problema com o TC, mas leva a uma perda na sensibilidade.12,32,38 Dessa
IRUPD��D�GHWHFomR�GH�,/-13 pode ser considerada adequada para substituir o
________________________________________________________'LVFXVVmR 82
7&�QR�GLDJQyVWLFR�GH�'&$�SRU�FURPR��2� WHVWH�GH�6SHDUPDQ�PRVWURX� IRUWH�
FRUUHODomR�HQWUH�DOHUJLD�H�D�GHWHFomR�GH�,/-13, r = 0,82 e P < 0.0001.
���1mR� KRXYH� GLIHUHQoD� HVWDWtVWLFD� QD� SURGXomR� GH� ,/-2 entre as
culturas basais e estimuladas tanto de casos quanto controles (figuras 16 a
�����$SHVDU�GLVVR�IRL�HQFRQWUDGR�GLIHUHQoD�QRV�,(��PDV�QmR�QR� , de casos e
controles (figuras 19 e 20). Isso SRGH�VHU�H[SOLFDGR�SHOR�DXPHQWR�GD�PpGLD�H�
GD�PHGLDQD�YLVWR� QDV�FXOWXUDV� HVWLPXODGDV��HP� UHODomR�jV�FXOWXUDV�EDVDLV��
GRV�FDVRV�DOLDGR�D�GLPLQXLomR�GD�PpGLD�H�PHGLDQD�GDV�FXOWXUDV�HVWLPXODGDV�
GRV�FRQWUROHV�HP�UHODomR�DV�EDVDLV� �WDEHOD����QR�DQH[R���(VVDV�PXGDQoDV�
QD�SURGXomR�GH�,/-��QmR�IRUDP�GHWHFWDGDV�QD�FRPSDUDomR�HQWUH�DV�FXOWXUDV�
basais com as culturas estimuladas dos dois grupos. No entanto, o discreto
aumento nas culturas estimuladas de casos levou a um aumento equivalente
nos IE desse grupo e a discrHWD� GLPLQXLomR�YLVWD� QDV�FXOWXUDV� HVWLPXODGDV�
GRV� FRQWUROHV� GLPLQXLX� RV� ,(� GRV� FRQWUROHV�JHUDQGR� D� GLIHUHQoD� HVWDWtVWLFD�
encontrada entre os IE dos grupos.
Ackerman et al.30� HQFRQWUDUDP� DXPHQWR� QD� SURGXomR� GH� ,/-2 nas
culturas dos casos quando estimuladRV�FRP�ELFURPDWR�GH�SRWiVVLR��0LQDQJ�
et al.31� WDPEpP� HQFRQWUDUDP� DXPHQWR� GD� ,/-�� FRP�(/,6SRW�PDV� QmR� FRP�
(/,6$��-i�/LQGHPDQQ�et al.,32�XWLOL]DQGR�(/,6SRW��QmR�REVHUYDUDP�DXPHQWR�
QD� SURGXomR�GH� ,/-���$SHVDU� GH� QmR� GHWHFWDGR�QHVVH� HVWXGR� p� SRVVtYHO� H�
OyJLFR�TXH�RFRUUD�XP�DXPHQWR�QD�SURGXomR�GH�,/-2, mas esse aumento, se
H[LVWLU�� p� LQVXILFLHQWH� SDUD� VHSDUD� FDVRV� GH� FRQWUROHV�� SHOR� PHQRV� QDV�
culturas de 6 dias.
________________________________________________________'LVFXVVmR 83
���$�DFXUiFLD�GR�WHVWH�FRP�R�,(�QmR�HQFRUDMD�R�VHX�XVR�QR�OXJDU�GR�7&�
SDUD�R�GLDJQyVWLFR�GH�DOHUJLD�Do cromo, especialmente devido a sua baixa
sensibilidade (61%). No estudo de Minang et al.31�D�VHQVLELOLGDGH�WDPEpP�IRL�
um problema, apenas 2 dos 9 pacientes - seis se excluirmos os 3 pacientes
com apenas eritema e edema no TC - responderam com aumento da IL-2.
Como QmR�KRXYH�GLIHUHQoD�HQWUH�RV� de casos e controles a sensibilidade e
D� HVSHFLILFLGDGH� SDUD� HVVH� tQGLFH� QmR� IRUDP� FDOFXODGas. Foi encontrado
FRUUHODomR� PRGHUDGD� HQWUH� DOHUJLD� H� RV� QtYHLV� GH� ,/-2 produzidos nas
culturas, r = 0,34 e P = 0,0410.
