Effects of trypsin on the in situ identification of epidermal cell membrane antigens

In this work the role of trypsin in revealing epidermal cell surface antigens, with the use of immunological markers, was investigated. Two monoclonal antibodies (MCA) were used, the first: D47, belongs to the first cluster of differentiation and recognizes a. membrane antigen of human thymocytes; the second HLA‐ABC‐m3, is an anti‐HLA‐B27 MCA. Preliminary treatment with various concentrations of trypsin was performed on frozen skin sections and followed by indirect immunofiuorescence. D47 reacted with epidermal dentritic cells only after trypsin pre‐treatment of skin sections. In addition a mild preliminary trypsinization was shown to increase in situ immunoreactivity of MCA HLA‐ABC‐m3 with epidermal cells. Best results were obtained when trypsin concentrations ranging from 2.5 to 5 μg/ml were applied for 10 min at 37°C. Preliminary trypsinizalion may be of interest for a better exposure of some surface antigens to immunohistochemical markers.

[1]  D. Schmitt,et al.  Reactivity of anti-HLA class I polymorphic monoclonal antibodies with normal human skin. , 1987, Hybridoma.

[2]  D. Schmitt,et al.  Subclustering of CD1 monoclonal antibodies based on the reactivity on human Langerhans cells. , 1986, Immunology letters.

[3]  M. Amiot,et al.  Heterogeneity of the first cluster of differentiation: characterization and epitopic mapping of three CD1 molecules on normal human thymus cells. , 1986, Journal of immunology.

[4]  Jean Kanitakis,et al.  Involucrin expression in adnexal skin tumours. An immunohistological study. , 1986, Virchows Archiv. A, Pathological anatomy and histopathology.

[5]  G. Pinkus,et al.  Optimal immunoreactivity of keratin proteins in formalin-fixed, paraffin-embedded tissue requires preliminary trypsinization. An immunoperoxidase study of various tumours using polyclonal and monoclonal antibodies. , 1985, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[6]  S. Gromkowski,et al.  Blocking of CTL-mediated killing by monoclonal antibodies to LFA-1 and Lyt-2, 3. II. Evidence that trypsin pretreatment of target cells removes a non-H-2 molecule important in killing. , 1985, Journal of immunology.

[7]  A. Bernard,et al.  The clusters of differentiation (CD) defined by the First International Workshop on Human Leucocyte Differentiation Antigens. , 1984, Human immunology.

[8]  M. Rijn,et al.  Human cutaneous dendritic cells express two glycoproteins T6 and M241 which are biochemically identical to those found on cortical thymocytes. , 1984, Human immunology.

[9]  E. Yonish-Rouach,et al.  Use of Tissue Distribution Studies to Determine the Specificity of Monoclonal Antilymphocyte Antibodies , 1984 .

[10]  A. Bernard,et al.  Diversity of Class I Molecules Heavy and Light Chains on Human Thymus Cells , 1984 .

[11]  S. Hakomori,et al.  Pattern of distribution of blood group antigens on human epidermal cells during maturation. , 1984, The Journal of investigative dermatology.

[12]  Jean Kanitakis,et al.  Anti‐D47: a monoclonal antibody reacting with the secretory cells of human eccrine sweat glands * , 1983, The British journal of dermatology.

[13]  J. Trapani,et al.  Description of a mouse monoclonal anti-HLA-B27 antibody HLA-ABC-m3. , 1983, Human immunology.

[14]  P. Wernet,et al.  A cytotoxic monoclonal IgM antibody (Tü 101) directed against an antigenic determinant shared between the HLA-A allospecificities A2 and A28. , 1983, Human immunology.

[15]  R. Fauchet,et al.  A monoclonal antibody to the HLA-A3 alloantigen. , 1982, Immunology letters.

[16]  S. Hsu,et al.  Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. , 1981, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[17]  C. Fenoglio,et al.  Reactivity of Langerhans cells with hybridoma antibody. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[18]  J. Fajumi.  Enzyme-induced modification of the surface properties of lymphoid cells in malignant disease. I. Effect of trypsin on rosette formation by lymphocytes in myelomatosis. , 1980, British Journal of Cancer.

[19]  R. Curran,et al.  Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections. , 1978, Journal of clinical pathology.

[20]  D. Fearon,et al.  Membrane sialic acid on target particles modulates their phagocytosis by a trypsin-sensitive mechanism on human monocytes. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Brozman.  Immunohistochemical analysis of formaldehyde- and trypsin- or pepsin-treated material. , 1978, Acta histochemica.