[HTML][HTML] 51 Chromium-release assay for cell-mediated cytotoxicity of human leukemia and lymphoid tissue-culture cells

JL McCoy, RB Herberman, EB Rosenberg… - Natl Cancer Inst …, 1973 - books.google.com
JL McCoy, RB Herberman, EB Rosenberg, FC Donnelly, PH Levine, C Alford
Natl Cancer Inst Monogr, 1973books.google.com
Cellular immune reactions to antigens associated with human leukemia were studied by a
lymphocyte cytotoxicity assay employing the release of 51chromium (51Cr) from labeled
cells of leukemic patients. Low-grade, but significantly positive, cellular cytotoxicity was
obtained by the reaction of autologous or allogeneic lymphocytes from patients and from
normal adult donors against cells from leukemic patients. Similar reactivity was not found
against target cells from normal donors. The results suggest that human leukemia cells …
Summary
Cellular immune reactions to antigens associated with human leukemia were studied by a lymphocyte cytotoxicity assay employing the release of 51chromium (51Cr) from labeled cells of leukemic patients. Low-grade, but significantly positive, cellular cytotoxicity was obtained by the reaction of autologous or allogeneic lymphocytes from patients and from normal adult donors against cells from leukemic patients. Similar reactivity was not found against target cells from normal donors. The results suggest that human leukemia cells possess tumor-associated antigens. Studies to establish immunologic specificity of the positive reactions were technically difficult because of the low reactivity seen and because of difficult logistic problems in repeating the tests. Therefore, to study a variety of parameters that might help establish the specificity of the assay in human systems, human lymphoid cell lines derived from patients with tumors and from normal individuals were used as targets. Many of these (including F-265 and NC37) derived from normal lymphocytes were highly susceptible to lysis by lymphocytes of most normal donors. Agents, such as X-irradiation, actinomycin D, 3'5'cyclic-AMP, and concanavalin A, blocked either completely or partially the cytolytic activity of lymphocytes against F-265 cells. Target cell inhibition tests revealed that addition of graded doses of unlabeled F-265 cells always inhibited 51Cr release from labeled F-265 cells. Similarly, inhibition of cytotoxicity of F-265 was also inhibited by some other cultured lymphoid cells including NC37, but was not inhibited by at least one other cultured lymphoid cell line (Stuck). Normal human lymphocytes stimulated in vitro by phytohemagglutinin were neither effective as inhibitor cells in the F-265 system nor were they directly lysed by lymphocytes of normal individuals. Finally, F-265 cells grown in media containing human sera instead of fetal bovine serum were effective targets in the assay. The results suggest that true cell-mediated immunologic reactions cause the lysis of human tissue-culture lymphoid cells by normal donor lymphocytes. The relevant antigen (s) appears neither to be" blast" associated nor to be related to antigens of fetal bovine serum. Hopefully, this system will be a useful model for defining the specificity of the cellular reactivity against leukemic cells.-Natl Cancer Inst Monogr 37: 59-67, 1973.
books.google.com