双特异抗体CD3xAFP介导CIK细胞对AFP阳性肝癌细胞的杀伤应用

目的建立双特异抗体anti-CD3xanti-AFP介导CIK细胞对AFP阳性肝癌细胞的特异杀伤。方法利用anti-CD3抗体联合细胞因子IL-2活化人外周血淋巴细胞扩增CIK细胞,流式细胞仪分析细胞表型。柠檬酸三钠还原法制备金溶胶,物理吸附制备载荷anti-CD3和anti-AFP两种单抗的胶体金制剂,光镜检测抗体活性。AlarmaBlue法检测双特异抗体介导CIK细胞对肝癌细胞HepG-2和宫颈癌细胞Hela的杀伤活性。结果 14 d活化后的人外周血淋巴细胞鉴定为CIK细胞。抗体活性检测证实纳米金制剂保持了anti-CD3xanti-AFP的双特异抗体活性,在其介导下,CIK对特异靶细胞HepG-2杀伤效率为42%,提高11.4%(P<0.05),而对非特异靶细胞Hela杀伤无明显变化。结论金标双特异抗体anti-CD3xanti-AFP可以加强CIK细胞对AFP阳性肝癌细胞的杀伤作用。

Cytotoxicity of cytokine-induced killer cells targeted by the bispecific antibody anti-CD3xanti-AFP on AFP positive hepatoma cells

Purpose To establish the method in which CIK cells targeted by the bispecific antibody anti-CD3xanti-AFP specifically lyse AFP positive hepatoma cells.Methods CIK cells were activated by anti-CD3 antibody and IL-2 from human peripheral blood lymphocytes and cell phenotypes were detected by flow cytometer.Gold colloid was prepared by the method of citric acid tri-sodium revert,bispecific colloidal gold antibody anti-CD3xanti-AFP were prepared by physical adsorption,and activities of antibodies were detected by light microscopy.Cytotoxic activities of CIK cells on hepatoma cells HepG-2 and cervical carcinoma cells Hela were tested by Alarma Blue assay.Results Human peripheral blood lymphocytes after 14 days′ activation were characterized as CIK cells.Antibody activity detection showed that the colloidal gold preparation kept activities of the bispecific antibody anti-CD3xanti-AFP.Targeted by the bispecific antibody,the efficiency of CIK lysing to HepG-2 cells was 42.0%,increased by 11.4%.The efficiency of CIK lysing to Hela cells showed no significant increase.Conclusion CIK cells targeted by the bispecific colloid gold antibody anti-CD3xanti-AFP could lyse AFP positive hepatoma cells better.