Lymphocytes and Langerhans cells in patch tests

The distribution of immunocompetent cells was analysed in allergic (nickel) and irritant (dithranol) patch tests using conventional transmission electron microscopy and labelling with monoclonal antibodies in an avidin-biotin immunoperoxidase study. The biopsies were taken 24 or 48 h after the allergen/irritant application. In allergic and irritant reactions, most inflammatory cells were OKT11 positive (pan T lymphocytes). The majority of these cells were also OKT4 positive (helper/inducer T lymphocytes), while the minority were OKT8 positive (suppressor/cytotoxic T lymphocytes). NK9 positive cells (natural killer cells) were observed in small numbers. The number of dendritic OKT6 and OKIal positive cells (Langerhans cells) in the epidermis was unaffected in allergic reactions. In irritant reactions, a normal number of OKT6 positive Langerhans cells was observed, while the number of OKIal positive cells had increased in the epidermis. Dithranol caused prominent fine structural changes in the mitochondria of the Langerhans cells, while the keratinocytes appeared largely unaffected. The present study indicates that allergic and irritant patch tests cannot be differentiated reliably using current immunohistopathological or electron microscopic techniques, in spite of the small differences observed.     

In situ immunological characterization of the infiltrating cells in positive patch tests

Skin biopsies from positive allergic patch tests were analysed by immunoenzymatic labelling of frozen sections with monoclonal antibodies. In seventeen patients the cellular infiltrate consisted of T cells admixed with Langerhans cells/indeterminate cells, but in two patients there were also many B lymphocytes. The B cells were accompanied by dendritic reticulum cells forming B-cell follicles, indistinguishable from those of normal and hyperplastic lymph nodes. There was no correlation between these two immunohistological staining patterns and the sensitizing antigen, the extent of local reaction or the time from epicutaneous application of allergen to examination (2 to 16 days). The ratio between T-helper and T-suppressor cells varied considerably, and showed no correlation with these variables. In all patients the infiltrating T cells expressed HLA-DR antigen. Transferrin receptors were identified on the infiltrating T cells in biopsies from nine patients. These data indicate activation of T cells in the infiltrate from positive patch tests, and support the functional significance of Langerhans cells in the initiation and maintenance of cutaneous contact allergy. An involvement of B cells and B-cell accessory cells in the pathogenesis of contact allergic reactions is also suggested. The presence of dendritic reticulum cells in skin infiltrates from positive patch tests may reflect a functional implication of the skin in the development of B-cell memory.     

Immunophenotyping of the eczematous flare-up reaction in a nickel-sensitive subject

An eczematous flare-up reaction, occurring at a previously involved site, which followed oral challenge with 5.6 mg of nickel in a 29-year-old nickel-sensitive woman, was biopsied and studied by immunohistochemistry. The cellular infiltrate in the dermis and epidermis at 8 days was predominantly of Leu 3a phenotype (helper/inducer T lymphocytes), with smaller numbers of Leu-2a-reactive (suppressor/cytotoxic) T lymphocytes. Many infiltrating cells were DR-positive. No increase in epidermal Leu-6-positive Langerhans cells was seen but Leu-6-reactive cells were noted in the dermal infiltrate. Keratinocytes showed some expression of class II antigen (mainly DR). In comparison with the 48-hour allergic patch test reaction, the eczematous flare-up site showed no increase in epidermal Langerhans cell numbers nor infiltration with macrophages, but the responses were similar since both showed a superficial T cell reaction in the skin.     

