IL-12对BALB/c小鼠Th17细胞的分化的影响

Detection of Thl7 cells in splenocytes of BALB/c mouse

目的:观察细胞因子IL-12对Th17细胞分化的影响。方法:小鼠脾淋巴细胞经抗CD3单克隆抗体(mAb)和不同浓度的重组小鼠IL-12刺激,3d后使用ELISA方法观察培养物上清液中IL-17的产生情况。并使用细胞内细胞因子染色的方法,通过流式细胞术观察CD3mAb和重组小鼠IL-12刺激对Th1和Th17细胞分化的影响。结果:Th17细胞不分泌IFN-γ、IL-5、IL-10等细胞因子,不表达Foxp3,是一个独立的细胞亚群。不同浓度的重组小鼠IL-12可以诱导抗CD3mAb的T细胞分泌IFN-γ,并向Th1细胞方向分化。同时,IL-12可以抑制活化的T细胞分泌IL-17,抑制T细胞向Th17细胞分化。结论:IL-12可以抑制Th17细胞的分化。

AIM： To observe the role of IL-12 in Th17 cells differentiation. METHODS： Mouse splenocytes were prepared and cultured with aCD3 alone or plus different concentration of recombined mice IL-12, 3 days later, IL-]7 in the culture supernatant was assayed by ELISA. In addition, Mouse splenocytes were stimulated byαCD3 with or without IL-12 for 48 h, then cell were washed, and restimulated by PMA and Ionomycin, the expression of intracellular IL-17 and IFN-y by CD4^＋ and CD8^＋ T cells were examined at single cell level by Flow Cytometric analysis. RESULTS： Th-17 cells is a independent cell population, which did not express IL-5, IL-10, IFN-y and Foxp3. IL-12 could induce active mouse T cells produce large amount of IFN-y and the differentiation toward Thl cell. On the contrary, IL-12 could inhabit the production of IL-17 and differentiation toward Th17 cells. CONCLUSION： IL-12 inhabit T cell differentiation toward Th17 cells.