FUNCTIONAL INVESTIGATION OF FUNCTIONAL INVESTIGATION OF ANTIBODY PRODUCTION IN COMMON ANTIBODY PRODUCTION IN COMMON VARIABLE IMMUNODEFICIENCY (CVID) VARIABLE IMMUNODEFICIENCY (CVID) PATIENTS UNDER IMMUNOGLOBULIN PATIENTS UNDER IMMUNOGLOBULIN

SUBSTITUTIONSUBSTITUTION

Zita TrávníčkováZita TrávníčkováDepartment of Clinical Immunology and Allergology of St. Anne´s

University Hospital and Medical Faculty of Masaryk University Brno

ESID Prague Spring Meeting 2009

ELISPOT assay in PIDELISPOT assay in PIDfunctional investigation

of B cells

Detection of specific antibody production on B cell level

independent on substitution Ig therapy

EXPERIMENTAL DESIGNSUBJECTSSUBJECTS

37 CVID patients (15 males, 24 females, age range 20 - 74 years)

81 healthy donors (28 males, 53 females, age range 14 - 74 years)

VACCINATIONVACCINATION ALTEANA (Sevapharma a.s., Prague, Czech Republic) PNEUMO 23 (Sanofi Pasteur, Lyon, France)

ELISPOTELISPOT (isolated MNC of peripheral blood, calculated on CD19+)

day 0, day 7, weeks 4-11

HEALTHY DONORS

Vaccination with Vaccination with PROTEIN antigenPROTEIN antigen

50 controls (16 males, 34 females, age range 22 – 72 years)

Vaccination with Vaccination with POLYSACHARIDE antigenPOLYSACHARIDE antigen

10 controls ( 4 males, 6 females, age range 15 – 46 years)

Vaccination with Vaccination with PROTEIN and POLYSACHARIDE antigenPROTEIN and POLYSACHARIDE antigen

21 controls (8 males, 13 females, age range 14 – 50 years)

CVID PATIENTS

Vaccination with Vaccination with PROTEIN and POLYSACHARIDE antigenPROTEIN and POLYSACHARIDE antigen

37 patients (15 males, 24 females, age range 20 - 74 years)

EUROclassB-cells

1% B cells group B-

1% B cells group B-

> 1% B cells group B+

> 1% B cells group B+

2% switched memory B cells group smB-

2% switched memory B cells group smB-

2% switched memory B cells group smB+

2% switched memory B cells group smB+

< 10% CD 21low B cellsgroup smB-21norm

< 10% CD 21low B cellsgroup smB-21norm

10% CD 21low B cellsgroup smB-21lo

10% CD 21low B cellsgroup smB-21lo

10% CD 21low B cellsgroup smB+21lo

10% CD 21low B cellsgroup smB+21lo

< 10% CD 21low B cellsgroup smB+21norm

< 10% CD 21low B cellsgroup smB+21norm

(Blood 2008)

EUROclassB cells

group B-group B-group B+37 patients37 patients

group B+37 patients37 patients

group smB-

21 patients21 patients group smB-

21 patients21 patients group smB+

16 patients16 patients

group smB+

16 patients16 patients

group smB-21norm

9 patients9 patientsgroup smB-21norm

9 patients9 patients

group smB-21lo

12 patients12 patientsgroup smB-21lo

12 patients12 patientsgroup smB+21lo

10 patients10 patientsgroup smB+21lo

10 patients10 patients

group smB+21norm

6 patients6 patientsgroup smB+21norm

6 patients6 patients

CVID

CVIDBEFORE VACCINATION

CVIDDAY 7 AFTER VACCINATION

CVID4-11 WEEKS AFTER VACCINATION

CVIDBEFORE VACCINATION

CVIDDAY 7 AFTER VACCINATION

CVID4-11 WEEKS AFTER VACCINATION

CONCLUSIONSCONCLUSIONS

The ELISPOT assay is sensitive functional test for sensitive functional test for determination of specific antibody production in PIDPID

patients under substitution immunoglobulin therapyunder substitution immunoglobulin therapy.

In well-defined CVID patients almost no detectable almost no detectable peripheral B-cells producing specific IgG, IgA and IgM peripheral B-cells producing specific IgG, IgA and IgM antibodiesantibodies agains Tet. Tox. and PCP were observed.

IfIf the IgG response was detectable was detectable after Tet. Tox. vaccination most patients but not all were characterized

as smB+21smB+21norm norm according to EUROclassaccording to EUROclass.

SPECIAL THANKS TO CO-WORKERS

Department of Clinical Immunology and AllergologySt. Anne´s University hospital and Medical Faculty of

Masaryk University in Brno

prof. MUDr. Jiří Litzman, CSc.prof. MUDr. Jiří Litzman, CSc.MUDr. Vojtěch Thon, Ph.D.MUDr. Vojtěch Thon, Ph.D.Mgr. Marcela Vlková, Ph.D.Mgr. Marcela Vlková, Ph.D.

1910

2009

St. Anne´s University Hospital Brno