Lymphocyte subsets and Langerhans cells in allergic and irritant patch test reactions: histometric studies

This study has attempted to distinguish between allergic and irritant reactions to patch tests by semiquantitative histological methods. The extent of perivascular chronic inflammatory infiltrate at 72 h in irritant patch test reactions to sodium lauryl sulphate was shown to be small and very consistent, whereas in allergic reactions to nickel sulphate it was generally larger and more variable in size (p less than 0.02). The two major lymphocyte subsets (T4 and T8) were randomly intermixed in both types of reaction and formed the major component of both the perivascular and diffuse dermal infiltrate, without any evidence of selective migration. The T4:T8 ratios were similar in focal and diffuse infiltrates. The number of T6 dendritic (putative Langerhans) cells in the epidermis (per mm inner epidermal length) was usually greatly reduced in irritant reactions (5-16 mm-1, mean 10 mm-1) but remained within normal limits in allergic reactions (6-33 mm-1, mean 21 mm-1) (p less than 0.001). Comparable results were seen with other irritants (mercuric chloride and benzalkonium chloride) and other allergens (neomycin sulphate, ethylene diamine and potassium dichromate). In additional experiments, pairs of biopsies were taken from the reaction and from adjacent unaffected skin. The T6 cell density in the epidermis did not significantly differ between allergic reactions and control skin. By contrast, the irritant reactions had fewer T6 cells than the control skin (p less than 0.001).     

Fc epsilon R11/CD23 receptor distribution in patch test reactions to aeroallergens in atopic dermatitis.

There is increasing evidence that exposure to organic allergens may induce or exacerbate lesional skin in patients with atopic dermatitis. In this study, patients with atopic dermatitis were patch tested to 11 common organic allergens and to control chambers containing 0.4% phenol and 50% glycerin in 0.9% saline. In biopsies from positive patch test reactions, patch test control skin, lesional eczematous and non-lesional skin from atopic individuals, and normal skin from non-atopic volunteers, the presence and distribution of macrophages (RFD7+), dendritic cells (RFD1+), and Langerhans cells, and the expression of the low-affinity receptor for IgE (CD23) were investigated. In patch test reactions and lesional skin samples, inflammatory infiltrates of diffusely distributed macrophages (RFD7+), dendritic cells (RFD1+), T lymphocytes (RFTmix+), and Langerhans cells (CD1+) were seen, the latter being present in both the epidermis and the dermis. The numbers of Langerhans cells were reduced in the epidermis and increased in the dermis in patch test reactions and lesional skin compared to their controls. Double staining revealed a change in the distribution of CD23 antigen. In patch test control and non-lesional biopsies many macrophages and only a few Langerhans cells within the dermal infiltrates expressed this antigen. In patch test reaction and lesional skin samples, however, the proportion of CD23+ dermal Langerhans cells had increased compared to macrophages. Furthermore, in these latter samples an increased proportion of dermal CD1+ cells expressed the dendritic cell (RFD1+) marker. These results show that following antigen challenge there are marked similarities between the phenotype of the cellular infiltrate in patch test reaction and lesional skin biopsies, and also demonstrate a changing distribution of CD23 on antigen-presenting cells.     

Cytochemical and Ultrastructural Studies of the Langerhans' Cells

Abstract: In the guinea pig, experimental allergic contact dermatitis (ACD) And primary irritant contact dermatitis (PICD) were induced with different concentrations of dinitrochlorobenzene (DNCB). The epidermal Langerhans' cells (LCs) were observed sequentially by both adenosine triphosphatase (ATPase) and electron microscopy. Light microscopically, in ACD, the density and dendritic processes of LC decreased markedly within 12 h after antigen challenge. Almost no recognization LCs could be seen within 2 to 5 days. Later, LCs began to repopulale in the epidermis. Within 14 days, the density and shape of the LCs returned to normal. On the contrary, LCs changed more rapidly in PICD. The dendritic processes of LC decreased within 2 h and cell density decreased dramatically within 6 h after DNCB application. LCs also repopulated more rapidly in the epidermis. Electron microscopically, in ACD, we observed that lymphocyte-like cells apposed to LCs; LCs were activated and damaged; however, in PICD, we found neither the apposition of lymphocyte-like cells to LCs, nor the activation of LCs. LCs play an important role in the convalescence phase as well as in the early and later phases of contact allergic reaction.     

