<Original Report>A NEW METHOD FOR CYTOTOXICITY IN VITRO TEST OF CELLULAR IMMUNITY

Abstract

A new quantitative in vitro test for the direct measurement of cytotoxicity on confluent and complete monolayer of SMT cells was designed with Micro Test Plate. The tumor used develops with high frequency in the DBA/2Ha-DD mouse, and is specific for this mouse since it grows in syngeneic host but not in DBA/2J or C57B1/6Ja mice. 1) With 10% serum supplement regardless of whether fetal calf serum or syngeneic mouse serum, more than 60% of the lymphocytes were viable at 48 hours culture after plating. 2) Number of blastic transformation of lymphocytes was increasing with the percentage of the fetal calf serum in 3×E medium. 3) The activity of immune lymphocytes against SMT monolayer was cell number-dependant which was intraperitoneally immunized. 4) The cytotoxicity of immune lymphocytes was greatest with cells from C57B1/6Ja mice, less marked with cells from DBA/2J mice and least with cells from DBA/2Ha-DD mice. Moreover, its cytotoxicity was greater against SMT monolayer than against MKC monolayer. 5) Therefore, it can be concluded that cellular immunity can be demonstrated between against allo-antigen and against tumor specific transplantation antigen of SMT. This method can, also, be consider to be a good one for cytotoxicity in vitro test of cellular immunity.