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目的获取重组人干扰素-λ1(hIFN-λ1)的原核表达产物并研究hIFN-λ1对兔角膜基质细胞的抗单纯疱疹病毒(HSV)作用。方法用含基因重组表达载体pGEX-4T-λ1的大肠杆菌BL21,经IPTG诱导表达和包涵体提取纯化后获取hIFN-λ1蛋白,采用微量滴定法测定其效价。采用组织块培养法培养兔眼角膜基质细胞,用微量细胞病变法比较hIFN-λ1、IFN-α标准品对兔眼角膜基质细胞的抗HSV作用。结果诱导含基因重组表达载体PGEX-4T-λ1的大肠杆菌BL21后,蛋白电泳显示出一条相对分子量约为33kd的蛋白条带;用IFN-α标准品校正其效价为62.5U/ml。hIFN-λ1抗HSV-I的作用与IFN-α相近,HSV-II的作用比IFN-α约弱50%。结论含基因重组表达载体pGEX-4T-λ1的大肠杆菌,经诱导表达和包涵体提取纯化后可获得hIFN-λ1,它对兔眼角膜基质细胞有一定的抗HSV作用。
Objective To obtain the prokaryotic expression product of recombinant human interferon-λ1 (hIFN-λ1) and study the anti-herpes simplex virus (HSV) effect of hIFN-λ1 on rabbit corneal stromal cells. Methods The recombinant plasmid pGEX-4T-λ1 was transformed into E.coli BL21 and induced by IPTG. The hIFN-λ1 protein was extracted and purified from inclusion bodies. The titer of hIFN-λ1 protein was determined by microtiter method. Rabbit corneal stromal cells were cultured by tissue culture method. The anti-HSV effects of hIFN-λ1 and IFN-α standards on corneal stromal cells of rabbits were compared by micro-cytopathic assay. Results After inducing E. coli BL21 containing the gene recombinant expression vector PGEX-4T-λ1, the protein electrophoresis showed a protein band with a relative molecular weight of about 33 kd. The titer was corrected to 62.5 U / ml with the IFN-α standard. The effect of hIFN-λ1 against HSV-I is similar to that of IFN-α, which is about 50% weaker than IFN-α. Conclusion The recombinant plasmid pGEX-4T-λ1 containing Escherichia coli can produce hIFN-λ1 after induced expression and inclusion body purification. It has certain anti-HSV effect on rabbit corneal stromal cells.