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目的 研究白细胞介素 6(IL 6)在非复制型痘苗病毒中的表达及其对重组痘苗病毒免疫效果的影响。方法 利用非复制型痘苗病毒表达载体pNEOCK1 1 β75IL6和重组病毒RVJ1 2 3 ,通过两步重组构建能同时表达IL 6和乙型肝炎病毒 (HBV)HBsAg的非复制型重组痘苗病毒RVJ1 2 3ΔCK1 1 β75IL6。免疫动物并观察其免疫效果。 结果 Southernblot证实痘苗病毒C、K片段间基因缺失的同时伴有IL 6基因的插入 ,IL 6和HBsAg可同时表达。鼻腔吸入分别免疫BALB c小鼠和新西兰白兔 ,ELISPOT实验证实免疫后两周小鼠肺淋巴细胞的抗 HBsAgIgA、IgG抗体分泌细胞 (ASC)数比对照组 (RVJ1 2 3ΔCK1 1 β75)显著增加 ,并可在小鼠血液、肺浸出液以及新西兰白兔血液、肺浸出液、其他分泌液样品中检测到抗 HBsAg的特异性的IgA、IgG抗体。与对照组相比 ,IgA、IgG抗体阳转率及抗体滴度提高。结论 非复制型痘苗病毒载体中表达的IL 6可增强其诱导的免疫反应 ,为细胞因子IL 6作为有效的载体疫苗佐剂提供了实验基础
Objective To study the expression of interleukin 6 (IL 6) in non-replicating vaccinia virus and its effect on the immunogenicity of recombinant vaccinia virus. Methods The non-replicating vaccinia virus expression vector pNEOCK1 1 β75IL6 and the recombinant virus RVJ1 2 3 were constructed by two-step recombination to construct a recombinant non-replicating vaccinia virus RVJ1 2 3ΔCK1 1 β75IL6 that can express both IL-6 and hepatitis B virus (HBsAg) simultaneously . Immunize animals and observe their immune effects. Results Southern blot showed that the insertions of IL 6 gene were deleted while the C and K fragments of vaccinia virus were deleted. IL 6 and HBsAg could be expressed at the same time. BALBc mice and New Zealand white rabbits were immunized with nasal inhalation respectively. ELISPOT assay confirmed that the number of anti-HBsAg IgG and IgG antibody secreting cells (ASC) in lung lymphocytes increased significantly in two weeks after immunization compared with the control group (RVJ1 2 3ΔCK1 1 β75) Anti-HBsAg-specific IgA and IgG antibodies can be detected in mouse blood, lung leachate and New Zealand white rabbit blood, lung leachate and other secretion samples. Compared with the control group, IgA, IgG antibody positive rate and antibody titer increased. CONCLUSION: IL6 expressed in non-replicating vaccinia virus vectors enhances its induction of immune responses and provides an experimental basis for the cytokine IL6 to be an effective carrier vaccine adjuvant