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KMID : 0360319890210010035
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Journal of Korean Cancer Research Association
1989 Volume.21 No. 1 p.35 ~ p.50
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In Vitro Activation and Expansion of Human Tumor-Infiltrating Lymphocytes(TIL) with Intreleukin 2(IL2) and Their Antitumor-Cytotoxic Properties Against Autologous Tumor Cells
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최규철/Choeh, Kyuchul
남상윤/박재경/하윤문/최용묵/안창열/Nam, Sang Yun/Park, Jai Kyung/Ha, Youn Mun/Choi, Yong-Mook/Ahn, Chang Il
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Abstract
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To determine the effect of interleukin 2 (IL 2) on human tumor-infiltrating lymphocytes (TIL) from surgically excised solid tumors and to characterize the antitumor-cytotoxicity of activated TIL comparing with lymphokine-activated killer (LAK) cells generated from peripheral blood mononuclear cells, this study was performed.
Single cell suspensions of TIL and tumor cells were prepared from 20 surgical specimens (12 stomach cancers, 4 breast cancers, 1 lung cancer, 1 colon cancer, 1 ovarian cancer, and 1 cervix cancer) by mechanical and enzymatic means. And freshly isolated TIL were separated from tumor cells by fishing method.
The amount of TIL obtained by the above means was 7.1 x 10^(5)±6.7 (SD) cells/㎤ of tumor mass with range of 0.8~27.5 x 10^(5), The TIL prepatrations contaminated partially with tumor cells (10~20%) were cultured in IL 2 containing media. Cell numbers were reduced from the initial seeding density until about 7 days of culture. After 10 days of culture with IL 2, TIL increased in number with a concomitant disappearance of tumor cells, whereas there were slow decreases of lymphocytes and increases of tumor cells in control cultures.
TIL were expanded between 0.5 and 118,000 fold increases for 10 to 34 days (2~9 subcultures). Partially purified natural IL 2-induced expansions of TIL were more rapid than highly purified recombinant IL 2-induced expansions, and showed 10 times high for 18 days of culture. The majority of TIL were T lymphocytes with range of 56~68.8% of Leu4+, 25.0~40, 8%of Leu2a+, and 0~51. 5% of Leu3a+cells. With continued in vitro expansion for 14~20 days, there was a concomitant increase in T lymphocytes (Leu4+) to 71~76.8 % but the changes of phenotypes of Leu2a+ and Leu3a+ were variable.
TIL were tested for cytotoxicity against fresh autologous and allogenic tumor cells, and Raji and K562 targets in a 4-hr ^(51)Cr release assay. IL 2-activated TIL showed more potent cytotoxicity against autologous tumor targets than allogenic (P<0.05).
And cytotoxic activity of activated TIL in killing autologous tumor targets was higher level than that of LAK cells, but the defference was not statistically significant (0.05In conclusion, TIL were induced in proliferation and augmented in their cytotoxixity by IL 2. IL 2-activated TIL preparations showed an apparent specific cytotoxicity and exhibited the combined preperties of cytotoxic T lymphocytes and LAK cells.
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