Cytochemical and Ultrastructural Studies of the Langerhans'' cells

In the guinea pig, experimental allergic contact dermatitis (ACD) and primary irritant contact dermatitis (PICD) were induced with different concentrations of dinitrochlorobenzene (DNCB). The epidermal Langerhans' cells (LCs) were observed sequentially by both adenosine triphosphatase (ATPase) and electron microscopy. Light microscopically, in ACD, the density and dendritic processes of LC decreased markedly within 12 h after antigen challenge. Almost no recognization LCs could be seen within 2 to 5 days. Later, LCs began to repopulate in the epidermis. Within 14 days, the density and shape of the LCs returned to normal. On the contrary, LCs changed more rapidly in PICD. The dendritic processes of LC decreased within 2 h and cell density decreased dramatically within 6 h after DNCB application. LCs also repopulated more rapidly in the epidermis. Electron microscopically, in ACD, we observed that lymphocyte-like cells apposed to LCs; LCs were activated and damaged; however, in PICD, we found neither the apposition of lymphocyte-like cells to LCs, nor the activation of LCs. LCs play an important role in the convalescence phase as well as in the early and later phases of contact allergic reaction.     

Cutaneous dermal Ia+ cells are capable of initiating delayed type hypersensitivity responses

The presence of Langerhans cells (LC) within the epidermis has been shown to be critical for inducing T-cell-mediated immune responses in the skin. The purpose of this study was to assess whether cells in the dermis can initiate T-cell-mediated delayed-type hypersensitivity responses in vivo. Initially, back skins from C3H mice were trypsinized to remove the epidermis. The dermis was enzymatically dispersed and filtered to obtain a cell suspension. However, dermal cells exposed to trypsin were contaminated with numerous disaggregated hair follicles. These hair follicles contained Ia+ cells (presumably LC), and upon haptenation in vitro with trinitrophenyl, initiated contact hypersensitivity reactions in vivo. We therefore used dispase in place of trypsin to prevent follicular disaggregation and to allow preparation of dermal cell suspensions free of hair follicles. These hair follicle-free dermal cells were haptenated with trinitrophenyl and injected intradermally. Elicitation of contact hypersensitivity by epicutaneous painting 6 d later revealed the mean +/- SEM incremental ear-swelling response to be 53 +/- 8 mm X 10(-3). In contrast, mice sensitized by injection with dermal cells depleted of Ia+ cells demonstrated only 10 +/- 1 mm X 10(-3) of ear swelling. Thus, like dendritic LC of the epidermis, perivascular dendritic Ia+ cells of the dermis are capable of initiating T-cell-mediated contact hypersensitivity in vivo and may be highly relevant for presentation of antigen to T cells trafficking through the dermis.     

Flow cytometrically-sorted residual HLA-DR+T6+ Langerhans cells in topical steroid-treated human skin express normal amounts of HLA-DR and CD1a/T6 antigens and exhibit normal alloantigen-presenting capacity

Topical corticosteroids decrease the number of HLA-DR+T6+ Langerhans cells (LCs) and the antigen-presenting capacity of epidermal cells (ECs). We have investigated the properties of residual HLA-DR+T6+ LCs in steroid-treated human skin. Flow cytometric analysis revealed that clobetasol propionate 0.05% applied twice daily for 7 d reduced the percentage of HLA-DR+T6+ LCs in EC suspensions to 46% of control (from a mean percentage +/- sem of 2.49 +/- 0.30 in control skin to 1.15 +/- 0.22 in steroid-treated skin), but did not significantly alter the relative amounts of HLA-DR and CD1a/T6 antigens per individual HLA-DR+T6+ cell. HLA-DR+T6- and HLA-DR-T6+ cells were not detected in either group. Steroid therapy significantly decreased the allostimulatory capacity of unsorted ECs. By contrast, in parallel experiments in which the same EC suspensions were greatly enriched (85% to 90%) for HLA-DR+T6+ LCs by flow cytometric sorting, the allostimulatory capacity of purified LCs from steroid-treated skin was not significantly different from control. Residual HLA-DR+T6+ LCs, which preserve their antigenic markers and alloantigen-presenting function, may be relatively unaffected because they have only recently immigrated into the epidermis, or they may represent a subgroup of steroid-resistant LCs. Alternatively, given the dose response relationship between topical steroid potency and decrease in HLA-DR+T6+ LC numbers, the apparent steroid resistance of residual HLA-DR+T6+ LCs may reflect heterogenity in the density of expression of LC steroid receptors.     

