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目的考察热休克蛋白HSP70对呼吸道合胞病毒(RSV)重组蛋白抗原G1F/M2免疫应答的调节作用。方法构建并表达了Javelin-G1F/M2重组蛋白,在G1F/M2的N端引入Javelin序列,通过该序列可以实现重组蛋白与HSP70蛋白高亲和力结合。以HSP70:Javelin-G1F/M2复合物等免疫Balb/c小鼠,用乳酸脱氢酶释放法测定CTL活性,ELISA法测定IgG及其亚型IgG1和IgG2a抗体滴度,空斑抑制实验测定血清中和抗体。结果经表达和纯化获得了纯度达90%以上的融合蛋白;皮下免疫HSP70:Javelin-G1F/M2复合物可诱导高滴度的IgG抗体和强的CTL应答,其抗体滴度和CTL应答强度均优于Javelin-G1F/M2加铝佐剂腹腔免疫和单独Javelin-G1F/M2皮下免疫。HSP70:Javelin-G1F/M2复合物在诱导中和抗体方面与Javelin-G1F/M2加铝佐剂腹腔免疫基本相当。HSP70:Javelin-G1F/M2复合物可以诱导更强的IgG2a应答,IgG的亚型IgG1/IgG2a的比值明显低于Javelin-G1F/M2加铝佐剂腹腔免疫组和单独Javelin-G1F/M2皮下免疫组。结论 HSP70可能通过增强CTL应答进一步纠正Th2型优势应答,使得Th1/Th2应答更加平衡。
Objective To investigate the regulatory effect of heat shock protein HSP70 on the G1F / M2 immune response of respiratory syncytial virus (RSV) recombinant protein antigen. Methods The Javelin-G1F / M2 recombinant protein was constructed and expressed. The Javelin sequence was introduced into the N-terminus of G1F / M2. By this sequence, the recombinant protein could bind to HSP70 protein with high affinity. Balb / c mice were immunized with HSP70: Javelin-G1F / M2 complex, CTL activity was determined by lactate dehydrogenase release assay, IgG and IgG2a antibody titers of IgG and its subtypes were determined by ELISA, Neutralizing antibodies. Results The fusion protein with more than 90% purity was obtained through the expression and purification. The HSV-HSP70: Javelin-G1F / M2 complex could induce high titer of IgG antibody and strong CTL response with both antibody titer and CTL response intensity Superior to Javelin-G1F / M2 plus adjuvant intraperitoneal immunization and Javelin-G1F / M2 alone subcutaneous immunization. HSP70: The Javelin-G1F / M2 complex is essentially equivalent to intraperitoneal immunization of Javelin-G1F / M2 plus aluminum adjuvant in the induction of neutralizing antibodies. The HSP70: Javelin-G1F / M2 complex induced a stronger IgG2a response with a significantly lower ratio of IgG1 / IgG2a to Javelin-G1F / M2 plus adjuvant ip and Javelin-G1F / M2 alone group. Conclusion HSP70 may further correct Th2-type predominant response by enhancing CTL response, making the Th1 / Th2 response more balanced.