Human Basophil Releasability. II. Changes Basophil Releasability in Patients with Atopic Dermatitis

. In

Gianni Marone, M.D., Roberto Giugliano, M.D., Giuseppe Lembo, M.D., and Fabio Ayala, M.D. Departments of Medicine and Dermatology , University of N aples, II School of Medicine, N aples, Italy

" R eleasability" is the theory w hereby bioche mica l events in basophils influence the capacity to release che mical m e­diators in response to activating stimuli . W e have compared the releasability o f basophils fro m 21 young patients w ith atopic dermatiti s (AD) with that fro m 17 normal dono rs of m atched ages. B asophils were chaUen ged with several different stimuli : rabbit antihuman Fc. (anti-IgE), N-fo r­myl-methionyl-Ieucyl-phenylalanine (f-met-peptide), C a + + ionophore A23187, and D 20 . Basophils from patients with AD released sig nificantly m o re histamine bo th "s po nta­n eously" and in response to D 20 than did controls. The basophils of p atients with AD w ere sig nifi cantl y m o re re-

A topic dermatitis (AD) is a common fa milial, pruritic skin diso rder that begins during ea rl y in fa ncy and is characterized by several immunologic abnormalities [1] . Although the pathogenic impo rtance of IgE in the inflammatory response of AD remains un certain ,

serum concentrations of IgE are elevated in =80% of patients with AD [2,3]. A common ass umption holds that IgE plays a rol e in AD by diffusing into skin , binding to baso phils and/o r mast cells, and then, when the IgE is bridged by a specifi c anti gen, triggerin g the release of histamine and other chemica l mediators to initiate the inflammatory response [1]. Although the specifi c antigens underlyin g the pathogenesis o f AD have yet to be iden­tified, an increased number of mast cells [4,5] and elevated his­tamine concentration in the skin have been demonstrated in AD [6, 7] . Recent studies have attributed a pathogenic role to an IgE­mediated response of basophils in patients w ith AD [8-1 0].

In vitro histamine release from basophils is increased [11- 13] and , under appropriate circumstances , plasma histamine is en­hanced in patients with AD [14] . These findin gs suggest that the pathogenesis of AD might involve abnormal mediator release (increased "releasability"), poss ibly in response to endogenous/ exogenous stimuli .

Manuscript received July 16, 1985; accepted fo r publica tion January 2, 1986.

Supported by g rants from the C. N .R. (83.00430.04,84.01756.04, and 85.00491.04) and the M .P.1. (Ro me, Italy) .

Reprint requests to: Gianni Marone, M .D. , I C linica Medica, II Facolta M edicina, Via S. Pansini 5, 80131 N apoli , Italy.

Abbreviations: AD: atopic dermatitis anti-IgE: rabbit anti-Fc. Con-A: concanavalin A DzO: heavy water f-m et-peptide: N-form yl-methionyl- Icucyl-phenylalaninc P : PIPES buffer 25 mM , N aCI 11 0 111M, KCl 5 111M , pH 7.35 PC: PIPES bu ffer containing 1.0 111M CaCh PIPES: piperazine-N, N' -bis(2-ethanesulfonic acid)

spo nsive to anti-fgE and to A23187. T here w as no di ffer­en ce between the percent f-met-peptide-induced histamine release in patients with AD vs contro ls. No signifi cant correlati o n between percent histamine release with optimal o r subo ptimal concentrati o ns of the stimuli and serum IgE level was fo und . There was a sig nifi cant co rrel ation be­tween the sensiti v ity o f the cell s to release with f-met­peptide and the respo nse to A23187 both in contro l and in AD patients . Since basophils are tho ught to play some role at the site of inflammatio n in AD, their increased releas­ability might contribute to the sy mptoms of these patients. ) In vest Dermatol 87: 19-23, 1986

Evidence is accumulating in favo r of a releasability para meter. Basophil releasability implies that biochemi cal events, not only the surface density of IgE molecules, determine the capacity of basophils to release mediators in response to activating stimuli . Basophil releasability is signifi ca ntl y correlated with cell donor age [15 ,16] and is altered in patients with chronic urtica ria [17 ,18] and with as thma [1 9,20].

