小鼠阴道黏膜接种布鲁菌S_2疫苗株树突状细胞动力学变化

将25只小鼠随机分为5组,1~4组经阴道接种布鲁菌,第5组阴道滴注PBS(对照组),分别在处理后12、24、487、2 h摘取髂内淋巴结,用S-100抗体进行免疫组织化学染色,观察小鼠树突状细胞(Dendritic cells,DCs)在淋巴结的分布变化;同时收集外周血用ELISA法进行IFN-γ检测。用布鲁菌负载DCs,分别在2、4、6、8、10、12 h收集细胞悬液制备涂片,姬姆萨染色。结果显示:阴道接种布鲁氏菌疫苗12~48 h髂内淋巴结中DCs的数量明显增多,成群分布,呈现由浅层皮质向副皮质区迁移的趋势;72 h后大量DCs呈单在分布,分布范围增大;接种疫苗后血清中IFN-γ含量逐渐升高,至48 h,与对照组相比差异显著(P0.05),72 h后,血清中IFN-γ含量达到最大值,与对照组相比差异极显著(P0.01);布鲁菌负载DCs后,在4 h仅有少量DCs吞噬布鲁菌,至12 h吞噬布鲁菌的DCs数量明显增加。结果表明,DCs具有较强捕获布鲁菌的能力,并在抗原刺激下向引流淋巴结迁移,刺激淋巴结免疫细胞产生IFN-γ应答。

Dynamics of dendritic cells in vaginal mucosa of mice inoculated with Brucella suis

Twenty-five mice were randomly devided into five groups of five mice each. Mice of four groups were inocu-lated by intravaginal administration of Brucella vaccine. The iliac lymph nodes (ILNs) were removed pose-treatment 12,24,48,and 72 h,respectively. As a control,the mice in fifth group were intravaginally instillated with phosphate-buffered saline (PBS) and ILNs were harvested. ILNs were processed routinely to obtain paraffin-embedded sec-tions. Polyclonal rabbit.anti-S100 antibody was used to detect dendritic cells(DCs) in ILNs. Peripherial blood samples were collected for ELISA assay of IFN-γ. DCs were pulsed with the Brucella vaccine in vitro and stained with Giem-sa. It was shown, that the numbers and distribution pattern of DCs changed dramatically in ILNs after inoculation with Brucella vaccine. The numbers of DCs in cortex of ILNs increased and concentrated in cluster from 12-48 h,and migrated from the outer cortex into the paracortical zone. The distributing area of DCs enlarged at 72 h. The level of IFN-γ in serum increased, significantly compared with control at 48 h(P<0.05) and reached the peak level pose-treatment 72 h with significantly higher than that of control group(P<0.01). DCs only captured a small amount of Brucella pose-treatment 4 h in vitro. The numbers of DCs captured Brucella increased constantly and reached the peak level at 12 h. It is concluded that DCs have a powerful capability to capture Brucella. The antigen-loaded DCs in vagina arc capable of rapidly migrating to the draining lymph nodes,indicating that DCs in vagianl mucosa is of Bru-celia antigen presenting cells and initiate immune response characteristized by of release of IFN-γ.