CD_3McAb、CD_28McAb、CpG ODN刺激PBMC活化的研究

PBMC Proliferation and Activation Stimulated by CD_3McAb, CD_(28) and CpG ODN

目的 为肿瘤生物治疗寻找高效、安全的效应细胞。方法 采用CD3 单克隆抗体与CD2 8单克隆抗体及CpGODN共刺激外周血单核细胞活化增殖。对增殖细胞的增殖量、剂量依赖性与细胞表型进行测定。结果 单用CD3 单克隆抗体和小剂量的IL - 2就可以引起PBMC有效扩增 ,CD2 8单抗可提供协同刺激信号 ,使PBMC扩增达到最高。而CpGODN亦可提供第二信号 ,但较CD2 8单抗为弱。CD2 8协同扩增呈剂量非线性相关型 ,而CpGODN则为剂量正相关。表型测定表明共刺激细胞中NK细胞比率明显增高 ,分别达到 33.1 0 %± 1 3.82 % (CD3 +CD2 8) ,2 9.5 0 %± 1 3.2 0 % (CD3 +CPGODN) ,与对照组比较差异有显著性 (P <0 .0 5 )。同时 ,CD4+ CD8+ T细胞比值发生倒置。结论 CD2 8、CpGODN均能协同CD3 单抗诱导PBMC活化增殖 ,CD2 8单抗激活无剂量线性相关 ,而CpGODN激活有剂量正相关。激活的细胞是以NK细胞、CD4+ CD8-、CD4- CD8+ T细胞为主体的异质细胞群。

Objective PBMC proliferation and activation stimulated by CD 3McAb , CD 28 /CpG ODN. assess the effective of CD 3-McAb and CD 28 /CpG ODN to induce PBMC proliferation, and analyse proliferational cell type. Methods PBMC were cultured with CD 3McAb and different dilution of CD 28 McAb or CpG ODN,To assay the activated lymphocytes′ proliferation use MTT methods. Activated cells were analyzed by flow cytometry, including CD 16 +CD 56 + NK cells; CD 4 +/CD 8 -, CD 4 -/CD 8 + T cells. Results Single CD 3McAb combined with low-dose IL-2 could induced PBMC proliferation, CD 28 McAb could transmited stimulational signals. So that PBMC could get maximal response ,in the 5th day ; CpG ODN could serve as immunostimulation. But intensity was lower than CD 28 mAb. Anti-CD 28 -stimulated cells grow in a dose-independent manner, but CpG ODN was dose-dependent. According to the phenotypes results, we found that the content of NK cells increased evidently,significant higer than control groups.we also found that CD4 +/CD8 +T cells decreased. Conclusion Both CD 28 mAb and CpG ODN combined with CD 3mAb could induced PBMC proliferation, and the activated cells included NK cells , CD 4 +/CD 8 -,CD 4 -/CD 8 +T cells.