Abstract
The cytolysis of 3H-proline-labelled tumour cells growing in monolayer by syngeneic immune lymphocytes has been studied in the murine sarcoma virus (MSV) system. Results show that the proline assay (PA) is a convenient way to reveal the activity of cytolytic T lymphocytes against FMR-like antigens. Using the same effector and target cells, the classical chromium-release test (CRT) fails to reveal any cytolytic activity, and the visual microcytotoxicity assay as well as several derived isotopic methods are known to reveal mainly non-specific reactions due to non-T effector cells. The PA, therefore, appears to be a useful method for testing an antitumour reaction against tumour cells in monolayer. The results are, however, different from those obtained in the CRT using the same effector cells but lymphoma cells in suspension as targets, the major discrepancies being the following: (a) the PA does not provide truly quantitative data, due to the very high lymphoid: effector cell ratios needed in this test; (b) unexpected patterns of antigenic specificities are sometimes detected in PA; (c) a non-specific natural killer activity of non-T cells is frequently detected in the PA, masking at low lymphoid: target cell ratios the T-dependent specific cytolysis; (d) the H-2 restriction of the cytolytic T-cell activity is poorly detected in PA, whereas the role of H-2 antigens is clearly shown by blocking experiments using anti-H-2 antibodies.
Highlights
Summary.-The cytolysis of 3H-proline-labelled tumour cells growing in monolayer by syngeneic immune lymphocytes has been studied in the murine sarcoma virus (MSV) system
ONE of the major problems facing tumour immunologists studying cellmediated reactions results from the disparity in the results with different in vitro cytotoxicity assays. Such disparity could be caused by variations in methodological parameters, which include the nature of the target cells and the length of the incubation period in vitro with the effector cells
The cytostatic phenomenon mediated by non-T cells being, at least for the main part, non-specific (Owen & Seeger, 1973; Senik et al, 1974) it is often difficult to assess the specificity of the reactions measured by such methods
Summary
Summary.-The cytolysis of 3H-proline-labelled tumour cells growing in monolayer by syngeneic immune lymphocytes has been studied in the murine sarcoma virus (MSV) system. ONE of the major problems facing tumour immunologists studying cellmediated reactions results from the disparity in the results with different in vitro cytotoxicity assays Such disparity could be caused by variations in methodological parameters, which include the nature of the target cells and the length of the incubation period in vitro with the effector cells. More antigen-specific reactions are generally obtained using short-term assays with ascitic lymphoma cells or tumour cells cultivated in suspension as targets Under these conditions, very clear results can be obtained in the MSV system, regardless of the isotopic marker e.g., 51Cr (Leclerc et al, 1972), 125IUdR (Oldham & Herberman, 1973) or 3H-proline (Shiku et al, 1975; Oldham et al, 1977). They are, much less antitumour cell-mediated reactions in efficient with target cells in monolayers. different assays
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