TLR激动剂抑制IL-27诱导的THP-1细胞HLA-DR的表达

TLR Agonists Inhibit IL-27-Induced HLA-DR Expression in THP-1 Cells

目的研究白介素27(IL-27)对THP-1单核细胞系Ⅱ类转录活化子(classⅡtransactivator,CⅡTA)和Ⅱ类主要组织相容性复合物(major histocom patibility complex classⅡ,MHCⅡ)分子的表达的影响及Toll样受体(Toll-likereceptor,TLR)激动剂LPS和Pam3CSK4的干预作用。方法RT-PCR检测CⅡTAⅠ、Ⅲ和Ⅳ以及人白细胞抗原(human leukocyte antigen,HLA)-DRA和干扰素调节因子-1(interferon regulatory factor-1,IRF-1)mRNA的表达。半定量PCR检测HLA-DRA、DRB、DPA、DPB、DQA、DQB、IRF-1、CⅡTA mRNA的表达。流式细胞术检测细胞表面HLA-DR的表达。结果IL-27刺激24h后可分别上调THP-1细胞CⅡTAⅢ、CⅡTAⅣ和IRF-1mRNA表达水平。IL-27刺激24h和48h可升高MHCⅡ类分子mRNA的表达,并能诱导HLA-DR在28%和53%的THP-1细胞表面表达。在THP-1细胞和PMA诱导的THP-1巨噬细胞,LPS和Pam3CSK4均可抑制IL-27诱导的CⅡTA和HLA-DR的表达。结论IL-27可上调THP-1细胞CⅡTA和MHCⅡ类分子的表达,这种作用可被LPS和Pam3CSK4抑制。

Objective To investigate the effect of interleukin-27 （IL-27） on the expression of class Ⅱ transactivator （CIITA） and major histocompatibility complex class Ⅱ （MHC Ⅱ） genes in THP-1 cells and the intervention by Toll-like receptor （ TLR ） agonists LPS and Pam3CSK4. Methods Real-time PCR was performed to examine the mRNA expression levels of CI- ITA Ⅰ , Ⅲ and Ⅳ, human leukocyte antigen （HLA） -DRA and interferon regulatory factor-1 （IRF-1） . Semi-quantitative RT-PCR was performed to assess the mRNA levels of HLA-DRA, DRB, DPA, DPB, DQA, DQB, IRF-1 and CIITA. Flow cytometry was used to determine the surface expression of HLA-DR molecules. Results After 24 hours of stimulation, IL-27 increased the CIITA m, CIITAIV and IRF-1 mRNA levels 14-, 38- and 8-folds respectively. IL-27 increased MHC II mRNA levels and induced surface expression of HLA-DR in 28% and 53% of THP-1 cells after 24-hour and 48-hour of incubation. In both THP-1 cells and PMA-activated THP-1 macrophages, LPS and Pam3CSK4 inhibited IL-27 inducible CIITA and MHC Ⅱ expression. Conclusion IL- 27 could increase CIITA and MHC Ⅱ expression in THP-1 cells, and these effects could be inhibited by TLR2 and TLR4 agonists.