双价痢疾菌苗滴鼻免疫小鼠后不同部位淋巴细胞表型的变化

The observation of the lymphocyte pheno- type change in different lymphoid tissues of mice after intranasal immunization with bivalent Shigella vaccines

目的:观测双价痢疾工程菌苗滴鼻免疫小鼠后,不同时间、不同部位淋巴组织细胞表型的变化,探讨痢疾菌苗滴鼻免疫对黏膜和系统免疫应答的影响。方法:BALB/c小鼠随机分为3组,每组30只,分别以FSM-2117和FS-5416痢疾菌苗经滴鼻途径免疫小鼠4次,菌量依次为5×106、1×107、4×107和4×107CFU/只,对照组给予PBS,间隔2 wk。4次免疫后7、30和90 d活杀,分离NALT、鼻通道、脾、小肠PP结淋巴细胞,采用流式细胞术检测其淋巴细胞表型的变化。结果:4次免疫后7 d,小鼠鼻相关淋巴组织(NALT)、鼻通道(NP)和Peyer’s结(PP)淋巴细胞中,CD3+T细胞数均显著增加,其中以CD4+T细胞增加为主。FSM-2117免疫组的脾细胞中B220+细胞显著增加;而FS+5416免疫组的脾细胞中CD3+T细胞显著增加。4次免疫后30 d,NALT、NP和脾淋巴细胞仍出现上述变化;而90 d,仅NALT和NP淋巴细胞出现上述同样变化。结论:两株双价痢疾菌苗滴鼻免疫小鼠后,能有效地诱导黏膜和系统免疫应答,且持续时间较长,但该免疫应答的减弱是从距免疫部位较远的部位而开始的。

AIM: To observe the changes of lymphocyte phenotype in different lymphoid tissues of mice at various time after in-tranasal immunization with bivalent Shigella vaccines. METHODS: BALB/c mice were randomly divided into three groups, 30 mice per group. Mice were intranasally immunized respectively with PBS, FSM-2117 or FS-5416 four times (bacterial number was sequentially 5 × 106, 1 × 107, 4×107 and 4 × 107CFU/mouse) with two-week intervals. Lymphocytes in nasal associated lymphoid tissue (NALT), nasal passage (NP), spleen or Peyer's patch (PP) were isolated on the 7th, 30th and 90th day after the last immunization. The phenotype of the lymphocytes was detected by FACS. RESULTS: CDS+ T cells in NALT, NP and PP increased significantly on the 7th day after the immunization, in which most were CD4 + T cells. B220+ cells and CD3+ T cells increased notably in spleens of FSM-2117 group and FS-5416 group, respectively. The same phenotypic changes still maintained in the NALT, NP and spleen on the 30th day after immunization, but only present in NALT and NP on the 90th day. CONCLUSION: Intranasal inoculation with the two bivalent Shigella vaccines can effectively induce immune responses in different mucosal sites and systematically which can durate for a long time and start to weaken from the tissues distal from nose to those proximal.