【摘要】 目的　观察重组人可溶性CD4 0 配体 (rhsCD4 0 L)及CD4 0 L基因转染细胞 (CD4 0 L TC)对恶性B淋巴细胞体外生物学行为的影响 ,探讨rhsCD4 0 L在肿瘤生物治疗中的可能作用。方法　利用基因工程技术获得了rhsCD4 0 L和CD4 0 L TC ,分别与恶性B淋巴细胞株XG2、XG7、U2 6 6、82 2 6、Raji及Daudi共同作用 ,分析了CD4 0 激发对肿瘤细胞的体外增殖 (锥虫蓝计数法 )、细胞周期 (碘化丙锭掺入法 )、细胞表面共刺激分子的表达 (免疫荧光标记法 )以及细胞凋亡 (Annexin Ⅴ FITC法 )的效应。结果　①恶性B淋巴细胞株CD4 0 表达呈异质性 ,XG2高表达CD4 0 ,82 6 6、Raji和Daudi中度表达 ,而XG7和U2 6 6不表达CD4 0 。显微镜下观察发现 ,rhsCD4 0 L(5 μg ml)可引起XG2或Daudi细胞的同型聚集 ,该效应在作用 6～ 8h后即可出现 ;与CD4 0 L TC细胞共育后 (肿瘤细胞 :CD4 0 L TC =5∶1) ,XG2 ,Raji和Daudi细胞可粘附于CD4 0 L TC表面 ;②rhsCD4 0 L和CD4 0 L TC均可显著抑制XG2、Raji和Daudi细胞的体外增殖 ,导致XG2细胞呈现G1 期阻滞 ,而Raji和Daudi细胞阻滞于G2 期。并诱导XG2、Raji、Daudi细胞的凋亡 ,凋亡率分别为 :XG2细胞 2 3 3%和 18.8% ,Raji细胞 11.6 %和 8.9% ,Daudi 14 .2 %和 15 .9% ;③表型分析显示 :rhsCD4更多还原

【Abstract】 Objective To explore the effect of recombinant human soluble CD 40 ligand(rhsCD 40 L) and CD 40 L cDNA transfected cell (CD 40 L TC) on the behavior of malignant B lymphocytes, and investigate the possibility of using rhsCD 40 L as a new bio factor in tumor immunotherapy. Method rhsCD 40 L and CD 40 L TC were obtained by gene recombinant techniques. Multiple myeloma cell lines, XG2, XG7, U266 and 8226, B lymphoma cell lines, Raji and Daudi were selected to detect responses to rhsCD 40 L and CD 40 L TC stimulation. Cell growth curve, cell cycle, early apoptosis as well as membrane surface molecules on these cell lines were analyzed. Results ① The expression levels of CD 40 molecule on malignant B lymphocytes showed heterogeneity. High level of CD 40 on XG2, moderate on 8266, Raji, and Daudi, and no expression on U266 and XG7 were detected. The rhsCD 40 L stimulation gave rise to a typical homo type cell aggregation of XG2 and Daudi. Meanwhile, at least 10 to 20 of CD 40 + XG2 or CD 40 + Daudi cells were found adherent to one pre treated CD 40 L TC. ②Co incubation with rhsCD 40 L(5?μg/ml), or CD 40 L TC(tumor cell∶CD 40 =5∶1) resulted in a significant inhibition of in vitro cell growth of XG2, Raji and Daudi, with G 1 phase arrest for XG2 and G 2 phase for Raji and Daudi. These two kinds of CD 40 stimulators induced XG2, Raji and Daudi cells to apoptosis in vitro. The apoptotic rate for XG2 was 23.3%(rhs CD 40 L ) and 18.8%(CD 40 L TC), for Daudi 14.2% and 15.9%, and for Raji 11.6% and 8.9% respectively. ③Phenotype analysis showed that CD 95 expression levels were significantly up regulated on XG2, Raji and Daudi after stimulation with rhsCD 40 L or CD 40 L TC, and CD 80 and CD 18 expression levels on Raji were respectively enhanced and decreased. Conclusion The abilities to directly inhibit XG2, Daudi and Raji cell proliferation, to induce themapoptosis, as well as to up regulate immune co stimulator molecule CD 80 expression on Raji cells would make rhsCD 40 L a potentialbio factor for tumor immuno therapy.更多还原