Expression of cyclooxygenase isozymes during morphogenesis and cycling of pelage hair follicles in mouse skin: precocious onset of the first catagen phase and alopecia upon cyclooxygenase-2 overexpression

Cyclooxygenase (COX)-1 and -2 catalyze the key reaction in prostaglandin biosynthesis. Whereas COX-1 is found in most tissues, COX-2, with a few exceptions, is not expressed in normal tissues but becomes transiently induced in the course of inflammatory reactions. In many neoplastic epithelia, COX-2 is constitutively overexpressed. Here we show that COX isozymes are spatiotemporally expressed during morphogenesis of dorsal skin epithelium of NMRI mice. COX-1 and COX-2 mRNA and protein were detected in embryonic and postnatal epidermal tissue by RT-PCR, northern blot, and immunoblot analysis indicating that both isoforms may contribute to prostaglandin production. Being barely detectable in interfollicular epidermis and resting hair follicles of adult mice, COX-2 protein appeared in embryonic skin first in epidermal precursor cells and later on in the basal cells and the peridermal layer of the stratified epidermis. In the course of pelage hair follicle morphogenesis, COX-2 remained expressed in the basal interfollicular compartment and, in addition, became apparent in elongated hair germs and hair pegs and later on in the outer root sheath cells of the distal and proximal hair follicles as well as in basal sebaceous gland cells. During the subsequent synchronous phases of hair cycling, COX-2 expression declined in catagen, was barely detectable in telogen, and was reinduced in the basal outer root sheath and basal sebaceous gland cells of anagen hair follicles. COX-1 immunosignals were detected predominantly in the interfollicular spinous and granular layers of the developing, neonatal, and adult epidermis but not in follicular epithelial cells of developing or cycling hair follicles. Dendritic cells in the interfollicular epidermis and distal hair follicles were also COX-1-positive. Transgenic overexpression of COX-2 under the control of a keratin 5 promoter in basal cells of the interfollicular and follicular epidermis induced a precocious entry into the first catagen stage of postnatal hair follicle cycling and a subsequent disturbance of hair follicle phasing. Furthermore, transgenic mice developed an alopecia. Inhibition of transgenic COX-2 activity by feeding the specific COX-2 inhibitor valdecoxib suppressed the development of alopecia, indicating that COX-2-mediated prostaglandin synthesis is involved in hair follicle biology.     

Effect of cignolin and infrared irradiation on the patch test and lymphocyte transformation test

In a study on the effect of anthralin and infrared irradiation (IR) on the allergic patch test in vivo and the lymphocyte transformation test in vitro, we observed that anthralin enhanced the local test reaction. Our findings suggest an additive reaction of toxic anthralin dermatitis and allergic test reaction. Immunohistology showed that additional treatment with anthralin resulted in elevated numbers of the OKT-6+ dendritic cells in the epidermis. Anthralin in concentrations of greater than or equal to 10(-5) M inhibited the lymphocyte transformation in vitro. IR irradiation-either before or during patch testing-did not significantly influence the allergic test reaction or the lymphocyte transformation, if the temperature was adjusted to 37 degrees C. In comparison to convective heat, we found no specific effect of IR irradiation.     

