The skin in severe combined immunodeficiency: a case with transient cutaneous presence of gamma/delta (TRC1+) T cells.

We report the case of a boy with severe combined immunodeficiency (SCID) and serious skin problems. The level of purine 5'-nucleotidase was greatly reduced in the lymphocytes of this patient. To our knowledge, no patients with SCID and this enzyme deficiency have been described previously. The relationship between reduced levels of this enzyme and the immunodeficiency is unclear. This case is also unusual because of the presence of large numbers of T lymphocytes expressing TCR1 (gamma/delta) in the skin. Moreover, the presence of so many TCR1-positive cells was not consistent with the low numbers of these cells in the peripheral blood. These cells were not present in skin biopsies taken at a later stage during the course of the disease. An oligoclonal lymphocytosis developed during follow-up, and a monoclonal antibody reactive with these clones was found, indicating that these lymphocytes were present in the skin. This case report illustrates the benefit of the use of monoclonal antibodies in identifying the cells involved in the cutaneous inflammation in SCID, in order to gain a better insight into the characteristics of these cells.

Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues.

The polymerase chain reaction (PCR) based on the selective amplification of a 530-bp fragment of the gene encoding the proline-rich antigen of Mycobacterium leprae was applied on sections of fixed or frozen biopsy samples from leprosy patients. A simple procedure for the extraction of DNA from M. leprae in clinical specimens that provided suitable template DNA for amplification was developed. When PCR was applied on frozen sections, positive amplification in samples from all untreated acid-fast bacillus (AFB)-positive patients and in samples from 56% of the untreated AFB-negative patients could be detected, while biopsy samples from patients with skin diseases other than leprosy were all PCR negative. With neutral Formalin-fixed biopsy samples, positive amplification in 92% of the samples from untreated AFB-positive patients and in 61% of the samples from untreated AFB-negative patients could be detected by PCR. Biopsy samples exposed to mercuric chloride or nonbuffered formaldehyde containing fixatives were not suitable for application of PCR. This PCR holds promise as a tool for studies on M. leprae infection.

Discoid lupus erythematosus-like lesions in carriers of X-linked chronic granulomatous disease.

A questionnaire was sent to 16 carriers of the X-linked cytochrome-b558 negative variant of chronic granulomatous disease (CGD). Of the 15 who answered the questionnaire and from data of one additional case, 70% reported recurrent aphthous stomatitis and 63% had recurrent skin eruptions. Five of the carriers (31%) had clinically discoid lupus erythematosus (DLE), although the histopathology was not typical and the immunofluorescence findings were negative. The infiltrate in the lesional skin of three of these patients was analysed using monoclonal antibodies against CD3, CD4, CD8, CDIa, HLA-DR, CDIc, interdigitating cells, B lymphocytes and cells of the monocyte/macrophage lineage. The immunophenotype of the infiltrating cells resembled that found in discoid lupus erythematosus.

Human epidermal Langerhans cells undergo profound morphologic and phenotypical changes during in vitro culture.

Morphology, phenotype, and enzyme activity of highly enriched (80%) unlabeled human epidermal Langerhans cells (LC) have been studied, with emphasis on changes during a short-term culture of three days in vitro. All freshly isolated LC contained Birbeck granules and expressed high levels of CD1a, CD1c, and MHC class II molecules HLA-DR, -DP, and -DQ. They have a weak to moderate expression of RFD1, C3biR, Fc gamma R, p 150/95, MHC class I molecules HLA-ABC, and of the adhesion molecules LFA-3 and ICAM-1, whereas no expression of LFA-1 and several monocyte/macrophage markers were detected. Human LC undergo profound changes during in vitro culture. Birbeck granules, C3biR, Fc gamma R, and p 150/95 were completely lost and the expression of CD1a and CD1c was markedly decreased or lost. Expression of molecules that have essential functions in antigen presentation remained present at the same level (MHC class II molecules and ICAM-1) or was markedly enhanced (LFA-3 and MHC class I). Highly remarkable was the dramatically enhanced expression of interdigitating cell marker RFD1. The monocyte/macrophage markers initially absent remained absent and the enzyme activity initially present (including ATPase and nonspecific esterase) remained present. In conclusion, the results in this report stress rapid alterations of human LC during in vitro culture, resulting in transformation into cells that have phenotypical characteristics of potent antigen presenting cells that resemble interdigitating cells.

T-cell receptor gamma delta bearing cells in normal human skin.

