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目的研究猪布鲁菌S2株感染后死亡的巨噬细胞在启动抗布鲁菌感染免疫应答中的作用。方法以猪布鲁菌S2株体外感染小鼠腹腔巨噬细胞,感染1 h后,饥饿培养5 d致细胞死亡;细胞碎片与骨髓源性树突状细胞(DC)共培养24、48、72 h,ELISA检测细胞上清中白细胞介素12(IL-12)、肿瘤坏死因子α(TNF-α);以S2株体外感染羧基荧光黄二乙酸盐琥珀酰亚胺酯(CFSE)标记的小鼠巨噬细胞,饥饿避光培养,将细胞碎片与藻红蛋白标记的DC避光共培养1 h后,激光共聚焦显微镜检测DC摄取巨噬细胞碎片情况;以S2株感染后死亡的巨噬细胞碎片接种于BALB/c小鼠腹腔,1周后加强免疫1次;加强免疫3 d后,ELISA检测血清中IL-4、IL-2和γ干扰素(IFN-γ)水平。结果负载S2株感染后死亡巨噬细胞碎片的DC TNF-α的分泌水平明显高于未感染组,但不分泌IL-12;且S2株感染后死亡的巨噬细胞能被DC摄取到细胞内;接种S2株感染后死亡的巨噬细胞的小鼠血清中IL-2、IL-4和IFN-γ水平明显高于对照组。结论感染猪布鲁菌S2株死亡的巨噬细胞可激活DC、使其提呈抗原、并诱导机体产生抗感染免疫。
Objective To study the role of macrophages that died after infection with swine brucellosis S2 in the initiation of anti-Brucella infection. Methods In vitro infection of murine peritoneal macrophages with porcine brucellosis S2 strain was performed. After 1 h of infection, the cells were killed after starvation for 5 days. The cells were co-cultured with bone marrow-derived dendritic cells (DCs) 24, 48, 72 h, IL-12 and TNF-α in the supernatant of the cells were detected by ELISA. S2 strain was infected with carboxyfluorescein diacetate succinimidyl ester (CFSE) in vitro Macrophages were cultured in starvation and darkness. The cells were co-cultured with phycoerythrin-labeled DCs for 1 h. The macrophages were collected by laser confocal microscopy. The fragments were inoculated into the peritoneal cavity of BALB / c mice and boosted one time after one week. The serum levels of IL-4, IL-2 and IFN-γ were detected by ELISA 3 days after boosting. Results The level of DC TNF-α secreted by dead macrophages loaded with S2 strain was significantly higher than that of non-infected cells but not secreted IL-12. The macrophages that died after S2 infection could be ingested by DC The levels of IL-2, IL-4 and IFN-γ in the sera of mice that were killed after inoculation of S2 strain were significantly higher than that of the control group. Conclusion Macrophages infected with Brucella spp. Strain S2 can activate DCs to produce antigens and induce the body to produce anti-infective immunity.