Long-lasting allergic patch test reactions to nickel sulfate: analysis by nickel quantification and immunocytochemistry

We previously showed the median duration of positive patch test reactions to nickel sulfate(5% pet) was 9 days, and defined as long-lasting (LLAPTR) the 14.3% of reactions that persisted for 17 Days or longer. The pathomechanisms of LLAPTR are unclear, but may involve either localized antigen persistence or abnormal down regulation of the cellular immune response. In this study, we compared (a) the nickel concentration and (b) the immunocytochemical nature of the local immune reaction, between biopsies from LLAPTR ( n = 8) and normally resolving allergic patch lest reactions (NRAPTR) ( n = 8) to nickel sulfate. The concentration of nickel in LLAPTR (median 0.8μg/g, μg/g, range 0.25–3.87 μg/g, mean 0.83μg/g, 95% CI 0 35–1.31) and NRAPTR (median 0.58 μg/g, range 0.2 1.85 μg/g, mean 0.88 μg/g, 95% CI 0.02 1.74) was similar. Activated T lymphocytes, expressing surface IL-2 receptor, HLA DR, DR alpha 1, DP, DQ, and CD2>CD8>CD4 antigens, were seen throughout the dermis and occasionally infiltrating the suprabasal layer of the epidermis in all biopsies. CDI and HLA DR, DR alpha 1, DP, and DQ-expressing Langerhans cells were present throughout the epidermis and occasionally seen in the papillary dermis. HLA DR, DR alpha 1, DP, and DQ antigen expression were also seen on the surface of non-dendritic cells in the epidermis (probably either keratinocytes or T lymphocytes) and vascular endothelial cells in the papillary dermis. There were no significant qualitative or quantitative differences in the immuno-cytochemical nature of the localized immune reaction between LLAPTR and NRAPTR. These findings suggest that the pathomechanism of LLAPTR to nickel sulfate is unlikely to be explained simply on the basis of nickel concentration or the nature of the localized immune reaction at the patch test site.     

Defined in situ enumeration of T6 and HLA-DR expressing epidermal Langerhans cells: morphologic and methodologic aspects

An essential prerequisite for the in situ enumeration of epidermal Langerhans cells (LCs) is the unequivocal identification of the desired cell type. We have examined over 250 cryostat sections of normal human skin to analyze morphologic and methodologic problems underlying the quantification of epidermal LCs, defined by anti-T6 (OKT6) and anti-HLA-DR (OKIal) immunoperoxidase staining. Our findings show that OKT6 reactivity of dendritic processes in cross-sectioned epidermis yields microscopic images which are not easy to analyze objectively. The morphology that we find leads us to categorize dendritic cells into 3 arbitrary types of T6+ LC profiles. In addition we describe criteria for the assessment of OKT6 staining patterns relating to the dendritic state of epidermal LCs. Preliminary quantitative data on this issue are discussed in relation to: epidermal thickness; the thickness of skin tissue sections; and the discrepancy between the number of T6+ and HLA-DR+ LCs. We hope that the principles outlined in this report may serve to overcome potential methodologic problems with quantitation of T6+ epidermal LCs in skin sections.     

The purpuric patch test in patients with allergic contact dermatitis from azo dyes

The histopathological features of the purpuric patch test have been described in individual cases only. We report a series of patients with allergic contact dermatitis, who developed purpuric patch tests at the sites of allergens from the azo dye group. 105 patients were clinically evaluated and tested with the TRUE Test and the textile color & finish series (Chemotechnique Diagnostics) because of suspected clothing dermatitis. Positive results to the latter were found in 31 patients (29.5%). In 9 of these, purpuric patch tests were observed at the sites of the allergens Disperse Blue 124, 106 and 85. 10 biopsies were performed and studied. The histopathological changes of the purpuric patch test included: spongiosis (in 90% of cases), exocytosis (70%), and dilated blood vessels (100%) without signs of vasculitis, surrounded by an inflammatory infiltrate composed mainly of T lymphocytes. Extravasated erythrocytes were seen perivascularly, but also in the interstitium, surrounding the acrosyringium, at the dermoepidermal junction, and in the epidermis. Increased number of mast cells were found in 22.2% of cases. Disperse Blue 124, 106, and 85 are potent allergens that can elicit purpuric patch test reactions. The purpuric patch test in our cases was a manifestation of an allergic reaction, based not only on histopathological changes, but also on evolution and relevance of the patch tests.     