T-cell antigen receptors (TCR) are divided into common alpha beta and less common gamma delta types. In the murine skin, TCR gamma delta+ cells have been reported to form the great majority of epidermal T lymphocytes. We have examined the relative contribution of TCR alpha beta+ and TCR gamma delta+ cells to the T-cell population in normal human skin. Serial sections of freshly frozen skin specimens were acetone fixed, incubated with anti-CD3, beta F1 (anti-TCR alpha beta), anti-TCR gamma delta-1 and anti-TCR delta 1 (anti-TCR gamma delta) monoclonal antibodies (MoAb), and stained with a highly sensitive method. Over 90% of the T cells of normal human skin are localized around the postcapillary venules of the dermis, while less than 5% are present within the epidermis. In papillary dermis, TCR gamma delta+ cells formed on average 7% (anti-TCR gamma delta-1) or 9% (anti-TCR delta 1) of the total number of CD3+ cells, while TCR alpha beta+ cells constituted up to 80%. In epidermis, these percentages were 18% and 29% for TCR gamma delta+ cells, and up to 60% for TCR alpha beta+ cells. It is concluded that there is no preferential immigration or in situ expansion of TCR gamma delta+ T cells in normal human skin, because the relative percentages found for the TCR alpha beta+ and TCR gamma delta+ populations in skin are comparable to those found in lymphoid organs and peripheral blood. However, the percentage of TCR gamma delta+ cells in epidermis seemed on average higher than in papillary dermis. Therefore, there may still be a difference in migration patterns of TCR gamma delta+ v TCR alpha beta+ cells, but this does not result in their preferential localization in human epidermis. The hypothesis that TCR gamma delta+ T cells have a specialized function in immunosurveillance of epithelia may thus not be valid for human epidermis.

Quantification and distribution of lymphocyte subsets and Langerhans cells in normal human oral mucosa and skin.

Normal human oral (check) mucosa was studied to discover whether the oral cavity resembles the Mucosal Immune System (MIS) or the Skin Immune System (SIS). Immunophenotypes of lymphocyte subsets and Langerhans cells (LC) with their exact locations in the epithelium and papillary layer of the normal buccal mucosa were determined and compared with data of normal human skin. In a double staining procedure, the distribution of T-lymphocytes in relation to blood and lymph vessels was determined. Immunophenotyping of LC was done with a CD1a monoclonal antibody. In contrast to the skin, T-lymphocytes in buccal mucosa are not primarily perivascular in location. They are more or less randomly distributed on both sides of the basement membrane. The epithelium of the buccal mucosa contains about 37 times as many T-lymphocytes as the epidermis of normal skin. T-cell numbers in the papillary layer are more or less comparable. The CD4/CD8 ratios of about 1/2 in the epithelium of buccal mucosa and 1/4 in the skin indicates preferential presence of the CD8 subset in both sites, but the helper/inducer T-lymphocytes play a much greater role in the epithelium of the buccal mucosa when compared with skin. B-lymphocytes were not found in the epithelium and papillary layer of the buccal mucosa. Thus, immune response associated cells in buccal mucosa do not show the MIS pattern since B cells are absent. It has more in common with SIS but differences are also apparent. In the epithelium of the buccal mucosa the density of LC does not differ significantly from that of the skin, but the papillary layer of the buccal mucosa contains significantly fewer LC than the skin. As in the skin most of the LC of the buccal mucosa are found in the epithelium.

Increased anthralin irritation response in vitiliginous skin.

The irritation response to anthralin was studied using the chamber-testing technique in 17 patients with vitiligo. Anthralin concentrations of 0.1%, 0.5%, 1%, and 5% in lanette wax were applied to both vitiliginous and adjacent pigmented skin for 24 h. The extent of the erythematous reaction was evaluated on the 2nd day after application. The visual assessment of the paired anthralin patches indicated that the erythema was more intense in pigmented skin than in vitiliginous skin in 15 out of 17 patients. Chromometer readings, however, clearly indicated that the erythematous response was stronger in the vitiliginous skin than in the pigmented skin, confirming the known fact that the human eye is not accurate in the quantitative assessment of complex colors. Immunophenotypification of cellular infiltrates, using the combination of different monoclonal antibodies and the peroxidase technique, showed that inflammatory cell infiltrates caused by the anthralin exposure contained increased numbers of granulocytes and monocytes in vitiliginous skin when compared with normal skin. The percentage of T-cell subsets, Langerhans cells, and mast cells in the same infiltrates of both types of skin were similar. Our results are discussed in accordance with the view that anthralin-induced radical species of the pigmented skin can be neutralized by the scavenging properties of melanin.

