|Budget Amount *help
¥5,600,000 (Direct Cost : ¥5,600,000)
Fiscal Year 1995 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1994 : ¥4,900,000 (Direct Cost : ¥4,900,000)
We have developed a serum-free medium designated RDSF for the generation of LAK cells based on RD6F medium, which was originally developed as a serum-free medium for the growth of myeloma and hybridoma cells. The cytotoxic activity of LAK cells generated in RDSF against Raji, K562 and oral cancer cells, is 3-4 times that of LAK cells generated in medium containing 10% human type-AB serum. RDSF medium consisted of nutrient mixture supplemented with transferrin, 2-aminoethanol, 2-mercaptoethanol, sodium selenite and interleukin-2. In this study, we have found that insulin, which has been found to be the most important polypeptide hormone in surum-free media for animal cells, inhibited the generation of cytotoxic activity of LAK cells cultured from pheripheral blood lymphocytes. In addition, we have found that transferrin was an essential component for the growth and generation of LAK cells in serum-free culture. Thses results suggest that RDSF will be usefel for adoptive immunotherapy of cancer and studying factors involved in the growth and differentiatiation of LAK cells.
In addition, we have studied effect of monoclonal antibody (MoAb) to EGF-r designated as 12-93 on the growth of squamous cell carcinoma cells (SCC) and salivary gland adenocarcinoma cells (SAC) in vivo, and targeting immunotherapy using 12-93 MoAb-conjugated LAK cells against oral cancer cells in vitro, A 12-93 MoAb inhibited the growth og SCC and SAC in vivo. The 12-93 conjugated LAK cells showed significantly enhanced cytolysis of 12-93 reactive A431 and HSG cells but not Raji cells which dose not react with 12-93. These results indicated that 12-93 will be aviable altemative to tumor specific MoAb and 12-93-conjugated LAK cells may provide an effective strategy for targeting adoptive immunotherapy of cancer.