CD40L对单纯疱疹病毒2型DNA疫苗的免疫增强作用研究

发布时间：2018-05-07 01:28

本文选题：CD配体(CDL) + 单纯疱疹病毒型(HSV-)　；参考：《医学研究杂志》2015年10期

【摘要】：目的研究重组质粒pc DNA3-Kan/CD40L辅佐HSV-2 DNA疫苗增强小鼠体液免疫和细胞免疫的作用效果,探讨其作为HSV-2 DNA疫苗佐剂的潜力。方法 (1)构建鼠CD40L基因的重组真核表达质粒pc DNA3-Kan/CD40L。(2)体外细胞实验:检测重组质粒p CD40L刺激小鼠外周血淋巴细胞增殖情况和脾细胞分泌IFN-γ的能力。(3)体内动物实验:48只雌性BALB/c小鼠随机分为4个免疫组p K组、pg D组、pc CD40L+pg D组和p K+pg D组。小鼠后腿肌内注射,共免疫2次,间隔3周。末次免疫3周后,每组随机取4只小鼠进行致死剂量攻毒实验验证疫苗的保护作用。ELISA检测小鼠血清抗HSV-2 Ig G抗体水平和趋化因子RANTES;流式细胞术检测全血中CD4+和CD8+T细胞百分率以及分泌IFN-γ和IL-4的T细胞的百分率;MTS法检测小鼠脾脏T细胞的增殖能力。结果 (1)体外实验结果:重组质粒pc DNA3-Kan/CD40L对小鼠外周血淋巴细胞的增殖能力和刺激脾细胞分泌IFN-γ的能力均显著大于空质粒pc DNA3-Kan(P0.05)。(2)体内实验结果:小鼠血清抗HSV-2 Ig G水平、趋化因子RANTES、脾淋巴细胞刺激指数和外周血CD4+T细胞数和分泌IFN-γ的Th1细胞数均高于其他免疫组(P0.05)。pc DNA3-Kan/CD40L+pg D组预防小鼠感染HSV-2效果好于其他免疫组。结论 (1)重组质粒pc DNA3-Kan/CD40L能够诱导外周血淋巴细胞增殖并刺激脾细胞分泌IFN-γ具有作为疫苗佐剂的潜力。(2)pc DNA3-Kan/CD40L可以辅助HSV-2 DNA疫苗诱导BALB/c小鼠产生特异性抗HSV-2的体液免疫和细胞免疫,具备作为HSV-2 DNA疫苗免疫佐剂的能力。
[Abstract]:Objective to study the effect of recombinant plasmid PC DNA3-Kan/CD40L and HSV-2 DNA vaccine on enhancing humoral and cellular immunity in mice, and to explore its potential as adjuvant of HSV-2 DNA vaccine. Methods 1) Construction of recombinant eukaryotic expression plasmid of mouse CD40L gene pcDNA3-Kanr / CD40L.2) in vitro cell experiment: the proliferation of peripheral blood lymphocytes stimulated by recombinant plasmid p CD40L and the ability of spleen cells to secrete IFN- 纬 were detected. Female BALB/c mice were randomly divided into four immunization groups: PK group, PG D group, PC CD40L PG D group and p K PG D group. Mice were injected intramuscularly into hind leg and immunized twice at intervals of 3 weeks. Three weeks after the last immunization, Four mice in each group were randomly selected for lethal dose attack test to verify the protective effect of the vaccine. Elisa was used to detect the level of anti HSV-2 IgG antibody and chemokine RANTES, flow cytometry was used to detect the percentage of CD4 and CD8 T cells in whole blood, and Percentage of T cells secreting IFN- 纬 and IL-4. Results 1) the results of in vitro experiment: the proliferation ability of recombinant plasmid PC DNA3-Kan/CD40L on peripheral blood lymphocytes of mice and the ability of stimulating spleen cells to secrete IFN- 纬 were significantly higher than those of empty plasmid PC DNA3-Kann P0.05G.) the results of in vivo experiment showed that the level of anti HSV-2 Ig G in serum of mice was significantly higher than that of blank plasmid PC DNA3-Kann P0.05G. The chemokine RANTES, spleen lymphocyte stimulating index, the number of CD4 T cells and the number of Th1 cells secreting IFN- 纬 in peripheral blood were better than those in other immune groups in preventing HSV-2 infection in mice. Conclusion the recombinant plasmid PC DNA3-Kan/CD40L can induce the proliferation of peripheral blood lymphocytes and stimulate the secretion of IFN- 纬 by splenocytes. It has the potential to be a vaccine adjuvant. The recombinant pc DNA3-Kan/CD40L can assist HSV-2 DNA vaccine to induce specific humoral and cellular immunity against HSV-2 in BALB/c mice. Ability to act as an immune adjuvant for HSV-2 DNA vaccines.
【作者单位】：温州医科大学检验医学院生命科学学院;浙江省医学科学院;
【基金】：浙江省自然科学基金资助项目(LY12H19009)
【分类号】：R392

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