The poikiloderma of Rothmund-Thomson syndrome: changes in Langerhans cell morphology and distribution.

A black girl with the Rothmund-Thomson syndrome is presented. Immunophenotyping of subpopulations of immunocompetent cells in a biopsy of an atrophic hyperpigmented skin lesion revealed sparsity and unusual distribution of epidermal Langerhans cells. These cells were mainly located in the basal layer of the epidermis and did not show the usual dendritic pattern. Impressive immunoreactivity of the dermal infiltrate was observed by anti-HLA-DR staining. The changes in Langerhans cell morphology and distribution may indicate functional impairment of the up-regulating arm of skin immunity.

Predominance of "memory" T cells (CD4+, CDw29+) over "naive" T cells (CD4+, CD45R+) in both normal and diseased human skin.

Absolute numbers of CD3+ T lymphocytes and their subpopulations were determined and statistically evaluated in the lesional skin of psoriasis, atopic dermatitis, nummular dermatitis, pityriasis rosea, and lichen planus. Skin sections were divided into horizontal layers and the numbers of CD3+ T cells as well as CD4+ inducer and CD8+ suppressor-cytotoxic T-cell subsets were counted. In addition, absolute numbers of the two subpopulations of inducer T cells, i.e., "memory" (4B4+ 2H4-) and "naive" (4B4- 2H4+) were evaluated. Unexpectedly, epidermal infiltration by T cells was highest in psoriasis and lowest in atopic dermatitis. In most cases, this exocytosis was dominated by CD8+ suppressor/cytotoxic T lymphocytes, with a minimal epidermal mean CD4/mean CD8 ratio of 0.04 in pityriasis rosea and a maximum of 0.48 in psoriasis. Inducer T cells within the epidermis were almost exclusively of the 4B4+ 2H4- "memory" T-cell subpopulation, whereas 4B4- 2H4+ "naive" T cells were extremely uncommon in lesional epidermis. Similar results were obtained for dermal T cells in all diseases studied, i.e., 4B4- 2H4+ "naive" T cells were relatively rare. Papillary dermis infiltration by T cells was highest in lichen planus where a mean CD4/mean CD8 ratio of 1.10, the minimum in this comparative study, was obtained. The mean CD4/mean CD8 ratio of the papillary infiltrate was highest in atopic dermatitis (4.12). Our results indicate disease-specific and significantly different infiltration patterns of T-lymphocyte subsets in the chronic inflammatory dermatoses investigated.(ABSTRACT TRUNCATED AT 250 WORDS)

T-lymphocyte and Langerhans cell distribution in normal and allergically induced oral mucosa in contact with nickel-containing dental alloys.

An in vivo comparison was made between the contact allergic stomatitis-inducing capacity of nickel, nickel-containing dental alloys and a non-corrosive precious metal. Fifteen patients with a positive allergic skin reaction to nickel were divided into 3 groups (A, B and C). The patients in Group A (n = 4) were fitted with an intra-oral corrosion-resistant nickel-chromium Alloy A; the patients of Group B (n = 5) received a more corrosion prone nickel-chromium Alloy B and in Group C (n = 6) strongly corroding pure nickel was used. A corrosion-resistant foil of pure palladium was placed on the contralateral side. Reactivity of pure nickel foil was also tested on the skin in Group C. Immunohistological examination of the oral mucosa on the test and reference sides was performed with monoclonal antibodies directed against T-lymphocyte subsets and Langerhans cells (LC). The results showed that at the pure nickel site the LC did increase significantly in the connective tissue (approx. 4X) of the oral mucosa. However, statistical analysis of all 6 patients of Group C together showed no corresponding increase of LC in the epithelium at the site with the pure nickel, although a numerical increase of LC was noted in the epithelium adjacent to the pure nickel foil in 2 patients, which was remarkable. It can be concluded from statistical analysis that both the reference foils and the test foils can influence the number of suppressor/cytotoxic T-lymphocytes in the connective tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

The skin immune system (SIS): distribution and immunophenotype of lymphocyte subpopulations in normal human skin.

