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Biomedical subjects

A Scheynius

Publications and source records attributed to A Scheynius.

At least 91 records · Page 5Linked to original sources

T-cell response to purified protein derivative after removal of Langerhans' cells from epidermal cell suspensions containing keratinocytes expressing class II transplantation antigens.

In a previous study we observed that human epidermal cell (EC) suspensions containing HLA-DR-expressing keratinocytes showed an amplified T-cell response to purified protein derivative (PPD). To evaluate further the possible immunological importance of class II transplantation antigens on keratinocytes we have compared the T-cell response to PPD in the presence of the following stimulator cells: EC suspensions from normal skin, or EC from tuberculin-reactive skin with or without removal of Langerhans' cells. The proliferation of purified T lymphocytes from peripheral blood in response to PPD in the presence of various concentrations of autologous EC was measured by [3H]thymidine incorporation on day 6. In 3 experiments out of 4 the EC from tuberculin-reactive skin, containing 28-76% HLA-DR-expressing cells as judged by immunocytochemistry (which also revealed fairly numerous HLA-DQ/-DP-expressing keratinocytes and a slight increase in CD36- and CD4- but not CD1-expressing cells), induced a more pronounced T-cell response to PPD than did normal EC. This was not the case in the fourth experiment, in which a small number of HLA-DR-(15%) and few if any HLA-DQ-/-DP-expressing keratinocytes were found. Immunomagnetic removal of CD1-reactive Langerhans' cells from the tuberculin-reactive EC suspensions resulted in a reduction of the T-cell response to PPD, in most cases down to background level (T cells alone + PPD). This study does not support the hypothesis that HLA-DR-expressing keratinocytes can in themselves act as antigen-presenting cells.

Adult

Low dosage alpha-interferon treatment in patients with advanced cutaneous T-cell lymphoma.

Treatment with high dose (10-50 x 10(6) IU/m2 three times a week) of alpha-interferon (alpha-IFN) has been shown to induce remissions in about 50% of patients with cutaneous T-cell lymphoma. The optimal alpha-IFN dose, however, is not known. 4 patients with advanced cutaneous T-cell lymphoma were therefore treated with alpha-IFN in low doses of 2-4 x 10(6) IU/m2 three times a wk. 1 complete, 1 partial and 1 minor remission were observed. Skin biopsies before and during treatment were taken from 2 of the patients and showed improvement. Low doses of alpha-IFN can thus induce remission in patients with cutaneous T-cell lymphoma.

Adult

Dendritic cells and macrophages expressing class II antigens in the normal rat incisor pulp.

This study has identified and characterized class II (Ia) antigen-expressing cells in the normal rat incisor pulp by immunohistochemistry and flow cytometry. Two types of Ia-expressing cells occurred: one with a pronounced dendritic appearance located primarily in the periphery of the pulp, and one with morphological characteristics similar to those of macrophages. The latter cells were mainly observed in the central portion of the pulp. A numerical ratio of 1:4 was established between the two cell types. The existence of Ia-expressing cells suggests an inherent capacity of the pulp to process and present foreign antigens.

Animals

Enhancement of DTH reaction and inhibition of the expression of class II transplantation antigens by in vivo treatment with antibodies against gamma-interferon.

During the delayed type of hypersensitivity (DTH) reaction in the skin, class II transplantation antigens are expressed on the keratinocytes. This induction is attributed to the action of gamma-interferon (IFN-gamma). We have now studied the influence of antibodies against IFN-gamma on the DTH-reaction. Lewis rats were sensitized to 2,4-dinitro-1-fluorobenzene (DNFB) and challenged 5 days later with DNFB on the ears. Immediately before challenge each animal in one group (n = 16) was given 1 mg of mouse monoclonal antibodies (MoAb) against rats IFN-gamma, denoted DB-1, intraperitoneally (i.p.), another group (n = 15), 1 mg of an irrelevant MoAb and a third group (n = 11) was left untreated. The ear thickness was measured with a micrometer, before challenge and after 24, 48 and 72 h. At 72 h all rats were killed, the ears cut off, snap frozen and stained with immunoperoxidase using MoAbs OX 6 and OX 17, directed against rat class II antigens. The DB-1 treated group was found to have a larger ear swelling that was statistically significant at each time point compared with the other two groups. Furthermore, the animals given DB-1 showed class II antigen expression on Langerhans' cells, but almost none on keratinocytes. In contrast, the rats in the two other groups displayed a moderate to strong expression of class II antigens on keratinocytes as well as on Langerhans' cells. It is concluded that DB-1 can inhibit class II antigen expression on keratinocytes during the DTH- reaction and also enhance the local response.

Animals

Removal of Langerhans cells from human epidermal cell suspensions by immunomagnetic particles.