The present experiments were conducted to ascertain whether there were any differences in the releasability of basophils from youn g patients with AD compared with those from controls of matched ages. To probe the release mechanism in detail , we used a variety of rel easing agents : an IgE-specific stimulus, rabbit antihuman IgE (anti-IgE); N-form yl-methionyl-leucyl-phenyl­alanine (f-m et-peptide), which interacts with a specific cell surface receptor independent o f the IgE receptor [21 ]; the Ca + + iono­phore A23187, w hich bypasses some of the ea rl y stages of the release process and causes release by allowing transmembrane calcium influx [22]; and an ampli fy ing agent, heavy w ater (D20) , which in vitro potentiates the release of mediators induced by other stimuli [21], bu t ca uses little release by itself in normal dono rs [20].

The results demonstra te that the maximum percent histamine release and cell sensitivity to a suboptimal concentration of both anti-lgE and A23187 arc significantly increased in young patients with AD . The ability to release histamine "spontaneously" and in res ponse to D20 was also increased. These abnormalities w ere independent of serum IgE and may re fl ect an impaired regulation of releasability .

MATERIALS AND METH ODS

Reagents Piperazine-N , N' -bis(2-ethanesulfonic acid) (PIPES) was purchased fro m Sigma C hemical Co., St. Louis, Missouri. Rabbit antihuman IgE (anti-l gE) directed aga inst the Fc. portion of E myeloma protein was generously provided by Dr. K. Ish­izaka, The Johns Hopkins University, Baltimore, Maryland . The fo rmylated tripeptide (f-met-peptide) and the Ca + + ionophore

0022-202X/86/$03.50 Copyright © 1986 by T he Society for In ves tigative Dermatology , Inc.

19

20 MAnONE ET AL

A23187 were obtai ned from C albiochem-Behring Corp. , LaJolla, Californ ia.

Buffers T he PIPES buffers used in thl:se experiments were made up of 25 mM PIPES, pH 7.35 , 110 mM N aC I, 5 mM KCI. This mix ture is referred to as P; PC con tain s, in add iti on to P, 1. 0 mM CaCI2 [23,24) .

Subjects Twent y-one young patients, ages 6 months to 18 years (m ea n , 5.6 ± 0.9 years), w ith documentated m oderate to severe AD were studied . Seventeen normal individuals, ages 5-18 years (mean 7 ± 0.6 years) wi th no personal histo ry of any form of ato pi c disease served as cont ro l subjec ts. Donors received no sys­temic or topical medi cation for 1 week prior to the study.

Cell Suspension and Histamine Release After in for med con­sent was obtained, approx imately 25 ml of blood were drawn; 2 ml of blood were saved for serum IgE determinations , and the rest was drawn into a mi xture of 0 .008 M EDT A, 1.1 % Dextran 70, and g lucose . The erythrocytes were allowed to settle for 90 min at 22°C, and leukocyte- rich plasma was removed. The cell s we re separated by centri fugation at 180 g for 8 min , washed twi ce in P, and resuspended in Pc. Basophils w ere co unted in a Spiers­Levy chamber after staining w ith Alcian blue accord ing to the method of G ilbert and O rn stein [25]. Aliquots (0.4 ml) of the cell suspension w ere placed in Falcon 12 X 75 mm polyethylene tubes and warmed to 37°C; 0.2 ml of each prewarmed stimulus was added, and incubation was co ntinued at 37°C for 45 min . After cen tri fugat io n , the cell-free supernatant was assayed for histam ine w ith an auto m ated Au orometric technique [26,27]. The net per­centage histam ine released by secretagogues was calcul ated usin g the total hi stamine release from cell aliquo ts lysed with 2% per­chloric acid corrected for the histamine released spontaneously fro m the unstimulated aliquots. The difference between replicate hista mine measurements was less th an 10%. The anti-IgE, f-met­peptide, and A23187 used throughout th e stud y were aliquots of a single large preparation that had been sto red at - 20°C. Leu­kocytes from a single individu al showed little var iat ion «10%) in histam ine release when cha ll enged on several occas io ns w ith the sa me concentra tion of D 20, anti- IgE, f-met-peptide, and A23187 fro m January through to March 1984.

