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目的 :阐明重组减毒鼠伤寒沙门氏菌在体内早期感染抗原呈递细胞 (APC)的动态变化规律。方法 :用一定剂量的表达绿色荧光蛋白 (GFP)的重组减毒鼠伤寒沙门氏菌X4 5 5 0(pYAGFP)口服接种BALB/c小鼠。 3d后 ,取腹腔巨噬细胞培养 2 4h ,用荧光显微镜观察。另以该细菌静脉注射小鼠 ,于感染后 3、6和 12h,制备脾脏、肝脏低浮密度细胞。用流式细胞术检测巨噬细胞和树突状细胞的感染率。结果 :巨噬细胞感染X4 5 5 0 (pYAGFP)的百分率 ,腹腔中约为 5 0 % ,肝脏和脾脏中约为 2 0 %～ 4 0 %。树突状细胞感染X4 5 5 0 (pYAGFP)的百分率 ,脾脏中约为 4 %～ 10 % ,肝脏中约为 10 %～ 2 0 %。巨噬细胞的吞噬能力明显高于树突状细胞。结论 :感染早期 ,重组减毒鼠伤寒沙门氏菌在体内已被APC所摄取 ,为诱导有效的免疫应答提供了先决条件。
Objective: To clarify the dynamic changes of recombinant attenuated Salmonella typhimurium in early stage antigen-presenting cells (APC) in vivo. Methods: BALB / c mice were orally inoculated with a dose of recombinant attenuated Salmonella typhimurium X4 5 05 (pYAGFP) expressing green fluorescent protein (GFP). After 3d, macrophages were cultured for 24 hours and observed under fluorescence microscope. In addition, mice were injected intravenously with the bacterium, and spleens and low-density cells of the liver were prepared at 3, 6 and 12 h after infection. Flow cytometry was used to detect the infection rates of macrophages and dendritic cells. RESULTS: The percentage of X4 5 05 (pYAGFP) infected by macrophages was about 50% in the abdominal cavity and 20% -40% in the liver and spleen. The percentage of dendritic cells infected with X4 5 05 (pYAGFP) is about 4% -10% in the spleen and about 10% -20% in the liver. Macrophages phagocytosis was significantly higher than dendritic cells. CONCLUSION: In the early stages of infection, recombinant attenuated Salmonella typhimurium is ingested by APC in vivo and provides a prerequisite for inducing an effective immune response.