The possibility of eliminating the class II antigen-expressing and antigen-presenting Langerhans cells from normal human epidermal cell suspensions was investigated. Cell suspensions containing 1.7-2.8% Langerhans cells were prepared by enzyme treatment of human skin obtained at plastic surgery. The cells were incubated with mouse monoclonal antibodies directed against the CD1 antigen present on Langerhans cells and were further incubated with magnetic particles, 4.5 micron in diameter, coated with sheep anti-mouse IgG. The optimal ratio between particles and target cells was found to be 40:1. The rosetted Langerhans cells were removed by a cobalt-samarium magnet. In six experiments immunoperoxidase staining revealed 0-0.1% (mean 0.03%) CD1-reactive Langerhans cells in the depleted cell fraction. The alloantigen presenting capacity of the depleted cell fraction, measured by [3H]thymidine incorporation, was abolished.

Antigen-Presenting Cells

A simple in vitro technique for studies on induction of class II transplantation antigens on keratinocytes.

A new technique is described for evaluation of the induction and kinetics of class II transplantation antigens on keratinocytes. By culturing small rat or human skin specimens for 2-5 days in media containing gamma-interferon, class II antigens were detected on keratinocytes by immunohistochemistry. This technique is rapid and technically simple compared to keratinocyte cultures and raises the possibility of studying other cells in the skin.

Animals

Amplification of T-cell response to PPD by epidermal cell suspensions containing HLA-DR-expressing keratinocytes.

The biological importance of the presence of class II transplantation antigens on highly differentiated epithelial cells such as keratinocytes in certain conditions, is still unknown. We have therefore investigated the antigen-presenting capacity of separated human epidermal cells obtained from tuberculin-reactive skin 6 days after intradermal injection of purified protein derivative (PPD). Earlier studies have shown a high percentage of HLA-DR-expressing keratinocytes at this time. Peripheral adherent blood cells were used as control stimulator cells and highly purified peripheral blood T lymphocytes as responder cells. The T-cell proliferation in response to PPD in the presence of autologous epidermal cells from normal and tuberculin-reactive skin was measured by [3H]thymidine incorporation on day 6. The latter cell population, 76-86% of which consisted of HLA-DR-expressing cells as judged by immunocytochemistry, induced a greater T-cell response to PPD than do normal epidermal cells. This discrepancy in the T-cell proliferation could not be explained by a difference in the numbers of anti-Leu 6 or anti-HLA-DQ-reactive Langerhans cells. The present data indicate that epidermal cell suspensions containing HLA-DR-expressing keratinocytes induce a greater T-cell response to PPD than do normal epidermal cells.

Adult

Effects of monoclonal anti-T cell antibodies on rat cardiac allografts.

Monoclonal antibodies reactive with different T lymphocyte antigens were administered to rats receiving heart allografts. Ox 19 antibodies (directed to the rat Ly 1 equivalent) and Ox 8 antibodies (directed to the rat CD8 equivalent) both prolonged graft survival, whereas W3/25 (anti-CD4), Ox 6 (anti-Ia), and W3/13 (anti-pan T) antibodies did not affect graft rejection. Immunohistological studies were carried out on spleen and graft specimens in order to analyse further the mechanisms behind the prolongation of graft survival. The observed almost complete absence of Ox 8-reactive cells in the spleen after treatment with Ox 8 antibodies corroborates earlier observations that injection of moderate amounts of Ox 8 antibodies leads to complete elimination of suppressor/cytotoxic T cells from peripheral lymphoid organs and blood. The present data on graft survival therefore both support the notion that suppressor/cytotoxic T cells are involved in graft rejection, and suggest that these cells are not the only ones involved. An unexpected and as yet unexplained finding was that Ox 8-reactive molecules were found in large numbers on various inflammatory cells as well as on certain myocytes in the grafted hearts that had experienced a prolonged graft survival due to treatment with Ox 8 or Ox 19 antibodies.

Animals

In vitro testing of contact sensitivity.

A new in vitro technique for the testing for contact sensitivity is described. The method is based upon the fact that in response to immunological stimuli, keratinocytes can start to synthesize and express class II histocompatibility antigens. Nickel sulphate (NiSO4) stimulated lymphocytes from nickel-sensitive persons, and the supernatant of these cells thus induced expression of HLA-DR on keratinocytes when co-cultured with autologous normal skin biopsies. This was in contrast to what was the case in healthy controls.

Dermatitis, Contact

Enzyme-linked immunosorbent assay of H+,K+-ATPase, the parietal cell antigen.

Vesicular membranes, purified from porcine gastric mucosa and rich in H+,K+-ATPase, were used to establish an enzyme-linked immunosorbent assay (ELISA) for determinations of parietal cell autoantibodies. Results obtained with the ELISA correlated well with standard immunofluorescence determinations of parietal cell antibodies based on frozen sections of rat stomach. The ELISA however was about 10-fold more sensitive than the immunofluorescence method and had high specificity. Intra- and interassay coefficients of variation, determined with a patient sera of average positivity, were 5.5% and 18%, respectively. The ELISA detected antibody binding in 23 out of 26 sera from patients with known autoimmune atrophic gastritis, in five of 25 sera with autoimmune thyroiditis, in five of 20 sera from patients with Graves' disease, in three out of 20 sera from patients with atoxic nodular goitre, in six of 20 sera of patients with primary biliary cirrhosis, in two out of 20 sera of patients with active duodenal ulcer, in two out of 20 sera with detectable antinuclear antibodies, and in one out of 20 sera with detectable rheumatoid factor. Data determined by an ELISA based on a gastric vesicular membrane preparation of human origin correlated well (r = 0.79, P less than 0.001) to those obtained by the standard ELISA based on porcine membrane material. The assay should be well suited for routine determinations of parietal cell antibodies in investigations of autoimmune gastritis and multiple organ autoimmune endocrinopathies.

