Effects of cryopreservation and phenylacetate on biological characters of adherent LAK cells from patients with hepatocellular carcinoma  

作　　者：Ning Zheng Sheng-Long Ye Rui-Xia Sun Yan Zhao Zhao-You Tang,Liver Cancer Institute and Zhongshan Hospital,Fudan University,Shanghai 200032,China 

出　　处：《World Journal of Gastroenterology》2002年第2期233-236,共4页世界胃肠病学杂志（英文版）

基　　金：the National 9th Five-Year Program of China,No.96-906-01-20

摘　　要：AIM: To improve the preparation of adherent lymphokine-activated killer (A-LAK) cells and to study the effects ofcryopreservation and phenylacetate (PA) on biologicalcharacters of A-LAK cells.METHODS: A-LAK cells were obtained from peripheral bloodmononuclear cells (PBMCs) of the patients withhepatocellular caminoma (HCC) by using L-phenylalaninemethyl ester ( PME ) to deplete immunosuppressivermonocytes. Proliferative activity of SMMC7721 cell line aftertreatment with phenylacetate (PA) was observed. A-LAKcells were treated with the supernatant of SMMC7721 cellsthat had been pretreated with PA. The changes ofproliferation, cytotoxicity and phenotype of A-LAK cellswere investigated after cryopreservstion.RESULTS: The expansion of A-LAK cells (96.79± 69.10 foldson Day 14 ) was significantly higher then that of non-adherent LAK (NA-LAK) cells (22.77 ± 13.20) as well asconventional LAK cells (4. 64 ± 0. 91 ). PA significantlysuppressed the growth of SMMC7721 cells, and the inhibitorratio was 46 %. rite supernatant of cultured tumor cellsintensively suppressed the proliferation and cytotoxicity ofA-LAK cells, but the suppressive effect of the supernatantwas previously decreased after treatment with PA.Impairments in proliferation and cytotoxicity of A-LAK cellsimmediately after thawing of cryopreservation and recoveryafter reincubation with IL-2 were observed. The cytotoxicltyof thawed A-LAK cells on Day 5 was significantly higher thanthat of fresh A-LAK before freezing (54.8 ± 10.2 % vs 40.5 ±6. 4 %). No significant change in the percentage oflymphocyte subsets was identified in frozen A-LAK cells ascompared with that in the fresh control cells.CONCLUSION: A-LAK cells can he simply prepared by usingPME, and showed a synergistic anti-tumor effect with thecombination of PA. Cryopreseration can increase theimmunoactivities of A-LAK cells from the patients withhepatocellular caminoma .AIM:To improve the preparation of adherent lymphokine- activated killer(A-LAK)cells and to study the effects of cryopreservation and phenylacetate(PA)on biological characters of A-LAK cells. METHODS:A-LAK cells were obtained from peripheral blood mo■onuclear cells(PBMCs)of the patients with hepatocellular carcinoma(HCC)by using L-phenylalanine methyl ester(PME)to deplete immunosuppressive monocytes.Proliferative activity of SMMC7721 cell line after treatment with phenylacetate(PA)was observed.A-LAK cells were treated with the supernatant of SMMC7721 cells that had been pretreated with PA.The changes of proliferation,cytotoxicity and phenotype of A-LAK cells were investigated after cryopreservation. RESULTS:The expansion of A-LAK cells(96.79±69.10 folds on Day 14)was significantly higher than that of non- adherent LAK(NA-LAK)cells(22.77±13.20)as well as conventional LAK cells(4.64±0.91).PA significantly suppressed the growth of SMMC7721 cells,and the inhibitor ratio was 46%.The supernatant of cultured tumor cells intensively suppressed the proliferation and cytotoxicity of A-LAK cells,but the suppressive effect of the supernatant was previously decreased after treatment with PA. Impairments in proliferation and cytotoxicity of A-LAK cells immediately after thawing of cryopreservation and recovery after reincubation with IL-2 were observed.The cytotoxicity of thawed A-LAK cells on Day 5 was significantly higher than that of fresh A-LAK before freezing(54.8±10.2 % vs 40.5± 6.4 %).No significant change in the percentage of lymphocyte subsets was identified in frozen A-LAK cells as compared with that in the fresh control cells.CONCLUSION: A-LAK cells can be simply prepared by using PME. and showed a synergistic anti-tumor effect with the combination of PA. Cryopreservation can increase the immunoactivities of A-LAK cells from the patients with hepatocellular carcinoma.

关 键 词：肝细胞癌 粘附淋巴因子活化杀伤细胞 冷冻保存 醋酸苯酯 

分 类 号：R735.7[医药卫生—肿瘤]