Abnormal expression of class I and class II major histocompatibility antigens in alopecia areata: modulation by topical immunotherapy

Fifty-eight scalp biopsies were immunohistologically investigated with monoclonal antibodies against HLA-ABC, HLA-DR, and T6 antigens. The following 3 groups were compared: control biopsies obtained from healthy volunteers (n = 5) or patients with unrelated scalp diseases (n = 6); biopsies from untreated alopecia areata (AA), obtained either from untreated patients (n = 19) or from the untreated side in patients receiving unilateral treatment with the contact allergen diphencyprone (DCP) (n = 13); biopsies obtained from the treated side in patients receiving unilateral treatment with DCP (n = 13). While HLA-ABC antigens were strongly expressed by epidermal keratinocytes and the infundibular epithelium of hair follicles in all biopsies, these antigens were either not detectable or only faintly expressed on the subinfundibular epithelium and the hair matrix in the control series. By contrast, 30 out of 32 biopsies from untreated AA showed expression of HLA-ABC antigens on hair matrix epithelium, and the subinfundibular epithelium was HLA-ABC-positive in 15 out of 32 cases. In the biopsies from treated AA, HLA-ABC antigens were expressed on hair matrix epithelium in 9 out of 13 cases, and on the subinfundibular epithelium in 1 case. In the controls and untreated AA, HLA-DR expression was confined to dendritic cells in the epidermis and the follicular infundibulum. Its expression on hair matrix epithelium was found in 15 out of 32 biopsies from untreated AA and in 4 out of 13 biopsies from treated AA. In the control series, intrabulbar T6+ dendritic cells were either absent or present in low numbers. High numbers of intrabulbar T6+ cells were present in 7 out of 32 biopsies from untreated AA and in 0 out of 13 biopsies from treated AA. The data show that abnormal expression of class I major histocompatibility (MHC) antigens on hair matrix epithelium is a constant feature in AA, whereas class II MHC antigens are less frequently expressed. Topical immunotherapy with DCP, which induced expression of HLA-DR in epidermal keratinocytes in 6 out of 13 cases, reduced the abnormal expression of both HLA-ABC and -DR antigens in the epithelium of lower hair follicles in AA.     

Result of "as is patch test" using mite elements in atopic dermatitis (AD) patient. 2nd report, "Mite lipid"

The antigenicity of mite lipid was studied, taking atopic dermatitis as an allergic disease. Lipid was extracted from Dermatophagoides pteronyssinus (Dp) and D. farinae (Df). It was tentatively designated a mite lipid antigen (Dp-L, Df-L), and then a patch test was performed. An extract from animal food (LC) and acetone were used as the controls. With only Dp-L, a positive reaction of more than one plus (+) of the ICDRG standard was noted in 3 out of 21 cases of AD. Histology of the above-mentioned positive cases revealed spongiosis and infiltration of small round cells in the upper dermis along with scattered eosinophiles. Leu 6 positive cells were noted frequently in the epidermis and dermis. The lipid components of Dp-L were made up mostly of phospholipid and triglycerides+, and lipoprotein was not contained. Judging from the findings above it appears necessary to study not only protein but also lipid in connection with the mite antigenicity++ in AD in the future.     

