FceRll/CD23 Receptor Distribution in Patch Test Reactions to Aeroallergens in Atopic Dermatitis Colin C. Buckley, Carol Ivison, Leonard W. Poulter, and Malcolm H.A. Rustin Departments of Dermatology and Immunology (LWP), The Royal Free Hospital and School of Medicine, London, U.K. There is increasing evidence that exposure to organic aller­ The numbers of Langer hans cells were reduced in the epider­ gens may induce or exacerbate lesional skin in patients with mis :and increased in the dermis in patch test reactions and atopic dermatitis. In this study, patients with atopic dermati­ lesional skin compared to their controls. Double staining tis were patch tested to 11 common organic allergens and to reve:aled a change in the distribution of CD23 antigen. In control chambers containing 0.4% phenol and 50% glycerin patch test control and non-Iesional biopsies many macro­ in 0.9% saline. In biopsies from positive patch test reactions, phages and only a few Langerhans cells within the dermal patch test control skin, lesional eczematous and non-lesional infiltrates expressed this antigen. In patch test reaction and skin from atopic individuals, and normal skin from non­ lesional skin samples, however, the proportion of CD23+ atopic volunteers, the presence and distribution of macro­ dermal Langerhans cells had increased compared to macro­ phages (RFD7+), dendritic cells (RFD1 +), and Langerhans phages. Furthermore, in these latter samples an increased cells, and the expression of the low-affinity receptor for IgE proportion of dermal CD 1+ cells expressed the dendritic cell (CD23) were investigated. (RFD 1+) marker. These results show that following antigen In patch test reactions and lesional skin samples, inflamma­ challenge there are marked similarities between the pheno­ tory infiltrates of diffusely distributed macrophages type of the cellular infiltrate in patch test reaction and le­ (RFD7+), dendritic cells (RFD1+), T lymphocytes sion:al skin biopsies, and also demonstrate a changing distri­ (RFTmix+), and Langerhans cells (CD1+) were seen, the bution of CD23 on antigen-presenting cells.] Invest Dermatol latter being present in both the epidermis and the dermis. 99:184-188,1992 or many years the role of IgE in the pathogenesis of It has been shown that AD can relapse following environmental atopic dermatitis (AD) has been controversial. Approxi­ Contact with specific allergens, and the dermatosis remits if these mately 80% of patients with AD show an immediate allergens are removed from the patient'S environment [6,10]. The type hypersensitivity reaction to environmental aller­ aeroallergens in these and other studies [5,7,8,11,12] were identi­ gens when given intradermally as a skin prick test, to­ fied by their ability to induce both positive skin prick test reactivity getFher with elevated serum IgE levels with a specificity to these and positive patch test reactions. Delayed patch test reactions oc­ allergens [1] . Despite this, the cellular characteristics of the eczema­ curred only to allergens to which the patient reacted when chal­ tous lesion are repeatedly described as that of a delayed-type hyper­ lenged in the skin prick test. Significantly, these reactions uni­ sensitivity reaction, with the T lymphocyte as the predominant cell formly failed to exhibit the clinical or histologic features of an type in the dermal infiltrate [2 - 4] . Epicutaneous patch testing with immediate hypersensitivity reaction when analyzed at 48 h. It has ae roallergens such as house dust mite (Dermatophagoides pterol1Ys­ therefore been suggested that the skin lesions of AD result from a sinus) produces an eczematous reaction, which also has the dy­ chronic delayed hypersensitivity reaction involving epidermal Lan­ namics and histology of a delayed-type hypersensitivity reaction gerhans cells that present antigen to sensitized T lymphocytes [13]. [5-9]. Though the positive patch test reaction and lesional skin in pa­ tients with AD do not exhibit the clinical or histologic features of a type 1 hypersensitivity reaction, IgE may playa role in presenting aeroallergens. In a recent in vitro study, cell-bound IgE on Langer­ Manuscript received July 9, 1991; accepted for publication February 27, hans cells was required to initiate T-cell responses to allergens [14]. 1992. Further work has shown that in patients with AD the ratio of This resea rch was supported by grants from The Sir Jules Thorn Charita­ CD 1- + /IgE+ cells in the epidermis decreases after 1 week of treat­ ble Trust and The Skin Disease Research Fund. ment with topical corticosteroid, with complete disappearance of Part of this work was presented at the 52nd annual meeting of the Sociery IgE+ cells after 2 weeks [15]. Low-affinity receptors for IgE have for Investigative Dermatology, Seattle, May 1991. been identified on epidermal Langerhans cells as well as on macro­ Reprint requests to: Dr. C. Buckley, Department of Dermatology, The Royal Free Hospital, Pond Street, Hampstead, London NW3 2QG, En­ phages, Band T lymphocytes, and platelets [16] . This has been gland. observed in biopsies both