Adenosine Triphosphatases

An immunologic and cultural study of Pityrosporum folliculitis.

In patients with Pityrosporum folliculitis the mean serum antibody titer against Pityrosporum orbiculare was significantly higher than in healthy control subjects (p less than 0.01). The mean number of P. orbiculare organisms per square centimeter cultured from normal-looking skin in patients was not significantly higher than the number cultured from normal-looking skin in control subjects. Results of prick tests against P. orbiculare extract were negative or weak, indicating that patients with Pityrosporum folliculitis had no type I hypersensitivity against P. orbiculare. Immunohistochemical staining of skin lesions showed perivascular dermal cell infiltrates near the hair follicles dominated by anti-Leu 3a-reactive T lymphocytes. Human lymphocyte antigens with the DR locus, but not those with the DQ locus, on keratinocytes were observed in one case.

Adult

In vivo induction of Ia antigens on rat keratinocytes by gamma-interferon.

Recombinant rat IFN-gamma was found to induce Ia antigen expression on rat keratinocytes in vivo. Strong expression of Ia antigen on keratinocytes of rat ears was produced after intradermal injections of 10,000 U IFN-gamma once daily on 3 successive days. The expression was still pronounced 2 days after the last injection, but had disappeared 4 days later.

Animals

Expression of HLA-DQ antigens on keratinocytes in Borrelia spirochete-induced skin lesions.

Skin biopsies were investigated with two different immunohistochemical techniques, thus revealing HLA-DQ antigens on HLA-DR-expressing keratinocytes in the late skin manifestations of a Borrelia spirochete infection. In the early skin lesions only HLA-DR antigens were present on the keratinocytes. The invariant gamma chain of class II transplantation antigens was observed on keratinocytes in 1:5 of the late cases. Upon penicillin treatment detectable HLA-DR and HLA-DQ antigens disappeared completely from the keratinocytes. Furthermore, the mononuclear cell infiltrates dominated by anti-Leu 1 and anti-Leu 3a-reactive cells and containing many cells with markers for activation (HLA-DR, HLA-DQ, transferrin, and interleukin 2 receptors) diminished markedly. The possibility that the expression of different class II transplantation antigens on keratinocytes might reflect separate functional demands of these cells or an altered immunological reactivity in the host, is discussed. The precise functional role of the temporary expression of the class II antigens on non-lymphoid cells, however, remains an enigma.

Acrodermatitis

Phenotypic difference between allergic and irritant patch test reactions in man.

Cellular responses in allergic and irritant contact dermatitis were analysed in situ using an immunohistochemical double staining technique with the aim of uncovering phenotypical differences of diagnostic importance. Allergic and irritant patch test reactions were elicited in 9 individuals using the Finn chamber technique. Thirty-nine skin biopsies from these reactions and from petrolatum controls were obtained 4 to 20 days after the test applications. Cell infiltrates were present throughout the observation period in both allergic and irritant reactions, but were usually greater in the former. In both types of reaction, anti-Leu 3a reactive cells predominated over anti-Leu 2a reactive cells. HLA-DR expression on keratinocytes was found in 9 of 14 allergic reactions, but not in irritant reactions or control areas. HLA-DQ antigens were not detected on keratinocytes. The presence of HLA-DR antigens on keratinocytes may reflect an immunological response of the allergic reactions, and thus be of diagnostic relevance.

Adult

Expression of Class II transplantation antigens by epithelial cells in oral candidosis, oral lichen planus and gingivitis.

Biopsies from normal oral mucosa and oral mucosa affected by candidosis, lichen planus or gingivitis were compared with respect to the expression of two Class II transplantation antigens, HLA-DR and HLA-DQ, by epithelial cells and the relationship of these antigens to the distribution and frequency of T-lymphocytes. Indirect immunohistochemistry with different mouse monoclonal antibodies was used on frozen and acetone-fixed sections. To evaluate the results, a score system based upon the expression of the Class II transplantation antigens by epithelial cells and the frequency of T-lymphocytes was used. In oral candidosis there was a marked expression of HLA-DR antigens throughout the epithelium. In addition, this type of epithelium was the only one that expressed HLA-DQ antigens. An intense intraepithelial infiltration of T-lymphocytes was observed. Oral lichen planus and gingivitis did, to a much lesser extent, cause the expression HLA-DR antigens by the epithelial cells. In both lesions, the number of T-lymphocytes within the epithelium did not exceed the number found in epithelium of normal mucosa. In these types of lesions, the subepithelial infiltrate varied in intensity but was mainly composed of T-lymphocytes reactive with anti-Leu 3a antibodies. The results of the present study imply that epithelial expression of the two different Class II antigens are related to the frequency of the T-lymphocytes and to the proximity of these cells to the epithelial cells.

Adult