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目的比较猪布鲁菌S2菌株感染巨噬细胞和树突状细胞(DC)的特点。方法采用瑞氏-吉姆萨染色法观察细胞形态并计算吞噬率;用ELISA法测定细胞培养上清中的IL-12和TNF-α以及与T细胞共培养上清中IFN-γ和IL-4的含量;用annexin-Ⅴ-FITC/PI双染,流式细胞仪检测细胞凋亡率。结果感染S2菌株1 h后巨噬细胞的吞噬率为(43.6±4.8)%,显著高于DC的吞噬率(13.08±2.36)%(P<0.05);正常巨噬细胞凋亡率为(3.09±1.21)%感染S2菌株后6、12和24 h的凋亡率分别为(19.89±1.36)%、(22.73±2.21)%和(42.44±3.12)%,明显高于DC感染后相同时间点的凋亡率(P<0.05),正常DC凋亡率为(1.82±0.01)%,DC感染S2菌株6、12、24 h凋亡率分别为(3.76±0.13)%、(7.87±0.56)%和(9.08±0.23)%。巨噬细胞在感染S2菌株后24、48 h分泌的IL-12均明显低于DC分泌IL-12的量(P<0.05);24、48、72 h分泌的TNF-α量均明显低于DC(P<0.01);在24、48、72 h与T细胞共培养分泌的IFN-γ含量均明显低于DC(P<0.01)。结论巨噬细胞吞噬猪布鲁菌S2菌株的能力高于DC,猪布鲁菌S2感染后巨噬细胞的凋亡率高于DC,感染S2菌株后DC的活化及提呈抗原并激活T细胞的能力强于巨噬细胞。
Objective To compare the characteristics of porcine Brucella S2 strains infected with macrophages and dendritic cells (DCs). Methods Wright-Giemsa staining was used to observe the cell morphology and the phagocytosis rate was calculated. The levels of IL-12 and TNF-α in cell culture supernatants and the levels of IFN-γ and IL-4 . The apoptosis rates were detected by annexin-Ⅴ-FITC / PI double staining and flow cytometry. Results Phagocytosis of macrophages was (43.6 ± 4.8)% after phage S2 infection for 1 h, significantly higher than that of DC (13.08 ± 2.36)% (P <0.05). The apoptosis rate of normal macrophages was (3.09 ± 1.21%. The apoptotic rates of S2, 6, 12 and 24 h after infected with S2 were (19.89 ± 1.36)%, (22.73 ± 2.21)% and (42.44 ± 3.12)%, respectively, (1.82 ± 0.01)%, respectively. The apoptotic rates of S2 infected DCs at 6, 12 and 24 h were (3.76 ± 0.13)% and (7.87 ± 0.56)%, respectively % And (9.08 ± 0.23)%. The levels of IL-12 secreted by macrophages 24 and 48 h after S2 infection were significantly lower than those secreted by DC (P <0.05), and the levels of TNF-α secreted by 24 and 48 h were significantly lower than DC (P <0.01). The levels of IFN-γ secreted by T cells co-cultured with T cells at 24,48,72 h were significantly lower than that of DCs (P <0.01). Conclusions Macrophages are more capable of phagocytizing the strain of Brucella ssp. S2 than DC. The apoptosis rate of macrophages after infection with porcine Brucella S2 is higher than that of DC. The activation of DC after S2 infection and antigen presentation and T cell activation More capable than macrophages.