IgE Measurement Serum samples were stored at - 70°C from the time of collectio n until anal ysis. T he serum IgE levels were determined using a previously described radioimmunoassay [28]. All serum samples were measured in duplicate wi thin the sa me assay.

Statistical Analysis The results we re expressed as the mean ± SEM. The data were analyzed by Student's I-tes t. T he Rank correlation was calculated by the Spearman Rank Coefficient (Y,) [291·

RESULTS

BasophiJs from 17 norm al donors and 21 young patients w ith AD of matched ages (5.6 ± 0.9 vs 7 ± 0.6 years; p > 0.05) were co mpared w ith respect to the tota l histamine content per basophil and the m ean serum IgE level (Ta ble I). The m ean serum IgE level was 709 ± 352 in AD pati ents and 179 ± 48 ng/ml in

Table I. Serum IgE Level and Basophil Hi stamine Content

Controls

Atopic dermatitis

Values arc means ± SEM.

Serum IgE (ng/ml)

179 ± 48 (90-789)

709 ::!: 352 (17-7600)

Numbers in parenthesis represent the range.

Hista mine (pg/basophil)

0.75 ::!: 0.16 (0.27-1. 46)

0.68 ::!: 0.09 (0.30-1. 52)

60

w 50 Vl « w -' ~ 40

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O

THE JOUnNAL OF IN VESTIGATIVE DEnMATOLOGY

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CONTROLS ATOPIC DERMATITIS

I I I

F"=~~.'=l

CONTROLS

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ATOPIC DERMATITIS

Figure 1. A, "Spontaneous" histamine rel ease in 17 normal donors (op e'l circle) and 21 patients with atopic dermatitis (closed circle). Leukocytes were incubated in regular (H20) buffer for 45 min at 37°C. p < 0. 05 . B, 0 20-induced hista mine release in 17 normal donors (OpCII circle) and 21 patients with atopic dermatitis (closed circle). Leukocytes were incubarcd in 44 % 0 20 for 45 min at 3rc. The percent histamine secretion in 0 20 was calculated by subtracting the percent release in regular (H 20) buffer. p < 0.05. Each symbol represents the mean of duplicate determination s. The shaded area represents the mean ::!: SEM.

con tro l su bjects (p > 0.05). The total histamine content in AD patients and contro l subj ects was no t signifi can tly different.

We first exa mined the "spontaneous" release of hi stamine dur­ing 45-min culture in stand ard buffer from basophils of AD and control subj ects. The percent histamine release in AD was 14. 3 ± 2.3vs7. 1 ± 0.7in controls(p < 0 .05) . The "spontaneous" releasability in AD and in controls is illustrated in Fig 1 A. These data are su mmarized in Table II. N o sign ifi ca nt correlat ion be­tween serum IgE level and the percent spon taneous hi stamine release was found in AD (r, = 0.07; P > 0.05) or in con trols (Y, = 0.40; P > 0.05). We next examined the releasabi lity ofbasophils suspended in buffer contain in g 44% D 20. Fig 1B and Table II show that AD patients are signifi can tly more responsive to D20 than are con tro ls (14.6 ± 3.8 vs 3.4 ± 0.6; p < 0.05). T here was no signifi cant correlation between serum IgE level and th e percent histamine release with D 20 either in AD (Y, = 0.00; p > 0.05) o r in controls (r, = 0 .1 6; P > 0.05).

