CD3和CD28双刺激活化的健康人外周血T淋巴细胞与胃泌素拮抗剂合用对结肠癌细胞的杀伤作用

目的探讨联合应用胃泌素拮抗剂和CD3/CD28单克隆抗体共刺激活化健康人外周血T淋巴细胞(PBLs)在体外对结肠癌细胞株HT-29的影响及杀伤途径。为结直肠癌的过继免疫治疗提供参考。方法外周血T淋巴细胞(PBLs)的分离与体外培养;CD3/CD28共刺激活化外周血T淋巴细胞(PBLs);MTT法检测活化细胞的体外淋巴细胞毒作用,并用流式细胞仪检测结肠癌细胞的凋亡情况。结果用CD28共刺激细胞和CI—988分别处理6d后对结肠癌细胞株HT-29抑制率为66%和47.5%,二者合用后3d和6d抑制率分别为41%和90.5%;较单用任何一种处理对结肠癌细胞株HT-29杀伤作用更强(P<0.01)。流式细胞仪FCM图像可见实验组细胞群坏死比例为30.1%,凋亡细胞占19.2%,而对照组仅有坏死细胞0.22%,凋亡细胞1.6%。结论联合应用胃泌素拮抗剂CI-988可以提高单抗协同诱导的效应细胞对结肠癌细胞株HT-29总体抑制,而坏死细胞进一步增多说明效应细胞是通过诱导肿瘤细胞坏死及细胞凋亡两条途径来实现抑制作用。

Experimental study of adoptive immmunotherapy in colon cancer cell with peripheral blood lymphocytes induced by CD3 and CD28 when combined with gastrin receptor antagonist

Objective: To investigate the killing effect of costimulate activation of PBLs with CD28 and CD3 McAb on the human colorectal carcinoma cell lines when combined with gastrin receptor antagonist in vitro. Methods: PBLs were obtained by Ficoll-Hypeque density gradient centrifugation. The cancer cells were cultured with PBLs and CD28/ CD3 McAb and CI-988,then the growth curve of cells was obtained.The killing efficiency was measured by MTT method.The submicroscopic structure of cells was observed by electron microscope. Apoptosis was verified by a flow cytometer.Results: After treated with costimulate activation of PBLs with CD28 and CD3 or CI-988,the inhibition rate of HT-29 cells was 60.4%.When two reagents were used in combination,the inhibition rate was 90.5%. By MTT method,combination of costimulate activation of PBLs with CD28 and CD3 and Cl-988 possessed superior killing effect in comparison to single therapy (P<0.01). Flow cytometer verified that cells were apoptosized after exposed to costimulate activation of PBLs with CD28 and CD3 and CI-988. Conclusion: Gastrin receptor antagonist can elevate the killing effect of anti-CD28 stimulated cells on colorectal carcinoma. Apoptosis is possibly one of the reasons of the synergistic action. They killed the cells in two ways:necrosis and apoptosis.