from patients with AD and non-atopic Abbreviations: individuals [17]. Although the precise function of these receptors in AD: atopic dermatitis AD is unclear, it could be hypothesized that they may playa role in APAAP: alkaline phosphatase - anti alkaline phosphatase the presentation of antigen to activated T lymphocytes. In this FlTC: fluorescein isothiocyanate study, we have investigated CD23 expression on Langerhans cells, MoAb: monoclonal antibody dendritic cells (RFDl +), and macro phages (RFD7+) in both posi­ RF: The Royal Free Hospital School of Medicine ti"e patch test reactions and non-patch tested lesional skin from TBS: Tris-buffcred saline patients with AD. We also wished to study the relative distribution TRITC: tetramethylrhodamine isothiocyanate 0022-202X/92/S05.00 Copyright © 1992 by The sociery for Investigative Dermatology, Inc. 184 VOL. 99, NO.2 AUGUST 1992 CD23 RECEPTORS IN ATOPI C DERMATITIS 185 of these subsets of non-lymphoid immunocompetent cells in the Table I. List of Monoclonal Antibodies Used in the Study positive patch test reaction, patch test control, lesional, and non-le­ Reactivity in sional biopsies. Name CD Subclass Source Normal Tissues MATERIALS AND METHODS RFDl IgMp RFHSM' Epitope of HLA-DR Patients Fifty-six patients (37 male, 19 female) with atopic der­ with restricted ex- matitis (age range, 22 - 38 years) diagnosed according to the criteria pression to dendritic of Hanifin and Rajka [18] were studied. Patients had moderate to cells severe dermatitis requiring daily topical corticosteroids. Seven of RFD7 IgGl RFHSM Mature tissue macro- the patients had bronchial as thma and 20 suffered from seasonal phages RFT6 CDl IgGy RFHSM Thymocytes and Lan- allergic rhinitis. .. gerhans cells All patients were skl11 prick test~d on the volar aspect of the RFTmix CDS,8, IgGy RFHSM Pan T lymphocyte non-dominant forearm to the followl11g common mhaled allergens: CDll Cladosporium herbartlm, Alternaria telnlis, Candida albican s, Dermato­ FceR11 CD23 IgG1y Dakopatts Low-affinity IgE recep- phago ides pteronyssinus, Dermatophago ides jarinae? cat and dog dander, UK Ltd. tor birch and plane tree extracts, rye grass, and ttmothy grass pollens • RFHSM, Royal Free Hospital School of Medicine. (Pharmacia Ltd.), using 1 % histamine as a positive control and 0.4% phenol and 50% glycerin in 0.9% saline as a negative contro!' Aeroallergen Patch Testing and Skin Biopsies All patients Quantification of the distribution of specific cell rypes was per­ had not applied topical corticosteroids to the areas to be patch tested formed using an image analyzer to define and measure framed areas for 2 weeks prior to testing and none had received ultra-violet 13 of section (one high-power field [X 400] below the dermo-epider­ irradiation, PUV A, or treatment with immunosuppressive agents in mal junction) to be counted and point counts made of the positively the previous month. stained cells in at least six areas. The results were expressed as the Epicutaneous patch testl11g was perfor.me? O? all pattents .wah number of cells per unit area. Counts of RFD 1+ cells were) per­ 8-mm Finn chambers (Epltest Ltd, Helsl11b, Fmland) contalmng formed using a fluorescence microscope and the results expressed as the same allergens in the same concentrations as used for skin prick the number of cells per high-power view (X 400). testing, on clinically normal skin on both sides of the upper back, one side having been prepared by Sellotape stripping 15 times. One Statistical Methods Where appropriate, statistical differences chamber on either side contained diluent as contro!' The patch tests between densities of the cellular infiltrates were determined using were examined at 48 h and the positive patch test reactions were the Wilcoxon sign rank test for paired data; a value of p < 0.05 was graded according to the International Contact Dermatitis Group considered significant. guidelines [19]. RESULTS Ten patients consented to have 4-mm .punch bIOpSies from the Sellotape-strippe~ poslt.lve pa.tch test reactlO~ and pat~h test control All patients with AD were skin prick test positive to one or more sites. For companson, SIX pattents With atopic dermatttts who were inhalant allergens. Seventeen patients showed positive patch test skin prick test positive had biop.sies tak~n from non-t a p ~ -stripped reactions to one or more epicutaneously applied allergens. The lesional eczematous and non-leslOnal skl11. Non - tape-stnpped skin commonest reactions were to Dermatophagoides pteronyssinus and biopsies from five age- and sex-matched normal individuals with nO Dermatophagoides jarinae, cat dander, Candida albicans and Cladospor­ personal or family history of atopy and who were skin prick test ium herbanlm, and rye grass pollen. T en patients reacted to more negative acted as further control samples. Specimens were placed in than one allergen and four patients developed positive patch test embedding compound (Bright Instruments Company Ltd, UK), reactions on both the Sellotape-stripped and non-stripped sides, snap frozen in liquid nitrogen, and stored at -70 ° C.