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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 TGFβ1抗原表位插入乙型肝炎核心抗原(HBcAg)的免疫优势区c／el区, 原核表达蛋白作为重组TGF β1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。
Objective To investigate the anti-TGFβ1 neutralizing activity of anti-TGFβ1 induced by recombinant TGFβ1 vaccine in Balb / c mice and the anti-fibrosis effect. Methods The TGFβ1 epitope was inserted into the c / el region of the immunodominant region of HBcAg. The prokaryotic expression protein was used as a recombinant TGF β1 vaccine to induce high titer neutralizing antibody. ELISA was used to detect the immune response Serum antibody titers and mink lung epithelial cell growth inhibition assay were used to detect the in vitro neutralization of TGFβ1 by the antibody serum. The vaccine was used to prevent liver fibrosis in CTC model mice. Results Recombinant TGFβ1 vaccine induced high titers of anti-TGFβ1 in mice and combined with the expressed fusion protein and antigen peptide, blocking the inhibitory activity of TGFβ1 on mink lung epithelial cells and alleviating liver fibrosis in mice. Conclusion Recombinant TGFβ1 vaccine containing HBcAg as a carrier protein can induce high titers of neutralizing antibodies and block TGFβ1 activity in vitro and can be used as anti-fibrotic vaccine in animal models of anti-fibrosis.