日本血吸虫重组Bb疫苗不同途径免疫BALB/c小鼠不同时间诱导的免疫应答作用

doi:10.13481/j.1671-587x.20150114

摘要/Abstract

摘要：

目的: 探讨日本血吸虫重组两歧双歧杆菌(rBb)疫苗不同途径免疫BALB/c小鼠后不同时间诱导的免疫应答作用,阐明该疫苗的免疫应答机制。方法: 将88只BALB/c小鼠随机分为2组,每组44只,分别经皮下注射(皮下注射组,SC组)和鼻腔黏膜(鼻腔黏膜接种组,IN组)接种免疫小鼠,在免疫后0~20周每2周每组随机取4只小鼠,无菌取脾,制备悬浮脾细胞,采用四甲基偶氮唑盐比色法(MTT)检测脾细胞增殖活性,流式细胞仪(FACsort)检测T细胞亚群和脾细胞凋亡率,双抗体夹心ELISA法检测脾细胞上清液中干扰素γ(IFN-γ)、白细胞介素10(IL-10)和白细胞介素17(IL-17)的动态变化。结果: 与免疫0周时比较,SC组小鼠脾细胞增殖活性于免疫后8周达到峰值(P<0.05),IN组小鼠于免疫后4周达到峰值(P<0.05)。与免疫0周时比较,SC组和IN组小鼠CD4⁺T细胞均于免疫后8周达到峰值(P<0.05);CD8⁺T细胞分别于8周和6周达到峰值,但差异无统计学意义(P>0.05)。与免疫0周时比较,2组小鼠脾细胞上清液中IFN-γ水平均于免疫后2周达到峰值(P<0.05),SC组和IN组IL-17水平分别于免疫后14和12周达到峰值(P<0.05)。与免疫0周时比较,SC组小鼠脾细胞凋亡率于免疫后2周达到峰值(P<0.05),IN组于4周达到峰值(P<0.05)。IN组与SC组小鼠各检测指标比较差异均无统计学意义(P>0.05)。结论: 经皮下注射和鼻腔黏膜途径接种的日本血吸虫重组Bb疫苗早期即可诱导小鼠产生有效的免疫应答,以Th1型免疫应答为主,2种免疫途径所诱导的免疫应答无明显差异。

关键词: 日本血吸虫, 重组两歧双歧杆菌(pGEX-Sj26GST)疫苗, 脾细胞, 细胞增殖, 细胞凋亡, T细胞亚群, 细胞因子

Abstract:

Objective To observe the immune responses of BALB/c mice after immunized by recombinant vaccine Bifidobacterium bifidum of Schistosoma japonicum in different ways for different time, and to explore the immune mechanism of the vaccines. Methods 88 BALB/c mice were randomly divided into two groups, 44 mice in each group, and they were subcutaneously (SC group) and intranasally(IN group) immunized, respectively.Four mice selected randomly from each group were sacrificed every 2 week during 0-20 weeks after immunization, and the splenocytes of them were isolated under sterile condition.The proliferation ability of splenocytes was investigated by MTT colorimetric assay;the subsets of CD4⁺ and CD8⁺ T cells and apoptotic rates of splenocyts were detected by FACsort flow cytometry, and the IFN-γ, IL-10 and IL-17 expression levels in supernatant of splenocytes were determined by ELISA method. Results Compared with 0 week, the proliferation acitivity of the splenocytes in SC group peaked at the 8th week;and reached the maximum at the 4th week after immunization in IN group(P<0.05).The CD4⁺ subsets reached the peak at the 8th week(P<0.05) in both groups;while the CD8⁺ subsets reached the maximum at the 8th week in SC group and at the 6th week in IN group respectively, but there was no significant difference (P> 0.05).Compared with 0 week, the apoptotic rates of splenocytes peaked at the 2th week in SC group and at the 4th week in IN group(P<0.05).Compared with 0 week, the level of IFN-γ in supernatant of splenocytes peaked at the 2th week in both groups(P<0.05).The levels of IL-17 in SC group and IN group reached the peak at the 14th and 12th week respectively( P<0.05).Between IN group and SC group, all the indexes above had no statistically significant differences(P>0.05). Conclusion The rBb (pGEX-Sj26GST) vaccine may play a protective immune response in both ways, which mainly induce Th1 immune response, and the responses have no significant differences between IN and SC groups.

Key words: Schistosoma japonicum, rBb(pGEX-Sj26GST)vaccine, splenocytes, cell proliferation, apoptosis, T cell subsets, cytokines

中图分类号:

R383.24

张丽, 李文桂, 谭建蓉. 日本血吸虫重组Bb疫苗不同途径免疫BALB/c小鼠不同时间诱导的免疫应答作用[J]. 吉林大学学报(医学版), 2015, 41(01): 69-76.

ZHANG Li, LI Wengui, TAN Jianrong. Immune responses in BALB/c mice immunized with recombinant vaccine Bifidobacterium bifidum of Schistosoma japonicum in different ways for different time[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 69-76.

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