| 曾铁兵,赵宇龙,唐一之,陈德军,何宇星,赵飞骏,徐青峰,万佳,尹浩泉.重组梅毒螺旋体黏附素在小鼠模型中的免疫原性及免疫保护性研究.[J].中南医学科学杂志.,2021,(3):265-269. |
| 重组梅毒螺旋体黏附素在小鼠模型中的免疫原性及免疫保护性研究 |
| Immunogenicity and immune protection of recombinant Treponema pallidum adhesins in murine model |
| 投稿时间：2021-02-24 修订日期：2021-03-15 |
| DOI：10.15972/j.cnki.43-1509/r.2021.03.004 |
| 中文关键词: 梅毒螺旋体 黏附素 免疫原性 免疫保护性 小鼠模型 |
| 英文关键词:Treponema pallidum adhesin immunogenicity immune protection mouse model |
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| 中文摘要: |
| 目的观察重组梅毒螺旋体(Tp)黏附素Tp0751(rTp0751)、Tp0136氨基端(rTp0136N)、Tp0435(rTp0435)诱生C57BL/6小鼠适应性免疫应答水平及免疫后抑制Tp在小鼠体内播散情况,为筛选梅毒疫苗候选分子提供依据。方法表达和纯化重组蛋白rTp0751、rTp0136N和rTp0435。将C57BL/6小鼠随机分为PBS对照组、rTp0751组、rTp0136N组、rTp0435组,分别以PBS、rTp0751、rTp0136N及rTp0435免疫各组小鼠3次；ELISA检测各组小鼠免疫血清特异性IgG抗体水平；流式细胞术(FCM)检测末次免疫后2周小鼠脾细胞胞内白细胞介素-4(IL-4)与干扰素-γ(IFN-γ)含量；实时荧光定量PCR(qPCR)检测Tp攻击6周后小鼠心、脾、脑组织中Tp的DNA载量。结果各免疫组小鼠血清特异性IgG抗体随免疫时间增加而逐渐升高并于首次免疫第6周达到峰值。FCM检测结果显示各免疫组淋巴细胞胞内CD4＋IFN-γ＋与CD8＋ IFN-γ＋含量均高于PBS对照组(P＜0.01),CD4＋T细胞产生IL-4水平均极低。qPCR检测结果显示,在脾脏、心脏和脑组织中,rTp0751与rTp0136N组Tp-DNA载量均低于PBS对照组(P＜0.05),rTp0435组与PBS对照组之间无明显差异。结论rTp0751、rTp0136N、rTp0435均具有良好的免疫原性,Tp0751、Tp0136N有望为梅毒候选疫苗分子。 |
| 英文摘要: |
| To observe the adaptive immune response of C57BL/6 mice induced by recombinant Treponema pallidum (Tp) adhesins Tp0751 (rTp0751), the amino terminal of Tp0136 (rTp0136N) and Tp0435 (rTp0435), and to provide the basis for screening the candidate molecules of syphilis vaccine. MethodsThe recombinant proteins Tp0751, Tp0136N and Tp0435 were expressed and purified. C57BL/6 mice were randomly divided into PBS control group, rTp0751 group, rTp0136N group and rTp0435 group. The mice were immunized with PBS, purified rTp0751, rTp0136N and rTp0435 for three times, respectively. The levels of serum specific IgG against recombinant antigens were detected by ELISA. Flow cytometry (FCM) was performed to detect the levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in spleen cells of mice 2 weeks after the last immunization. Quantitative real-time fluorescence PCR (qPCR) was used to detect the Tp-DNA load in the heart, spleen and brain of mice at the 6th week post Tp challenge. ResultsThe levels of serum specific IgG increased gradually and reached the peak at the 6th week after the first immunization. CD4＋IFN-γ＋ and CD8＋IFN-γ＋ in lymphocytes of mice immunized with the recombinant proteins were higher than those of the PBS control group (P＜0.01), with very low level of IL-4 produced by CD4＋ T cells in all mice. In spleen, heart and brain, the load of Tp-DNA in rTp0751 and rTp0136N immunization groups was lower than that in PBS group (P＜0.05), with no significant difference between rTp0435 immunization group and PBS group. ConclusionrTp0751, rTp0136N and rTp0435 show good immunogenicity in mouse model. Tp0751 and Tp0136N may be promising candidate molecules for syphilis vaccine. |
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