Lectin-dependent cell-mediated cytotoxicity. A new and simple method to quantitate cytotoxic T cell activity in dogs.
Alloimmune T lymphocytes kill a wide spectrum of target cells, even in the absence of alloantigen recognition, providing that a lectin is incorporated into the assay medium. In these studies we have modified the lectin-dependent cell-mediated cytotoxicity (LDCMC) assay to evaluate the cytotoxic activity of canine T cells using a human B lymphoblastoid cell line (Raji) as the target. Canine peripheral blood mononuclear cells activated in vitro in mixed leukocyte cultures or by concanavalin A (Con A) killed 51Cr-labeled Raji target cells (up to 70% in 4 hr) when Con A was included in the assay. T cells expressed cytotoxic activity only when activated and when con A was present in the assay. Canine T cells activated in vivo also exerted cytotoxic activity in the LDCMC assay. T cells obtained from a rejecting allografted lung by bronchoalveolar lavage displayed considerably higher cytotoxic activity than did T cells obtained from the contralateral normal lung or peripheral blood of the same dog. In contrast, T cells from the lungs and blood of normal dogs and lung-allografted dogs without rejection exerted no cytotoxic activity. We conclude that the LDCMC assay using xenogeneic Raji cells as targets is a valid measure of the cytotoxic activity of canine T cells. Its major advantages include the fact that it is simple to perform and can be used when specific target cells are not available.
D000818:Animals; D002469:Cell Separation; D003208:Concanavalin A / Q000494:pharmacology*; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D004285:Dogs; D004306:Dose-Response Relationship, Immunologic; D007111:Immunity, Cellular; D008168:Lung / Q000166:cytology / Q000276:immunology; D016040:Lung Transplantation; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D013601:T-Lymphocytes / Q000276:immunology*
D003208:Concanavalin A