CD_3单抗、CD_(28)/CpGODN共刺激活化的PBMC对膀胱癌细胞杀伤作用的研究

Experimental Study of Adoptive Immmunotherapy in Bladder Cancer Cell With Peripheral Blood Mononuclearl (PBMC) Induced by CD_3 McAb and CD_(28)/CpGODN

目的 探讨CD3单克隆抗体与CD2 8/CpGODN共刺激活化PBMC在体外对膀胱癌细胞株T2 4 及Scarber杀伤强度及杀伤作用机理。为膀胱癌的过继免疫治疗提供新的效应细胞。方法 MTT法检测活化细胞的体外淋巴细胞毒作用 ,并用电镜观察效应细胞杀伤膀胱癌细胞的超微结构。结果 CD2 8共刺激细胞对T2 4 杀伤作用较强 ,达到 69.35 %± 1 .1 7% ,而CpGODN诱导的活化细胞对Scarber杀伤较强 ,达到 63.1 1 %± 2 .0 3%。且效应细胞 :靶细胞均需达到 2 0 :1才达到半数杀伤作用。电镜结果显示 ,效应细胞作用 6小时肿瘤细胞就大部分发生坏死 ,部分肿瘤细胞可见凋亡 ,说明效应细胞是通过诱导肿瘤细胞坏死及凋亡实现杀伤作用的。结论 CD2 8单抗协同诱导的效应细胞对T2 4 杀伤效果较好 ,而CpGODN刺激细胞对Scarber杀伤较强 ,活化的淋巴细胞杀伤膀胱癌细胞是通过坏死及凋亡两条途径来实现的。

Objective To investigate a new clinical therapeutic approach for adoptive immmunotherapy in bladder cancer and its killing principle. Methods PBMC were obtained by Ficoll-Hypeque density gradient centrifugation. In experiment,the test groups were cultured with CD 3McAb and diffferent dilution of CD 28 McAb or CpGODN.To assay the lymphocytes' cytotoxicity with MTT methods, the microchange of the tumor cells was investigated by electron microscope. Results Anti-CD 28 stimulated cells could kill T 24 bladder cancer cells more effectively than other activated cells, CpGODN stimulated cells could kill Scarber bladder cancer cells more stronger. Only effector:target to 20:1 could get effective cytotoxicxicity, the result of electron microscope showed that large proportion of target cells necrosis rapidly when cultured with effectors and partial target cells apopotise. Conclusion Anti-CD 28 -costimulated cells could kill T 24 bladder cancer cells effectively.CpGODN stimulated cells could kill Scarber bladder cancer cells stronger, they killed tar- cells in two ways:necrosis and apopotise.