含CPE蛋白的新型口服CVB3黏膜疫苗对小鼠病毒性心肌炎的保护作用
首发时间:2015-11-27
摘要:目的 改良新型B3型柯萨奇病毒(CVB3)黏膜疫苗chitosan-pcDNA3.1-VP1,充分发挥壳聚糖作为口服疫苗载体的优势,增强其预防CVB3性心肌炎的效果。方法 通过化学偶联法将CPE蛋白偶联至chitosan后,经共凝聚法与pVP1质粒形成复合物,口服免疫小鼠,25μg质粒每次,共4次,末次免疫后2周检测CVB3特异性血清IgG、粪便IgA,脾脏和肠系膜淋巴结T细胞免疫应答和免疫保护效果。结果 (1)chitosan-CPE-pVP1显著提高了CVB3特异性血清IgG水平、粪便IgA水平。(2)该新型疫苗显著促进了脾脏和MLN部位特异性T细胞增殖和分泌IFN-γ的能力。(3)感染后心肌病理显示:与其它组相比,chitosan-CPE-pcDNA3.1-VP1组发病程度最轻,心肌基本正常。(4)免疫小鼠心肌细胞内病毒的免疫组化结果显示:chitosan-CPE-pcDNA3.1-VP1口服疫苗可有预防CVB3引起的病毒性心肌炎。结论 新型口服粘膜疫苗chitosan-CPE-pcDNA3.1-VP1可有效增强chitosan-pVP1疫苗预防病毒性心肌炎的效果。
关键词: CVB3 CPE蛋白 口服 黏膜疫苗 病毒性心肌炎
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A novel oral vaccine containing CPE protein prevents CVB3-induced myocarditis in mice
Abstract:Objective To exert the advantage of chitosan as an oral vaccine carrier and Enhance the effect of chitosan-pcDNA3.1-VP1 mucosal vaccine to prevent onset of coxsackievirus B3(CVB3)-induced myocarditis. Methods Chitosan-CPE which were coupled with chemical reagents was complexed with DNA by co-conjugation method. Then 25μg(DNA) of each complex was administered orally to male BALB/c mice 4 times biweekly.Two weeks after the final immunization,the levels of CVB3-specific serumIgG,fecalIgA, CVB3-specific cell immune response and immune protection were detected. Results chitosan-CPE-pcDNA3.1-VP1 vaccine orally immunization significantly increased the levels of CVB3-specific serum IgG and fecal IgA,and enhanced T cell proliferation and IFN-γ-secreting ability.HE staining 7days following 3x LD50 CVB3 infection showed that there were fewer inflammatory cells under the epicardium in the hearts of chitosan-CPE-pVP1 immunized mice compared with control mice.Immunohistochemistry analysis showed little CVB3 loading within hearts of chitosan-CPE-pVP1 immunized mice.Conclusion Novel chitosan-CPE-pVP1 oral vaccine could more effectively prevent CVB3-induced myocard- itis than chitosan-pVP1 vaccine.?????
Keywords: CVB3 CPE protein;oral administration;mucosal vaccine;CVB3
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含CPE蛋白的新型口服CVB3黏膜疫苗对小鼠病毒性心肌炎的保护作用
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