2V�FDVRV�H�FRQWUROHV�DSUHVHQWDUDP�GLIHUHQoD�QD�SURGXomR�EDVDO�GH�
IL-��� R� TXH� LPSHGH� TXDOTXHU� FRPSDUDomR� HQWUH� RV� JUXSRV� �ILJXUD� ����� 2�
PRWLYR� GHVVD� GLIHUHQoD� QmR� HVWi� FODUR�� 1R� HVWXGR� GH� $FNHUPDQ� et al.30 o
FURPR� LQGX]LX� D� SURGXomR� GH� ,/-4 nos pacientes alpUJLFRV�� R� TXH� QmR� IRL�
encontrado nesse estudo. Percebemos aumento da IL-4 no grupo controle,
PDV�QmR�QRV�FDVRV��ILJXUDV����H������/LQGHPDQQ�et al.32 e Minang et al.31
QmR�HQFRQWUDUDP�GLIHUHQoD�QD�SURGXomR�GH�,/-��HQWUH�SDFLHQWHV�DOpUJLFRV�H�
QmR�DOpUJLFRV�
���6HJXQGR� QRVVRV� DFKDGRV�� SRGHUtDPRV� LPDJLQDU� TXH� D� ,/-4
DSUHVHQWD�XP�HIHLWR�VXSUHVVRU�QD�'&$�SHOR�FURPR��HIHLWR�HVVH�TXH�QmR�HVWi�
SUHVHQWH�QRV�LQGLYtGXRV�DOpUJLFRV�RX�TXH�HOD�VHMD�SURGX]LGD�FRPR�UHVSRVWD�D�
RXWUR� IDWRU� JHUDGR� SHOD� SUHVHQoD� GR� FURPR� QD� FXltura. Os outros estudos
DSUHVHQWDUDP� UHVXOWDGRV� GLIHUHQWHV�� PDV� R� SHUtRGR� GH� FXOWXUD� GHVVHV�
WUDEDOKRV� IRUDP� PDLV� FXUWRV�� YDULDQGR� GH� ��� D� ��� KRUDV�� D� H[FHomR� GR�
estudo de Ackerman et al.,30 QR�TXDO� WDO� LQIRUPDomR�QmR�HVWD�GLVSRQtYHO��$�
________________________________________________________'LVFXVVmR 84
GLQkPLFD� GDV� FLWRFLQDV� QHVVDV� FXOWXUDV� GLILFXOWD� D� FRPSDUDomR� HQWUH� RV�
resultados.175
���2�HVWtPXOR�FRP�FURPR�GHWHUPLQRX�XPD�UHGXomR�QD�SURGXomR�GH�,/-
��� QRV� GRLV� JUXSRV� �ILJXUD� ���H������ $� UHGXomR�GH� ,/-10 ocorreu nos dois
JUXSRV� ID]HQGR� FRP� TXH� QmR� H[LVWD� GLIHUHQoD� QRV� FULWpULRV� GLDJQyVWLFRV�
entre eles (figuras 34 e 35). Lindemann et al.32� QmR�HQFRQWUDUDP�GLIHUHQoD�
QD�SURGXomR�GH�,/-���HQWUH�SDFLHQWHV�DOpUJLFRV�H�QmR�DOpUJLFRV��Mi�$FNHUPDQ�
et al.,30� HQFRQWUDUDP� DXPHQWR�� 3RVVLYHOPHQWH� HVVDV� GLIHUHQoDV� VH� GHYHP�
DRV�SHUtRGRV�GH�FXOWXUD�HPSUHJDGRV�RX�D�GLIHUHQoD�QDV�FRQFHQWUDo}HV�H�QD�
IRUPD�GR�FURPR�XWLOL]DGD��1HVVH�HVWXGR�D�FRQFHQWUDomR�GH�FURPR�XVDGD�IRL�
superior aos dos demais estudos o que pode ter gerado dano celular e esse
VLQDO�GH�SHULJR�VXSULPLGR�D�DomR�GDV�FpOXODV�UHJXOadoras, diminuindo assim
D�OLEHUDomR�GH�,/-10.
���1mR� KRXYH� GLIHUHQoD� HVWDWtVWLFD� QD� SURGXomR� GH� ,/-12 entre as
culturas basais e estimuladas tanto de casos quanto controles (figuras 36 a
38). Lindemann et al.32� WDPEpP�QmR�HQFRQWUDUDP�GLIHUHQoD�QD�SURGXomR de
IL-���HQWUH�SDFLHQWHV�DOpUJLFRV�H�QmR�DOpUJLFRV��7RGDYLD��$FNHUPDQ�et al.,30
GHWHFWDUDP�DXPHQWR�QD�SURGXomR�GH�,/-12 em suas culturas.
$SHVDU�GH�QmR�WHUPRV�REVHUYDGR�DXPHQWR�QD�SURGXomR�GH�,/-12 nos
FDVRV� IRL� HQFRQWUDGR� GLIHUHQoD� QRV� tQGLFHV� GH� HVWLPXODomR� H� QR� GHOWD� GH�
casos e controles (figuras 39 e 40). De forma semelhante ao discutido com a
IL-��� LVVR� VH� GHYH� DR� DXPHQWR� GD� PpGLD� H� GD� PHGLDQD� HQFRQWUDGDV� QDV�
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GLPLQXLomR�GD�PpGLD�H mediana das culturas estimuladas dos controles em
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do teste encontrada tanto com o IE quanto com o �QmR�Ioi adequada para
que ele substitua o TC, mas pode ser usado como alternativa.
