河南农业科学 ›› 2020, Vol. 49 ›› Issue (10): 143-148.DOI: 10.15933/j.cnki.1004-3268.2020.10.020

所属专题: 动物新型疫苗专题

• 畜牧·兽医 • 上一篇    下一篇

表达PCV2 Cap蛋白重组复制缺陷型腺病毒黏膜免疫效果

邓祖丽颖1,2,刘雨丰2,马宁宁2,陈陆2   

  1. (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)
  • 收稿日期:2020-04-12 出版日期:2020-10-15 发布日期:2020-10-15
  • 通讯作者: 陈陆( 1971- ),男,河南罗山人,教授,博士,主要从事病毒分子生物学研究。E-mail:chenluhau@126.com
  • 作者简介:邓祖丽颖(1969-),女,河南郑州人,副教授,硕士, 主要从事动物生物技术研究。E-mail:892743592@qq.com
  • 基金资助:
    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)

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

DENG Zuliying1,2,LIU Yufeng2,MA Ningning2,CHEN Lu2   

  1. (1.Zhengzhou Preschool Education College,Zhengzhou 450000,China;2.Henan Agricultural University,Zhengzhou 450000,China)
  • Received:2020-04-12 Published:2020-10-15 Online:2020-10-15

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。

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

Abstract: 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-γ and 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-γ production 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.

Key words: Porcine circovirus type 2, Cap protein, Replication defective recombinant adenovirus, Mucosal immunity, Immune efficiency

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