Basophils ob tained frolll no rm al donors and AD patients were also exposed to a ran ge of concentrations ( 1 0 ~ 2 to 1 J-Lg/ml) of ant i-I gE previously shown to be optimal for hista mine re lease [22, 23). T herefo re, an entire dose-response curve was o btained for each individual donor. Table II shows that the m aximum percent anti-I gE-induced histamin e secretion in AD ,vas 35.6 ± 3.8 vs 6.6 ± 1.9 in controls (p < 0.001). The increased IgE­m ediatcd releasability in AD is illustrated in Fig 2. The percent hi stamine release was signifi cantl y hig her in the AD gro up than in the control group, at all con centrati ons of anti-IgE. Aga in, there was no correlation between serum IgE level and the percent histaminc release w ith anti-I gE, either in AD (Y, = 0.02; P > 0.05) or in controls (r, = -0.11 ; p > 0.05).

To stud y releasabilit y in response to the f-met-peptide, which ac tivates a specifi c cell surface recep tor independent of the IgE receptor [21], basophil s were challenged with a range of concen­trations of f-met-peptide (10 - H to 10 - 5 M) previously shown to be o ptim al for histaminc release [1 6,21] . Although there was an increase in the percent histamine release at all f-m et-peptide con­centrations in AD, the difference between patients an~ controls was not signifi cantly different. The m aximum percent f-met­peptide-induced histamine secretion in AD (36.6 ± 4.8) was not statistica lly different from that o btained in control s (27.9 ± 4.2) . There was also no correlation between serum IgE level and the m aximum percent histamine release with f-mct-peptide in con-

VOL. 87. N O. I JULY 1986 B ASOPHIL RELEASABILIT Y IN ATO PIC DERM AT IT IS 21

Table II. M ax imum Pcrcent Histamine Release in Contro ls and Atopic D ermatitis Patients

Controls

"Spontaneous" Histamine Release

Ato pic dermatitis 7. 1 :±: 0.7

14.3 :±: 2.3" 3.4 ± 0.6

14.6 :±: 3.8"

"I' < 0.05 when comp",cd with controls. hI' < 0.001 whcn compared with contro ls.

troIs (r, = - 0.41; P > 0 .05) and in A D patients (r, = 0. 13; P > 0 .05) (d ata no t shown).

With th e Ca + + io no pho re A23187 used at a range o f concen­trati ons (3 X 10 - 2 to 1 J.Lg/ml) prcvio usly shown to be o ptim al fo r ac tivati on o f human baso phil s [1 6,30], the maximum percent of A23187-in duccd hi stamine secreti on in AD (91.4 :±: 2.7) was signifi ca ntl y (p < 0.05) hi gher th an in con trols (80.0 ± 3.8) . Fi g 3 shows th at there is a signifi ca nt increase in the percent histamin e release at all concentrations o f A23 187 in the g ro up of AD vs controls. Again , th ere w as no correla tion betwce n th e se rum Ig E level and the m aximum perccnt A23187-induced hista mine release either in controls (r, = 0.08; p > 0.05) o r in A D pat ients (r, = 0. 12; P > 0.05).

Finall y, th e maximum perccnt histamine release w ith o ptim al concentrati ons o f th e 4 agents (a nti-l gE, D 20, f-m et- peptid , and A23187) was compared. There w as no correlati o n between the sensiti vity o f th e cell s o f the patients with AD to release with anti-fg E and their response to D 20 (r, = 0 .14; P > 0.05) , f-met­peptide (r, = 0.24; P > 0.05), o r A23187 (r, = 0 .1 7; P > 0.05). Sim il arl y, th ere was no co rrelati on between the maximum percent hi stamin e rel ease by f-m et-peptide and D 20 (r, = 0.07; P > 0.05) . Similar results were o btained in the g ro up o f control subj ects, in ag reem ent w ith previous repo rts [1 6, 19]. In contrast, we found a signifi cant co rrelatio n between th e sensitivity o f th e cells to release w ith f-m ct-peptide and th eir rcs ponse to A23187 bo th in con trols (r, = 0.52; p < 0.05) and in AD pa tients (r, = 0.49; p < 0.05) (Fig 4).