Mite-antigen-stimulated cytokine production by peripheral blood mononuclear cells of atopic dermatitis patients with positive mite patch tests

To investigate possible involvement of Th1-type immunoreaction in the development of skin lesions of atopic dermatitis (AD), we measured mite-antigen-stimulated production of IL-2, IL-4 and IFN-γ, using peripheral blood mononuclear cells (PBMC) obtained from 23 patients with AD who developed positive patch test reactions to house dust mite antigens extracted from Dermatophagoides pteronyssinus (Dp). Incubation of these PBMC with the Dp antigen for 72 h produced marked secretion of IL-2 and IFN-γ, but not IL-4, indicating that Dp-specific T cells were found in these patients' circulating peripheral blood and were capable of producing IL-2 and IFN-γ, known as major mediators of the delayed-type allergic reaction. Our study suggests that Thl-type cells may be involved in the development of skin lesions of AD patients with positive patch test reactions to house dust mite antigen.     

小鼠毛囊周期中Thy-1抗原的表达

目的　观察拔毛诱导的毛发生长周期中毛囊Thy 1抗原的表达。 方法　自然休止期C57BL/6小鼠 ,通过拔毛诱导其毛发自休止期进入生长期 ,应用ABC免疫组化法连续观察毛囊Thy 1抗原的表达及组织学改变。结果　拔毛后第 2天 ,漏斗部毛囊内层角质形成细胞明显表达Thy 1抗原 ,随后转为毛囊周围浸润细胞及基质较强的表达Thy 1抗原 ,以 7～ 9天最强 ,以后逐渐减弱 ,至 17～ 2 0天基本消退。结论　拔毛刺激毛发生长周期中毛囊上皮可一过性表达Thy 1抗原。毛囊上皮的这种Thy 1抗原的表达 ,在皮肤免疫应答中可能起重要的作用     

Allergic contact dermatitis due to phenylephrine hydrochloride, with an unusual patch test reaction

2-day (2-D) closed patch tests are often used in daily clinical practice and useful for evaluating the cause of allergic contact dermatitis. However, even when 2-D closed patch tests at appropriate concentrations are performed for suspected allergic contact dermatitis based on clinical findings, positive reactions are not always obtained. Therefore, although the use of the allergen again induces similar symptoms, a definite diagnosis cannot be made in some cases. We report a case of allergic contact dermatitis due to phenylephrine hydrochloride in eyedrops, with an unusual patch test reaction. Although the results of the routine 2-D closed patch test were negative, a definite diagnosis could be made by closed scratch-patch test. In addition, long-lasting allergic patch test reactions were observed at the positive scratch-patch test site for about 3 months. We speculated that these unusual results on patch testing in our case were associated with the degree of percutaneous absorption of causative agents. Therefore, even when 2-D closed patch tests are negative, scratch-patch tests may be indicated for patients in whom clinical symptoms continue strongly to suggest contact dermatitis.     

The duration of Grenz ray-induced suppression of allergic contact dermatitis and its correlation with the density of Langerhans cells in human epidermis

Recent investigations have shown that Grenz rays can suppress the allergic contact dermatitis reaction completely and that Langerhans cells, identified by OKT6 antibodies and electron microscopy, disappear from the epidermis at the same time. It is not known for how long this suppression lasts. This has been investigated in 28 nickel-sensitive patients who were given Grenz rays (3 Gy) on the back, once a week for 3 weeks. The patients were then divided into four groups and tested with patch tests for nickel at 1, 7, 14 and 21 days after the last Grenz ray treatment. Biopsies were taken from positive patch test sites, and from the corresponding opposite control. They were labelled with OKT6 antibodies to detect Langerhans cells. The patch test reactions were suppressed and the Langerhans cell density was decreased initially. These changes were restored after 3 and 6 weeks, respectively. The results show that the effect of Grenz rays on eczematous reactions extends to a maximum of 3 weeks and imply that Langerhans cells are necessary for the elicitation of the efferent phase of allergic contact dermatitis.