The complexity of immune response-associated cells present in normal human skin was recently redefined as the skin immune system (SIS). In the present study, the exact immunophenotypes of lymphocyte subpopulations with their localizations in normal human skin were determined quantitatively. B cells were not found to be present in normal human skin. Lymphocytes were always of T-cell type, and 90% of these T cells were clustered in 1-3 rows around postcapillary venules of the papillary vascular plexus or adjacent to cutaneous appendages. In such perivascular localizations, they were found to differ from their circulating counterparts in three ways. First, skin perivascular cells were found to be approximately evenly distributed over CD4+ inducer and CD8+ suppressor-cytotoxic T-cell subsets (mean CD4/CD8 ratio: papillary layer 0.96, reticular layer 0.99). Second, within the category of CD4+ inducer T cells, most were phenotyped as CD4+, 4B4+ helper inducer T lymphocytes, whereas CD4+, 2H4+ suppressor inducer T lymphocytes were found to be relatively rare (less than 5%). Third, the majority of skin perivascular T cells were activated as they expressed HLA-DR and interleukin 2 receptors. Intraepidermal, directly subepidermal, and other ("free") lymphocytes were mostly of the CD8+ suppressor-cytotoxic T-cell subset but accounted for less than 10% of the total number of lymphocytes. Intraepidermally localized T cells accounted for less than 2% of the total number of lymphocytes present in normal skin. Our results indicate that preferential perivascular localization of activated T lymphocytes is the characteristic of normal human skin. This might be a reflection of continuous antigen recognition upon endothelial cell presentation and/or continuous T cell-mediated endothelial cell activation thereby inducing enhanced antigen clearing by the skin's endothelium.

Different in situ distribution patterns of dendritic cells having Langerhans (T6+) and interdigitating (RFD1+) cell immunophenotype in psoriasis, atopic dermatitis, and other inflammatory dermatoses.

Dendritic cells bearing Langerhans cell (OKT6+) or interdigitating cell (RFD1+) immunophenotype may be regularly detected within the dermis of chronic skin diseases characterized by a lymphohistiocytic (lymphoreticular) infiltrate. These 2 subsets of antigen-presenting cells within the dermis of lesions of exacerbating chronic plaque psoriasis, exacerbating nummular dermatitis (discoid eczema), atopic dermatitis, allergic contact dermatitis, pityriasis rosea, lichen ruber planus, and cutaneous lupus erythematosus were quantified using computer-assisted morphometry. The mean dendrite length per dermal dendritic cell was significantly higher for RFD1 than for OKT6 (74.4 +/- 0.98 microns vs 70.0 +/- 1.26 microns: p = 0.0023). The mean dendrite length per dermal dendritic cell was remarkably constant for each marker in the various diagnostic categories studied. Disease-specific patterns of total dendrite length and number (expressed per 100 infiltrating mononuclear cells) of these 2 dendritic cell types within the subepidermal infiltrates were obtained. Pityriasis rosea was characterized by its unique high percentage of OKT6+ Langerhans cells. Atopic dermatitis and psoriasis had relatively high percentages of both RFD1+ interdigitating cells and OKT6+ Langerhans cells. Nummular dermatitis had an intermediate number and total dendrite length for OKT6, but was relatively low in RFD1+ cells. Allergic contact dermatitis, lichen planus, and lupus erythematosus had low numbers and dendrite lengths for both dendritic cell subsets. It is suggested that pityriasis rosea is characterized by an abnormal migration pattern of Langerhans cells. Psoriasis and atopic dermatitis may be examples of diseases in which skin-localized antigen-presenting and T-cell-inducing events are continuously taking place. The other diseases may reflect inflammatory processes in which local antigen presentation is less relevant to the tissue reaction.

Langerhans' cell population studies with OKT6 and HLA-DR monoclonal antibodies in vitiligo patients treated with oral phenylalanine loading and UVA irradiation.

In vitiligo patients, treated with oral phenylalanine loading combined with UVA irradiation (Phe-UVA), Langerhans' cells (LC) were counted in pigmented and depigmented skin. The LC, which were labelled with OKT6 and HLA-DR monoclonal antibodies, were expressed per linear mm epidermis. Before treatment the number of OKT6(+) cells was significantly increased in vitiliginous skin especially in the basal layer. Under treatment the number went down and was comparable to normal skin. When using HLA-DR labelling the number of LC increased in vitiliginous skin which had been treated with Phe-UVA. The influence of Phe-UVA on the shift of LC subpopulations is discussed.

Psoriasis infiltrating cell immunophenotype: changes induced by PUVA or corticosteroid treatment in T-cell subsets, Langerhans' cells and interdigitating cells.

