重组转化生长因子β1疫苗诱导产生抗体的活性检测及抗纤维化作用

目的探讨重组转化生长因子（TGF）β1疫苗诱导Balb／c小鼠产生的抗体对TGFβ1体外中和活性检测及抗肝纤维化作用。方法TGFDl抗原表位插入乙型肝炎核心抗原（HBcAg）的免疫优势区c／e1区，原核表达蛋白作为重组TGFβ1疫苗免疫小鼠，诱导产生高效价中和抗体，酶联免疫吸附法检测免疫血清抗体滴度，貂肺上皮细胞生长抑制法检测抗体血清对TGFβ1体外中和活性，将疫苗用于四氯化碳造模的小鼠预防肝纤维化。结果重组TGFβ1疫苗可以诱导小鼠产生高滴度的抗-TGFβ1并与表达的融合蛋白及抗原肽结合，可以阻断TGFβ1对貂肺上皮细胞的抑制活性，减轻小鼠肝纤维化。结论HBCAg作为载体蛋白构建的重组TGFβ1疫苗可以诱导高滴度的中和抗体产生，在体外阻断TGFβ1活性，可作为抗纤维化疫苗用于抗纤维化动物模型。

Detecting the activity of antibodies induced by recombinant TGFβ1 vaccine

OBJECTIVE: To measure the neutralization activity in vitro of the antibodies induced by recombinant TGFbeta1 vaccine and to evaluate the vaccine's anti-liver fibrosis activity. METHODS: Balb/c mice were immunized with a fusion protein of the human TGFbeta1 epitope-inserted into a hepatitis B core antigen using a prokaryotic expression system. The antibody produced by the recombinant vaccine was determined using ELISA. The biological activity of the anti-TGFbeta1 antibody induced by the vaccine was measured by MTT using mink lung epithelial cell Mv-1-Lu as inhibiting cells. The fusion protein was used as a vaccine in a mice hepatic-fibrosis model. RESULTS: A high titer of anti-TGFbeta1 antibody and a low of anti-HBc antibody were detected in the mice after the immunization. The serum antibodies induced combined with the fusion and antigenic peptide prevented the TGFbeta1 inhibiting activity in the Mv-1-Lu cell. CONCLUSION: Recombinant fusion protein can be used as a cytokine vaccine to induce high titers of anti-TGFbeta1 antibodies. Our results show the potentiality of the fusion protein to be used as a vaccine for preventing liver fibrosis.