DISC U SS lON

Bo th "spontaneous" rel easa bility and basophil sensitivity to im­mun ologic (anti-l gE) and nonimmunologic (D 20 and A23187) stimuli are sig nifica ntl y increased in yo un g patients w ith AD compared w ith age-m atched contro ls. The m aximum percent histamin e release induced by all the stimuli used is unrel ated to the se rum IgE level of the do no r. The maximum histamine release

40 w If)

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./ I z ~ 20 ;:! ~ ! :r: ./ I- 10 z w u ! --...£-----2 a::: _-2--w Cl..

0 n_ .n- ..... .",... 10-2 10-1

ANTI- IgE (}lg/mll

Figure 2. Histamine release as a fun ction of anti- lgE concentration in 17 normal donors (opell circle) and 21 patients with atopic dermatitis (closed circle) . Values are expressed as mean :±: SEM. **Significanti y different from the corresponding concentration of anti- lgE of the group of control subjects (p < 0.001) .

Al1ti-l gE

6.6 ± 1. 9 35.6 :±: 3.8h

(-met-peptide

27.9 :±: 4.2 36.6 ± 4. 8

A23187

80.0 :!:: 3.8 91.4 :!:: 2.7"

induced by the f-m et-peptide is no t signifi cantl y hi gher in AD pa tients th an in co ntro ls. There is a pos itive co rrelation b etween hi stamin e release ca used by the f-m et-peptide and A 23187 in AD and co ntro ls.

The first o bserva tion to emerge fro m this study is th a t w ashed baso phils fro m AD patients release m ore histamine du ring 45-min cul ture in standard bu ffe r than do th ose fro m con t rols. A spon ta neo us h y perreleasable state was clea rl y foun d in approxi­m J tely 50% of AD patients and it was unrelated to serum IgE level s o r to th e release in res ponse to othcr stimuli . A simil ar response has been repo rted in patients with food all e rgy [31], however this findin g has not been confirmcd by o thers [32,33]. Altho ugh AD pa tients w hen challenged w ith appro priate food releasc histamine [1 4J, no co nclusions ca n yet be drawn o n the nature of th e h y perreleasa ble sta te found in so me AD indi v iduals.

A second findin g is tlut , co mpared w ith contro ls , youn g pa­tients w ith AD had a signifi ca ntl y increascd res po nse to D 20. The D20 respo nse did no t correlate w ith th e respo nse to o ther stimuli o r w ith serum IgE levels. D20 cnhances basophil hista­min e release induced by anti gcn and anti- lgE [21,34] and C5a [35], bu t no t f-m et-peptide-ind uced basophil histamine release [21] . D 20 induces thc releasc 'J f hista mine from baso phils of all erg ic do no rs [1 9,20] and it has been sugges ted th at th e " D20 res ponder" basophils represent an enh anced sta te o f baso phil ac­ti va ti on ca used by in vivo expos ure of basop hils to subo ptimal concentrati ons of releas ing agent (s) [36]. Altho ugh the hi s tamine Content per baso phil was similar in AD patients and control sub­jects, it may be speculated that baso phils fro Ill so me pa tients w ith AD are in vivo "activated " by unknown stimuli. This hy p othes is is, at leas t in part , supported by the recent observa ti on that serum concentratio ns o f histamine are eleva ted in patients w ith AD after food challenge [1 4]. We have recentl y fO llnd increased intes tin al permea bility to lactulose in patients w ith AD [37], w hich su ggests tha t alle rgens m ay be rapidl y absorbed , and m ay trigger m ediato r rel ease fro m basophils and skin mast cell s.

100

UJ Ul <l: ~ 80 UJ cr

~ 60 ~ <l: I-

~ 40

l-z UJ ~ 20 w CL

0

* ..

10-2 10-1

A23187 (Jig Iml)

* * ____ i 1

Figure 3. Hista mine release as a function of A23 187 concentratiQn in 17 norm al donors (opell circle) and 21 patients with atopic dermatit is (closed circle) . Values are expressed as mea n :±: SEM. *Significantly difTe rcnt (rom the corresponding concentration of A23 187 of the group of control sub­jects (p < 0.05).

