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

1987; Wiley; Volume: 14; Issue: 6 Linguagem: Inglês

10.1111/j.1600-0560.1987.tb01533.x

1600-0560

Giovanna Zambruno, V. Gielen, Daniel Schmitt, Colette Dezutter‐Dambuyant, J Thivolet,

Immunotherapy and Immune Responses

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.