The effects of PUVA or corticosteroid treatment on the distribution pattern of immunocompetent cells in psoriasis symptomatic skin were investigated. A total of 29 biopsies, taken before and a regular intervals during treatment, were studied in a two-stage immunoperoxidase technique using monoclonal antibodies directed against T cells, their major subsets, interdigitating cells, and Langerhans' cells. T cell exocytosis was not affected by PUVA or corticosteroid treatment. Instead, both treatment regimens led to an initial increase in interbasally localized T8+ suppressor/cytotoxic T cells. Increased T4/T8 ratios within the subepidermal infiltrates were restored to normal except in one case, who did not respond to PUVA-treatment. Epidermal and dermal T6+ Langerhans' cells decreased to almost absent. RFD1+ interdigitating cells had the same tendency, except in the PUVA-non-responding patient. Treatment of psoriasis with PUVA or corticosteroids thus results in a normalization of an initial immune imbalance in infiltrating immunocompetent cells. A working hypothesis on psoriasis immunopathogenesis and its restoration by treatment is presented.

Pityriasis rosea (Gibert): abnormal distribution pattern of antigen presenting cells in situ.

Pityriasis rosea is a skin disease which is obscure in its etiology and pathogenesis. We studied its immunopathology by immunophenotyping the inflammatory cells in situ using monoclonal antibodies that define leukocyte subsets. Findings as to T-cells and their major subsets did not reveal disease-specific data. Monocytes stained only rarely. Neither natural killer cells, B-cells nor plasma cells were ever found. An unexpected finding was the presence within the infiltrates and rarely within the epidermis of cells having the immunophenotype of interdigitating cells (RFD1+). Intense and dendritic staining with anti-T6 and anti-HLA-DR indicated Langerhans cells to be present in the dermal infiltrates, in between these infiltrates in the papillary dermis, and focally within the parakeratotic horny layer. This Langerhans' cell pattern provides evidence for dermal Langerhans cell compartmentalization and transepidermal Langerhans' cell elimination. Such a distribution indicates a change in Langerhans' cell migration processes in pityriasis rosea pathogenesis.

Acute disseminated histiocytosis-X: in situ immunophenotyping with monoclonal antibodies.

The proliferating skin cells in a case of acute disseminated histiocytosis-X (Abt-Letterer-Siwe disease) confirmed by electron microscopy, were characterized by a panel of monoclonal antibodies using an immunoperoxidase technique. The "histiocytes" were found to stain with OKT-6 (anti-T6) and anti-HLA-DR antibodies. Unexpectedly, slight staining was also observed with Leu 3a (anti-T4) and OKM-1. A proliferative process of T4, T6, HLA-DR, OKM-1 positive Langerhans' cells has not yet been described and may be specific for histiocytosis-X.

Immunocompetent cells in psoriasis. In situ immunophenotyping by monoclonal antibodies.

Immunocompetent cells in exacerbating untreated psoriasis vulgaris skin lesions were immunophenotypically studied by the application of a selection of monoclonal antibodies in a two-stage immunoperoxidase technique. Epidermal changes include: focal accumulation of immunoglobulins in the stratum corneum, as demonstrated by a mixture of monoclonal anti-kappa and anti-lambda antibodies; focal accumulation of OKM-1 positive but Mo-2 negative cells high in the epidermis, reflecting granulocytes in Munro's abscesses; a marked decrease in epidermal Langerhans cells with focal abnormal clumping and smaller dendrites, as demonstrated by monoclonal anti-HLA-DR and anti-T6 (OKT-6) antibodies; and, sporadic exocytosis of mainly T1 (Leu-1), T8 (Leu-2a) positive suppressor/cytotoxic T lymphocytes. The dermal infiltrates were found to consist mainly of partically activated T1 (Leu-1), T4 (Leu-3a) positive T-helper/inducer cells with a smaller compartment of T1 (Leu-1), T8 (Leu-2a) positive suppressor/cytotoxic lymphocytes. These cells were found in close apposition to T6 (OKT-6), HLA-DR positive Langerhans cells and further accompanied by a minor compartment of OKM-1, Mo-2 positive monocytes. No B-cells or plasma cells could be demonstrated in the dermis. Natural killer cells were observed only incidentally. These results fit best with the hypothesis that psoriasis is a chronic inflammatory condition as a result of persistent stimulation of T cells by immunogen(s) of epidermal origin.