22 MARONE ET AL

w lOa Vl 0 u .-. <l: 0

00 °0

w • , . --' • w 80 0 • a::

0 •• w ..... 0 0 8 zoo ~~ 60

0 ;:!<l:

0 • Vl

I~ 40 ~~ ~ 20 ::>

® @ ~ x <l: a ~ a 20 40 60 80 a 20 40 60 80 lOa

MAXIMUM'!. HISTAMINE RELEASE WITH I-met peptide

Figure 4. Correlation between the maximum percent histamine release induced by the f-met-peptide and A23187 in 17 normal donors (A) and 21 patients with atopic dermatitis (B). Each symbol represents the mean of duplicate determinations. The Sperman Rank coefficients were 1',

0.52 (A) and Y, = 0.49 (B) and were significant (p < 0.05).

We have previously shown that basophil releasability induced by goat anti-IgE selectively increases with age [15] . The present results confirm that the percent of histamine release induced by rabbit anti-IgE in young normal donors is minimal. In contrast, basophils from age-matched AD pati ents show an increased his­tamine release w hen challenged w ith all anti-IgE concentrations used in this investigation. Conroy et al [38] claim that the type of anti-IgE used as stimulus is important in determining the bi­ologic response of human basophils. Although we employed throughout the experiments the preparation of rabbit anti-IgE used in previous investigations on basophi l releasability [17], we acknowledge that until a standard stimulus is available no definite conclusions can be drawn from such studies.

Previous studies have shown increased in vitro histamine release from basophils of AD patients when challenged with anti- IgE [12] or with concanavalin A (Con-A) (1 3]. T he results of these studies are difficult to compare w ith ours for several reasons. Lebel et al (12] studied histamine release from w hole blood and not from washed leukocytes. Recent results indicate that serum factor(s) might affect basophil releasability [36]. Furthermore, they used a commercia l preparation of anti-IgE and the age of the control group was not indicated. It is well established that the age of cell donors and the type of anti-IgE serum influences basophil releasability [15,16,38]. Butler et al [1 3] showed an in­creased response to Con-A in ad ult patients with AD compared with normal subjects.

We also found that basophil releasability in response to A23187 was increased in AD patients compared with controls. In contrast, the response to the f-met-peptid e was similar in the 2 groups. Interestingly, we found a significant correlation between the max­imum percent histamine release induced by the f-met-peptide and A23187 both in controls and AD. This new finding in yo ung donors contrasts with the lack of correlation generally found in adults [1 9,39], and is yet another indication that the parameter of basophil releasability must be defined w ith respect to the age of cell donor and to each stimulus.

The release of chemica l mediators from both basophils and mast cells is probably modulated by the intracellular level of cyclic AMP (cAMP) [21,23 ,26]. It has been shown that compounds that augment the intracellular cAMP exert a greater inhibitory effect on IgE-mediated histamine release from human basophils, than on f-met-peptide- [21,40] and A23187-induced histamine secre­tion [41]. Recently it has been shown that cAMP phosphodi­esterase activity is increased in leukocytes from AD patients [42] . If cAMP metabolism is deranged also in basophils from AD pa­tients it might explain some of our results. The different modi­fications of basophil releasability in response to anti-IgE, the f-met-peptide, and compound A23187 could be explained at least in part by their different sensitivity to intracellular cAMP levels.

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The in vitro abnormalities of basophil releasability that we have found in young patients with AD might have a relevance in vivo . There is increasing evidence that IgE-mediated ac tivation of ba­sophi ls plays a pathogenic role in AD [8-10]. Furthermore, ba­sophils are the only readily accessible IgE-binding, mediator­containing cell s available at present for in vitro study. Because basophils have trigger and modulatory mechanisms similar to those of human mast cells [43], they have frequently been used to probe for pathogenic mechanisms. Finally, the correlation be­tween hi stamine release from basophils and from skin mast cells in allergic diseases [44] suggests that human basophils and skin mast cells have some similarities.

