hIL-10基因修饰的骨髓间充质干细胞的体外免疫抑制作用

An experimental study on the role of hIL-10 modified mesenchymal stem cell in heterologous T cell proliferation

目的研究hIL-10基因修饰的骨髓间充质干细胞(mesenchymal stam cell,MSC)对异种混合T淋巴细胞增殖的影响。方法反转录PCR克隆hIL-10基因并构建慢病毒hIL-10/pLOX-CWgfp,然后将hIL-10/pLOX-CWgfp转入豚鼠MSC内;ELISA法测定hIL-10基因修饰的MSC(hIL-10-MSC)培养上清液内hIL-10含量;CCK-8法检测hIL-10-MSC和其培养上清液对体外混合淋巴细胞培养的影响。结果hIL-10-MSC培养上清液的hIL-10水平明显高于未转染hIL-10者(P<0.05);hIL-10-MSC能抑制异种T淋巴细胞混合培养的增殖反应(P<0.05);加入IL-2能逆转MSC的抑制作用(P<0.05);hIL-10-MSC培养上清液能抑制异种T淋巴细胞混合培养的增殖反应(P<0.05)。结论hIL-10-MSC和其培养上清液对混合异种淋巴细胞培养有显著的抑制作用。

Objective To observe the influence of hlL-10 modified mesenchymal stem cells （ MSC ） on heterologous T cell proliferation. Methods hIL-10 was cloned by reverse transcription PCR and was used to construct hlL-10/pLOX-CWgfp. The lentivirus was transfected into cavy MSC. hiL- 10- MSC supernatant was gathered to detect IL-10 protein by ELISA assay, hIL-10-MSC or its medium effect on heterologous T cell proliferation were detected by CCK-8. Results IL-10 protein in the hiL- 10- MSC supernatant was markedly increased （P 〈 0.05 ）. hIL-10-MSC significantly inhibited heterologous T cell proliferation （ P 〈 0.05 ）, but did not induce T cell proliferation （ P 〈 0.05 ）. hIL- 10- MSC media performed the same effect （ P 〈 0.05）. IL- 2 reversed the inhibition effect （ P 〈 0.05 ）. Conclusion hlL-10-MSC may suppress heterologous T cell proliferation.