Optimization of culture conditions for activation and large-scale expansion of human T lymphocytes for bispecific antibody-directed cellular immunotherapy

✍ Scribed by C. H. J. Lamers; R. J. van de Griend; E. Braakman; C. P. M. Ronteltap; J. Bénard; G. Stoter; J. W. Gratama; R. L. H. Bolhuis

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: French
• Weight: 775 KB
• Volume: 51
• Category: Article
• ISSN: 0020-7136
• DOI: 10.1002/ijc.2910510623

✦ Synopsis

We investigated the optimal culture conditions (i.e.. activation procedure, medium composition and type of culture vessel) for rapid in vitro expansion of large numbers (>5 X I 09) of blood T lymphocytes. These expanded lymphocytes can be targeted to be cytotoxic to ovarian carcinoma cells with a bispecific monoclonal antibody (BsAb) specific for CD3 and for the ovarian carcinoma-associated antigen MOv 18. Both phytohemagglutinin (PHA) and monoclonal antibody (MAb) CD3 induced rapid T-cell proliferation, although the growth kinetics after PHA activation were slightly faster. A 50-fold increase in cell number was obtained after I 4 and I6 days for PHA and CD3 MAb, respectively. The induction of BsAb-directed cytolysis was faster after CD3 MAb than after PHAactivation of lymphocytes, but became similar around day 20. A mixture of media consisting of 78% RPMl 1640, 20% AIM-V and 2% human plasma (Mix-med) yielded better results than 100% AIM-V medium. Culture of lymphocytes in polyolefin bags, compared with tissue culture flasks, or cryopreservation did not affect lymphocyte yield and function. In most cultures the proportion of CD8+ lymphocytes increased, suggesting a growth advantage of CD8+ over CD4+ lymphocytes in this culture system. A protocol employing PHA activation, Mix-med and polyolefin bags has been used successfully to activate and expand blood lymphocytes for the first 5 patients entered into a phase-1/11 clinical trial for the intraperitoneal treatment of ovarian carcinoma using CD3 X anti-MOv I8 BsAb-directed T lymphocytes.