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J Korean Cancer Assoc > Volume 21(2); 1989 > Article
Journal of the Korean Cancer Association 1989;21(2): 339-354.
림포킨활성살해세포의 유도기와 세포독성기에서의 혈청의 영향
권오중, 김진복
Serum Effect on the Indurtive Phase and Cytotoxic Phase of Lymphockine - activated Killer Cell
Oh Hoong Kwon, Jin Pok Kim
LAK (lymphokine activated killer) cells cytotoxic for human colon carcim>ma cell line SUN (:. i were generated in vitro when recombinant interleukin-2 (IL-2) was incubated with peripheral biood mononuclear cells from normal person and stomach cancer patient. This study was designed to establish the optimal condition for LAK activation and to evaluate the effect of fresh human serum on the LAK activity. Peripheral blood mononuclear cells were incubated in RPMI-1640 with 10ro FCS (fetal calf serum) of 10% AHS (autologous human serum) at the concentration of 1 unit/ml of recombinant IL-2. Cytotoxicity against SNU-C5 was measured by tetrazolium-based colorimetric (MTT) assay. The cytotoxic response first appeared significantly after 3 days of culture with recombinatn JL-2, and it peaked after 5 days and declined after 7 days of incubation. During 5 dyas culture, recombinant II.-2 induced the potent LAK activity at the concentration of 1 unit/ml. After 4 days incubation. almost all target cells (SNU-C5) were killed bv the LAK of 5 davs culture at the effector: target cell ratio of 10:l. The level of LAK activity generated was significantly reduced (P<0.00l) when 10% AHS (autologous human seruml was used instead of 10% FCS (fetal calf serum) in the recombinant IL-2 cultures of inductive phase of LAK. This suppression was not only for the inductive phase of LAK activation but also for the cvtotoxic phase of LAK activity, because AHS could suppress the cytotoxic capability of LAK once Lhey were activated by recomhinant IL-2. The cytotoxic response of LAK of normal persons and stomach cancer patients cultured with nonnal seurm were significantly higher than the of LAK cultured with stomach cancer patient serum (P<0.001). The cytotoxic response of LAK which was cultured with stomach cancer patient's serum with 1 unit/ml of recombinant IL-2 were significantly higher than those of LAK which was cultured with stomach cancer patients serum with 1 unit/ml of recombinant IL-2 (P<0.001). So, the LAK activity wihich was supprevsed by strimach cancer patients serum could be rwercome by increasing thc conccntration of recomhinant IL-2 during the incluctive phase of LAK activation and the cytotoxic phase of LAK.
Key words: Limphokinc activated killer cells. LAK, Autologous human serum, AHS
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