表达PCV2 Cap蛋白重组复制缺陷型腺病毒黏膜免疫效果  

Evaluation of Mucosal Immunity of Replication Defective Recombinant Adenovirus Expressing Capsid Protein of PCV2

作　　者：邓祖丽颖 刘雨丰[2] 马宁宁[2] 陈陆 DENG Zuliying;LIU Yufeng;MA Ningning;CHEN Lu(Zhengzhou Preschool Education College,Zhengzhou 450000,China;Henan Agricultural University,Zhengzhou 450000,China)

机构地区：[1]郑州幼儿师范高等专科学校,河南郑州450000 [2]河南农业大学,河南郑州450000

出　　处：《河南农业科学》2020年第10期143-148,共6页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(31772781);河南省高校科技创新人才支持计划项目(14HASTIT022)。

摘　　要：为研发高效的猪圆环病毒2型(PCV2)黏膜疫苗,通过口服、肌内和滴鼻途径,将表达PCV2 Cap蛋白主要抗原表位的重组复制缺陷型腺病毒(rAd/Cap/518)免疫Balb/c小鼠,同时以无外源基因的空腺病毒(wild-rAd)经滴鼻途径免疫Balb/c小鼠作为对照组,分别检测小鼠血清中IgG和唾液、肺部、肠道灌洗液中IgA抗体水平,脾脏T淋巴细胞亚群及相关细胞因子IFN-γ和IL-4的分泌水平以及淋巴结、脾脏和肺组织中PCV2病毒载量,评价其黏膜免疫效果。结果显示,与对照组相比,rAd/Cap/518经肌内途径免疫诱导产生的血清IgG抗体水平显著增加;经滴鼻和口服途径免疫诱导产生的局部黏膜部位IgA抗体水平显著增加。rAd/Cap/518经滴鼻、肌内和口服3种途径免疫诱导产生的CD3+T细胞百分率为46.60%~55.65%,经滴鼻免疫诱导产生的CD3+CD4+T细胞百分率为34.43%~37.29%,经肌内和口服途径免疫诱导产生的CD3+CD4+T细胞百分率为36.35%~41.58%,经滴鼻途径免疫诱导的CD3+CD8+T细胞百分率为12.39%~18.32%,经口服免疫诱导的CD3+CD8+T细胞百分率为16.29%,均显著高于对照组。rAd/Cap/518经滴鼻途径免疫后,能诱导小鼠脾脏和肠系膜淋巴细胞分泌IFN-γ。攻毒后免疫鼠体内病毒载量显著降低。综上,rAd/Cap/518经肌内途径免疫主要诱导全身性免疫应答,经滴鼻和口服途径免疫能诱导全身性和局部黏膜免疫应答,是一种很有前景的黏膜免疫候选疫苗。To develop efficient porcine circovirus type 2(PCV2)vaccine,the mucosal immune effect of PCV2 Cap protein epitope recombinant replication-deficient adenovirus(rAd/Cap/518)was evaluated.The Balb/c mice were immunized by oral,intramuscular and intranasal routes with rAd/Cap/518,and the mice were immunized by intranasal routes with wild rads without exogenous genes as the control group.IgG in serum,IgA antibody in saliva,lung and intestinal lavage fluid,secretion of T lymphocyte subsets and related cytokines IFN-γand IL-4 in spleen,and PCV2 viral load in lymph nodes,spleen and lung were detected.The results showed that,compared with the control group,the level of serum IgG antibody induced by rAd/Cap/518 via intramuscular route increased significantly.The level of IgA antibody in local mucosa significantly increased by intranasal and oral immunization.The percentage of CD3+T cells induced by rAd/Cap/518 was 46.60%—55.65%by intranasal,intramuscular and oral routes.The percentage of CD3+CD4+T cells was 34.43%—37.29%,the percentage of CD3+CD4+T cells induced by intramuscular and oral immunization was 36.35%—41.58%,the percentage of CD3+CD8+T cells induced by intranasal immunization was 12.39%—18.32%,and the percentage of CD8+T cells induced by oral immunization was 16.29%,which were significantly higher than that of the control group.Intranasal immunization with rAd/Cap/518 could induce IFN-γproduction in spleen and mesenteric lymphocytes.The virus load of immunized mice decreased significantly.These results indicate that rAd/Cap/518 mainly induces systemic immune response via intramuscular immunization,and systemic and local mucosal immune response via intranasal and oral immunization.It is a promising mucosal vaccine candidate for PCV2.

关 键 词：猪圆环病毒2型 CAP蛋白 复制缺陷型腺病毒 黏膜免疫 免疫效果 

分 类 号：S852.65[农业科学—基础兽医学]