��1mR�KRXYH�GLIHUHQoD�QD�SURGXomR�GH�,/-17 entre as culturas basais
e estimuladas de casos e controles (figuras 46 a 48) nem nos valores
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produzir RANTES quando cultivados com cromo (figuras 52 e 53). A
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GLDJQyVWLFRV�GRV�JUXSRV�
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A IL-13 apresenta adequada sensibilidade e especificidade para o
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���$�GHWHFomR�GH�,)1- , IL-5 e IL-12 tanto pelo IE quanto pelo e de
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desses�H[DPHV�QmR�p�VXSHULRU�D�GR�73/�FRQYHQFLRQDO�
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capacidade desse teste.
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ANEXOS
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___________________________________________________________Anexos 87
8 ANEXOS
Quadro 2 ±�*UXSR�GH�VXEVWkQFLDV�WHVWDGDV�QD�EDWHULD�SDGUmR� brasileira de dermatite de contato.1 Antraquinona 16 Neomicina2 %iOVDPR-do-peru 17 Nitrofurazona 3 %HQ]RFDtQD 18 Parabeno-mix4 %LFURPDWR�GH�SRWiVVLR 19 Parafenilenodiamina5 Butilfenol-SDUDWHUFLiULR 20 Perfume-mix6 Carba-mix 21 PPD-mix 7 Cloreto de cobalto 22 Prometazina 8 &RORI{QLR 23 Propilenoglicol9 Etilenodiamina 24 Quartenium 15
10 )RUPDOGHtGR 25 Quinolina-mix11 Hidroquinona 26 Resina-HSy[L 12 Irgasan 27 6XOIDWR�GH�QtTXHO 13 Kathon CG 28 Terebintina 14 Lanolina 29 Timerosal15 Mercapto-mix 30 Tiuram-mixFONTE: Grupo brasileiro de estudo em dermatite de contato (GBEDC). (VWXGR�PXOWLFrQWULFR�SDUD�D�HODERUDomR�GH�XPD�EDWHULD-SDGUmR�EUDVLOHLUD�GH�teste de contato. An Bras Dermatol. 2000; 75:147-56.9
Quadro 3 ±�*UXSR�GH�VXEVWkQFLDV�WHVWDGDV�QD�EDWHULD�GH�FRVPpWLFRV�
1 *HUPDOO������,PLGD]ROLGLQLOXUpLD�
2 BHT (Butil hidroxi-tolueno)
3 5HVLQD�WRQVLODPLGD�IRUPDOGHtGR
4 Trietanolamina
5 Bronopol (Bromo-2-nitropropano-1,3-diol 2)
6 Cloracetamida
7 ÈFLGR�6yUELFR
8 7LRJOLFRODWR�GH�DP{QLR
9 Amerchol L-101
10 Clorhexidine
FONTE: %DWHULD� SDGUmR� GD IPI-ASAC Brasil , Rio de Janeiro, Brasil. Em: http://www.ipibrasil.com.br/v2/pages/parcerias_endoderme_contatoCosmeticos.asp?catProdAtual=1.
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___________________________________________________________Anexos 88
Quadro 4 - Resultados dos testes de contato Casos Teste de contato Controles Teste de contato 1 4 (++), 6, 27 e 30 1 Negativo 2 4 (++) e 6 2 Negativo 3 4 (++), 6 e 30 3 Negativo 4 4 (++), 6 e 7 4 Negativo 5 4 (++) e 6 5 8, 20 e 306 4 (++) e 25 6 27 e 10c7 4 (++), 6 e 30 7 20, 27 e 8c8 4 (++) e 27 8 27 e esmalte 9 4 (++) 9 9 e 1610 4 (+++), 7, 17 e 30 10 8 e 1011 4 (+) e 6 11 Negativo 12 4 (+++) e 27 12 2713 4 (++) 13 17 e 2914 4 (+++), 7, 27 e 30 14 3 15 4 (++) 15 1616 4 (+++), 10 e 27 16 7, 22 e 2717 4 (++) e 6 17 Negativo 18 4 (++) e 21 18 Negativo 19 7 FONTE: Dermatologia do HC-FMUSP NOTA:� 2V� Q~PHURV� DSUHVHQWDGRV� QDV� FROXQDV� GRV� WHVWHV� GH� FRQWDWR�correspondem aos utilizados pelas baterias de teste de contato SDGUmR�H�GH�FRVPpWLFRV� H� HVWmR� GLVSRQtYHLV� QR� DQH[R�� TXDGURs 2 e 3. Quando os Q~PHURV� IRUHP� VHJXLGRV� GD� OHWUD� ³F´� LQGLFD� TXH� SHUWHQFHP� D� EDWHULD� GH�FRVPpWLFRV�$�LQWHQVLGDGH�GD�UHDomR�DR�ELFURPDWR�GH�SRWiVVLR�QDV����KRUDV�IRL�GHVFUita como(+) ± positivo fraco; (++) ± positivo forte; (+++) ± positivo muito forte (+++).
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____________________________________________5HIHUrQFLDV�ELEOLRJUiILFDV100
9 5()(5Ç1&,$6�%,%/,2*5È),&$6
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