Solid‐phase Anti‐CD3 Antibody Activation and Cryopreservation of Human Tumor‐infiltrating Lymphocytes Derived from Epithelial Ovarian Cancer

The effect of solid‐phase anti‐CD3 antibody activation and cryopreservation was evaluated on thirteen samples of tumor‐infiltrating lymphocytes (TILs) derived from epithelial ovarian cancer. Seven preparations of TILs were cultured with or without solid‐phase anti‐CD3 antibody in addition to 100 units/ml of recombinant interleukin‐2 (rIL‐2). The proliferation rate of all of the seven TIL preparations stimulated by anti‐CD3 antibody on the fourth or fifth day of culture was 3.4 to 9.8 times greater than that of lymphocytes cultured with rIL‐2 alone. Furthermore, in an experiment with five TIL samples activated with anti‐CD3 antibody, three of them showed augmented cytotoxic activity against autologous fresh tumor cells. The population of CD3(+)/CD8(+) TILs was increased after 4–5 weeks of cultivation and CD8(+) lymphocytes amounted to over 70% in all of seven preparations tested, whereas two of seven preparations not activated by anti‐CD3 antibody were CD3(+)/CD4(+)‐dominant. In addition, nine preparations of TILs cultured with rIL‐2 were cryopreserved for several weeks; after recovery from cryopreservation, no major change was observed in cell surface markers, in growth rate or in cytotoxic activity. These results suggest that cryopreserved and/or anti‐CD3 antibody‐activated lymphocytes could conveniently be employed in a clinical trial of adoptive immunotherapy employing TIL.