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机构地区:[1]中国医学科学院北京协和医学院医学生物学研究所,昆明650118
出 处:《中国医药生物技术》2015年第4期346-350,共5页Chinese Medicinal Biotechnology
基 金:云南省应用基础研究面上项目(2010CD135);教育部留学回国人员启动资金(北京协和医学院留学回国人员启动资金)
摘 要:目的确定并评价大肠杆菌中重组表达的轮状病毒肠毒素NSP4肽段aa112-175的免疫佐剂活性,并与商品化的铝佐剂进行比较。方法以葛兰素史克公司生产的2011-2012季节性流感病毒裂解疫苗福禄立适(Fluarix)为抗原,通过小鼠平行免疫实验,检测经纯化后冻干的重组肽段aa112-175、铝佐剂的佐剂活性。得到了初次免疫后3周,小鼠Fluarix特异的Ig G水平,以及加强免疫后2周,小鼠Fluarix特异的Ig G水平和IFN-γ水平数据,进而比较两种佐剂的活性,判定重组轮状病毒NSP4肽段aa112-175的佐剂活性水平。结果轮状病毒肠毒素NSP4肽段aa112-175在大肠杆菌中获得分泌表达,初次免疫和加强免疫后,NSP4 aa112-175佐剂的血清特异Ig G水平与铝佐剂组相当,特异的IFN-γ水平约为铝佐剂组的2倍。结论轮状病毒肠毒素NSP4重组肽段aa112-175经肌肉注射免疫小鼠后,其加强流感疫苗的体液免疫反应的佐剂活性与铝佐剂相似,细胞免疫反应强于铝佐剂。Objective To identify and evaluate the immune adjuvant activity of recombinant enterotoxin NSP4 peptide (aal 12-175) expressed in E.coli, and compare it with the aluminum adjuvant.Methods To determine the adjuvant activity of recombinant peptide (aa112-175), mice were vaccinated by Fluarix produced by GlaxoSmithKline 2011-2012 seasonal influenza virus antigen with recombinant rotavirus enterotoxin NSP4 peptide (aa112-175) or aluminum adjuvant (Inject alum-Thermo scientific, Number 77161). After the initial immunization 3 weeks and booster immunization 2 weeks later, the Fluarix specific IgG and IFN-γ levels were tested, respectively.Results The rotavirus enterotoxin NSP4 peptide (aa112-175) was secreting expression in E. coll. After the primary and booster immunizations, Fluarix specific IgG levels of recombinant peptide (aa112-175) group were equivalent to that of the aluminum adjuvant group. However, the IFN-γ level of the recombinant peptide (aa112-175) group is two times of that of the aluminum adjuvant group. Conelusion Adjuvant activity, especially Fluarix specific IgG levels, of recombinant rotavirus enterotoxin NSP4 peptide (aa112-175) are equal to that of aluminum adjuvant. However, IFN-γ level of the recombinant peptide (aa112-175) group is higher than that of the aluminum adjuvant.
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