抗人卵巢癌×抗人CD3双特异单链抗体介导的效应细胞在体外对卵巢癌细胞的杀伤作用  

Cytotoxicity for a bispecific single-chain antibody against T cells and ovarian carcinoma

作　　者：赵瑞[1] 方敏[1] 李骅[1] 林晴[1] 郭蔼光[2] 薛菘[2] 黄华樑[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所 [2]西北农林科技大学生命科学学院

出　　处：《中华微生物学和免疫学杂志》2003年第6期476-479,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家 8 63高科技研究发展计划基金资助项目 ( 10 2 -0 9-0 4- 0 1)

摘　　要：目的　研究双特异单链抗体 (scBsAb)介导的Jurkat细胞 (CD3+ )及人外周血单个核细胞(PBMC)在体外对人卵巢癌细胞株SKOV3细胞的杀伤活性 ,从而探讨其用于卵巢癌治疗的可能性。方法以抗人卵巢癌×抗人CD3scBsAb激活效应细胞 ,以SKOV3为靶细胞 ,用MTT法测定不同实验条件下的杀伤活性。结果　 (1)以重组白细胞介素 2 (rIL 2 )、抗CD3单克隆抗体、抗CD2 8重链单域抗体 (VH)预刺激Jurkat及PBMC细胞比单纯用scBsAb激活效应细胞杀伤活性高 ;(2 )以Jurkat细胞或PBMC细胞作为杀伤细胞可得到相似的杀伤活性 ,最高杀伤率均在 75 %左右 ;(3)杀伤活性与scBsAb的浓度、作用时间及效靶比有关。对于Jurkat细胞 ,在抗体终浓度为 2 1μg ml,反应时间为 4 8h ,效靶比为 10∶1时达到最大杀伤率 74 .8% ;对于PBMC细胞 ,在抗体浓度为 2 0 μg ml,反应时间为 72h ,效靶比为 1∶1时达到最大杀伤率73.1%。 (4 )多因子联合应用能有效提高杀伤活性。结论　scBsAb在体外能够有效介导效应细胞杀伤肿瘤细胞 ,有明显的抗癌作用 ,具有潜在的应用前景。Objective To investigate the in vitro effect of bispecific single-chain antibody-mediated tumor cells killing by Jurkat cells and PBMC cells. Methods Jurkat cells or PBMC cells of healthy individuals were co-cultured with rIL-2, anti-CD3 monoconal antibody(McAb) and anti-CD28 scFv (VH). SKOV3 target cells and different values of scBsAb were plated in wells of the 96-well flat-bottom plates and incubated overnight. Effector cells were added into the tumor cell monolayers and plates were incubated for 4?h, 20?h, 24?h, 48?h and 72?h. Cytotoxic activity was evaluated by MTT assay. Results (1) Cytotoxicity activities of effect cells stimulated by a certain values of rIL-2, anti-CD3 McAb and anti-CD28 scFv (VH) were stronger than those stimulated with scBsAb alone. (2) The results showed that under the existing of scBsAb, Jurkat cells and PBMC cells had the similar effect on tumor cells. The cytotoxicities were respectively up to 74.8% and 73.1%. (3) The cytotoxic activities relied on the values of scBsAb, time, and the ratio of effect cells to the tumor cells. For Jurkat cells, 21μg/ml scBsAb, 48?h and E/T=10∶1 would be proper, the cytotoxicity was up to 74.8%; For PBMC cells, 20μg/ml scBsAb, 72?h and E/T=1∶1 would be an ideal condition, the cytotoxicity was up to 73.1%. (4) It was a good strategy of using combined cytokines. Conclusion scBsAb would be a promising immunological tools for the elimination of tumor cells.

关 键 词：卵巢癌 双特异单链抗体 外周血单个核细胞 CD3+细胞 

分 类 号：R737.31[医药卫生—肿瘤]