Development of Culture system for LAK cells and its application to adoptive immunotheraphy for lung cancer.

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For application with surgical adjuvant therapy of cancer. we have designed a unique regimen of adopuve immunotherapy with lymphokineacuvated killer (LAK) cells and recombinant interleukin-2 (IL-2). The regimen features the prolonged (6 consecutive days) subcutaneous administration of a low-dose IL-2 and the transfer of ex vivo generated LAK cells from regional lymph node lymphocytes, obtained at the time of operations. According to this regimen. 19 patients with pathological stage-1 lung cancer received immunotheraph about 2 weeks after pulmonary lobectomy. Clinical toxicities included fever. fatigue. <5% weight gain. and edema.All toxicities reversed within 4 days after the complection of therapy. In 13 patients. peripheral blood lymphecytes obtained 24h after therapy exhibited in vitro LAK activity. Thus the regimen is considered to be well-tolerable and immunologically active, in regard to the postoperative state of the patients.To improve the routine performance of the generation and expansion of LAK cells. we have developed a new culture system (LIFROC Liquid-Filled Rotary Culture : formaly designated JCC device). The LIFROC system is essentially based on dialysis culture. A dialysis membrane divides the culture vessel from the outside circulation of basal medium. The culture vessel is rotated for the proper dispersion of cells and supplied with oxygen through a silicon tube.With this LIFROC system. the cell density reached a maximum 2.7x10^7 cells/ml with greater than 90% viability and a substantial level of LAK activity. In comparison with a convenstonal batchwise culture system suing tissue culture dishes. the LIFROC system was able to reduce the consumption of basal medium. IL-2. and serum by 20%, 84%, and96%, respectively.Culture vessel, reservoir, and tools for the recovery of LAK cells from culture medium were devised to be disposable. This culture system is apphed with safe to culture LAK cells for adoptive immunotheraphy.