IgE- and A23187-mediated basophil releasability is genetically controlled [16,45]. AD is an allergic disease w hich runs in families [1]. T herefore the inheritance of increased basophil releasability in response to anti-IgE and A23187 might contribute to the path­ogenesis of this disorder.

Previous investigations have shown that basophil releasability is an important parameter in such allergic disorders as asthma [19,20] and chronic urticaria [1 7,18]. Interestingly in these con­ditions reduced or increased releasability was found, and this was attributed to desensitization or activation, respectively, due to in vivo exposure to releasing stimuli . Our results extend these ob­servations by showing that basophil releasability can also be en­hallCed in other allergic conditions. Therefore differences in ba­sophil releasability can be appreciated in terms of augmentation or reduction in patients w ith allergic disorders.

In conclusion, using the basophil, a readily accessible mediator­containing cell that appears to be involved in AD [8,10], we have shown that patients with AD differ from control subj ects in sev­eral parameters of releasability. Although we do not suggest that the releasing agents used in the present investigation have in vivo relevance, this analysis suggests that further studies of in vitro histamine release from AD basophils are likely to be productive.

TI,c excel/ellt techl/ica l assistoll cc of Mr . O rcsle Marino is grate}i"/)' ackllowl­edged.

REFERENCES

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2. Juhlin L, Johansson SGO, Bennich H, Hagman C, Thyresson N: Immunoglobulin E in dermatoses: levels in atopic dermatitis and urtica ria. Arch Dermatoll00:12-16, 1969

3. Ogawa M, Berger PA, Mcintyre OR, Clendenning WE, Hanover NH, Ishizaka K: IgE in atopic dermatitis. Arch Dermatol 103:575-580, 1971

4. Levi L, Meneghini CL, Rantuccio F: Activite cholinesterasique: pou­voir histaminolytique et dosage de I'histamine dans la peau de sujects sains et atteint de certaines dermatoses allergiques rapport histamine-mastocytes. Acta Allergologica 13:332-348, 1959

5. Mihm MC Jr, Soter NA, Dvorak HF, Austen KF: The structure of normal skin and the morphology of atopic eczema. J Invest Der­matol 67:305-312, 1976

6. Johnson HH , DeOreo GA, Lascheid WP, Mitchell F: Skin histamine levels in chronic atopic dermatitis. J Invest Dermatol 34:237-238, 1960

7. Juhlin L: Localiza tion and content of histamine in normal and dis­eased skin. Acta Derm Venereol (Stockh) 47:383-391, 1967

8. Henocq E, Gai llard J: Cutaneous basophil hypersensitivity in atopic dermatitis. Clin Allergy 11 :13-20, 1981

9. Mitchell EB, Crow J, Chapman MD, Jouhal SS, Pope FM, Platts­Mills T AE: Basophils in allergen-induced patch test sites in atopic dermatitis. Lancet 1:127-130, 1982

10. Mitchell EB, Crow J, Path MRC, Rowntree S, Webster ADB, Platts­Mills T AE: Cutaneous basophil hypersensitivity to inhalant aller­gens in atopic dermatitis patients: elicitation of delayed responses containing basophils fo llowing local transfer of immune serum but not IgE antibody. J Invest Dermatol 83:290-295, 1984

VOL. 87, N O. 1 JULY 1986

11. Ring J , O'Connor R: 111 vitro histamine and sero tonin release studies in atopic dermatitis. Int Arch Allergy Appllmmunol 58:322-330, 1979

12. Lebel B , Venencie PY, Sa ura t JH, Soubrane C, Paupe J: Anti-l gE induced histamine release from basophils in children with atopic dermatitis. Acta Derm Venereol (Stockh) 92:57-59, 1980

13. Butler JM , C han SC, Stevens S, H anifm JM: Increased leukocyte histamine release with elevated cyclic AM P-phosphodies terase ac­tivity in atopic dermatiti s. J Allergy C lin Immunol 71 :490-497, 1983

14. Sampson HA, Jolie PL: Increased plas ma histamine concentrations after food challenges in child ren with atopic dermati tis. N Engl J Med 311 :372-376, 1984

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