Ureaplasma urealyticum GrpE promotes mouse dendritic cell maturation and elicits Th1 immune responses
10.3760/cma.j.cn112309-20210525-00175
- VernacularTitle:解脲脲原体GrpE蛋白促进小鼠树突状细胞成熟并诱导Th1型免疫应答
- Author:
Fangyi GUO
1
;
Yanhong TANG
;
Hongxia YUAN
;
Wenjun ZHANG
;
Jing XIANG
;
Pengqin LIU
;
Wenyou TENG
;
Ranhui LI
;
Guozhi DAI
Author Information
1. 南华大学衡阳医学院附属郴州医院/郴州市第一人民医院检验医学中心，郴州 423000
- Keywords:
Ureaplasma urealyticum;
GrpE;
Dendritic cell;
Immune response
- From:
Chinese Journal of Microbiology and Immunology
2022;42(1):41-49
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of Ureaplasma urealyticum GrpE ( Uu-GrpE) on the maturation of dendritic cells and the polarization of T cells. Methods:Uu-GrpE was expressed and purified, and then identified by Western blot. The cytotoxicity of Uu-GrpE to mouse bone marrow-derived dendritic cells (BMDCs) was analyzed by LDH kit. After stimulating BMDCs with Uu-GrpE, the expression of costimulatory molecules, CD80, CD86 and major histocompatibility complex Ⅱ (MHCⅡ), on the surface of BMDCs was detected by flow cytometry, and ELISA was used to detect the cytokines such as IL-12p70, TNF-α, IL-1β and IL-6. CD4 + Na?ve T cells were isolated from mouse spleen tissues by magnetic beads. A co-culture system of BMDCs and Na?ve T cells was constructed to analyze the effects of GrpE-stimulated mature BMDCs (GrpE-BMDCs) on T cell proliferation and polarization towards Th1/Th2. Mice were immunized with GrpE-BMDCs through the tail vein, and the induced humoral and cellular immune responses were detected by ELISA and flow cytometry. Results:Uu-GrpE was successfully express and high purity BMDCs were isolated. Uu-GrpE could stimulate BMDCs to secrete cytokines such as IL-12p70, TNF-α, IL-1β and IL-6 without having cytotoxicity. Uu-GrpE significantly increased the expression of CD80 [mean flourscence indensity (MFI): (324.00±22.11) vs (91.03±10.95), P<0.01], CD86 [MFI: (1 176.00±51.39) vs (217.00±14.93), P<0.01] and MHCⅡ [MFI: (708.70±56.32) vs (185.70±16.77), P<0.01] on BMDCs. Compared to the GrpE-BMDCs only group and GrpE (boiled)-BMDCs+ T cell group, the GrpE-BMDCs+ T cell group showed significantly increased T cell proliferation [stimulation index: (7.25±0.21) vs(6.55±0.23) and (6.09±0.35), both P<0.05], and dramatically promoted T cell secretion of IL-2 and IFN-γ [IL-2: (145.60±14.67) pg/ml vs(55.92±3.12) pg/ml and (26.05±2.40) pg/ml, P<0.05 and P<0.01; IFN-γ: (267.20±37.80) pg/ml vs(146.70±20.65) pg/ml and(27.84±6.69) pg/ml, both P<0.05]. However, no significant change was observed in the expression of Th2-type cytokines. Moreover, the adoptive transfer of GrpE-BMDCs induced a Th1-type immune response. Conclusions:Uu-GrpE could stimulate the maturation and polarization of BMDCs. Moreover, it could induce Th1 immune response as a candidate protein vaccine for Ureaplasma urealyticum.
