结核特异性多肽E6、E7和C14对单核-巨噬细胞亚型极化的影响

The influence of the Mycobacterium tuberculosis(MTB)-specific peptides E6, E7 and C14 on the monocyte-macrophage polarization

目的运用流式细胞术检测单核-巨噬细胞极化亚型M1(CD14^+CD16^+)及M2(CD14^+CD163^+),探讨结核特异性多肽对单核-巨噬细胞极化分型的影响。方法以3种结核特异性多肽E6、E7和C14作为刺激物,刺激人急性白血病单核细胞株(THP-1细胞)、结核病患者胸水单个核细胞及外周血单个核细胞(PBMC),用流式细胞术检测刺激物不同时间点对单核-巨噬细胞极化表面标记表达的影响。结果结核特异性多肽E6刺激THP-1 24+0 h、24+24 h、24+48 h、24+72 h后,CD16/CD163阳性率分别为16.85%/13.78%、19.59%/15.68%、18.14%/14.19%、13.61%/11.47%。E7刺激效果与E6相似,C14对THP-1的CD16/CD163表达影响不如前两种多肽强。结核特异性多肽E6刺激结核患者胸水或血标本单核细胞19 h后,CD14+CD16+阳性率(1#患者:1.66%,2#患者:4.37%)与CD14^+CD163^+阳性率(1#患者:1.76%,2#患者:2.82%)差异不大,但CD14^+CD16^+CD86^+阳性率(1#患者:2.20%,2#患者:6.16%)大于CD14^+CD163^+CD206^+阳性率(1#患者:1.37%,2#患者:0.92%)。E7刺激效果与E6相似,C14对结核患者胸水或血标本单核细胞CD14/CD16/CD163表达影响不如前两种多肽强。结论 3种结核特异性多肽特别是E6、E7体外刺激单核细胞株THP-1、结核患者胸水或血标本单核细胞主要向M1型单核-巨噬细胞极化。

Objective To explore the influence of Mycobacterium tuberculosis（MTB）-specific peptide to monocyte-macrophage polarization by utilizing flow cytometry to investigate the expression of CD14/CD16/CD163 of the cells. Methods The expressions of M1（CD14~＋CD16~＋）/M2（CD14~＋CD163＋） of the monocytes（THP-1, and the monocytes from the hydrothorax and peripherial blood from tuberculosis（TB） patients） were detected by flow cytometry with 3 MTB-specific peptides as stimulants at the different time. Results The positive percentages of CD16/CD163 of THP-1 cell line treated with the MTB peptide E6 at the times 24 ＋ 0 h, 24 ＋ 24 h, 24 ＋ 48 h, and 24 ＋ 72 h were 16.85%/13.78%, 19.59%/15.68%, 18.14%/14.19%, 13.61%/11.47%, respectively. The MTB peptide E7 exerted effect similar to E6, but the MTB peptide C14 showed a less effect. After treated with E6 at 19 h, the positive percentages of CD14~＋CD16~＋ of the monocytes of hydrothorax from 2 TB patients were 1.66%（No. 1 patient） and 4.37%（No. 2 patient）, as well as CD14~＋CD163~＋ were 1.76%（No. 1 patient） and 2.82%（No. 2 patient）. The difference between the two types of cells was not very remarkable, but the difference seemed to be significant when added another detection mark CD86 to CD14~＋CD16~＋, and CD206 to CD14~＋CD163~＋, respectively. E7 and C14 had the similar corresponding effects on the polarization of monocytes of hydrothorax and peripherial blood as observed at THP-1 cell line. Conclusion 3 MTB-specific peptides, especially E6 and E7, can mainly induce THP-1 and the monocytes of hydrothorax and peripherial blood from TB patients towards the M1 macrophage polarization.