Functional analysis of Ia antigen-bearing keratinocytes: mixed skin lymphocyte culture between Ia antigen-bearing Pam 212 cells and allogeneic and syngeneic splenic T cells.
Keratinocytes express Ia antigens in various skin disorders, although the biological role of these Ia antigen-bearing (Ia+) keratinocytes remains unclear. We induced Ia antigens on Pam 212 murine keratinocyte cell line by interferon-gamma(IFN-gamma) and using these cells, we performed the mixed skin lymphocyte culture with syngeneic BALB/c or allogeneic C3H/He splenic T cells. Unexpectedly, Pam 212 cells were found to stimulate both syngeneic and allogeneic T cells irrespective of IFN-gamma treatment. However, both syngeneic and allogeneic T cells cultured with IFN-gamma-treated Pam 212 cells incorporated [3H]thymidine much more actively than those cultured with IFN-gamma-untreated Pam 212 cells. This stimulation was not inhibited by monoclonal anti-I-Ad antibody. Analysis of the responding T cells demonstrated that the syngeneic T-cell stimulation by IFN-gamma-treated Pam 212 cells occurred in both purified Lyt 1-T cells and Lyt 2- T cells. Furthermore, we found that the T cells cultured with the IFN-gamma-treated cells were composed of two morphologically different types of cells. Determination of their surface phenotype showed that the small cell population consisted of 57% Thy-1+, 23% Lyt-1+, 6% Lyt-2+, and 9% asialo-GM1+ cells, while the large cells consisted of 53% Thy-1+, 15% Lyt-1+, 9% Lyt-2+, and 24% asialo-GM1+ cells. These findings suggest that IFN-gamma-treated Pam 212 cells could stimulate more than one kind of splenic T cell populations.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002478:Cells, Cultured; D004261:DNA Replication; D005260:Female; D000949:Histocompatibility Antigens Class II / Q000032:analysis*; D007371:Interferon-gamma / Q000494:pharmacology; D007518:Isoantibodies / Q000276:immunology; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D012867:Skin / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D013154:Spleen / Q000166:cytology*; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000949:Histocompatibility Antigens Class II; D007518:Isoantibodies; D007371:Interferon-gamma