The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surfa...The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surface antigen protein; when this mutated gene was expressed in monkey kidney cell line COS-M6, the protein product (S301 protein) could be secreted from the cells. Moreover, the inhibition of the secretion of the major surface antigen protein by this altered large surface antigen protein was greatly reduced, suggesting that the deleted region contained a retention sequence which prevented the secretion of the large surface antigen. However, the coexpression of the major S protein was essential for the secretion of the S301 protein. When coexpressed, the secretion of these two proteins was synchronous. Like the wild type large surface antigen protein, the S301 protein could be translocated into ertdoplasmic reticulum and glycosylated after its synthesis in COS cells. The S301 protein was thermostable and proteinase-resistant. It also retained the antigenicity of the large S and major S proteins. Given the fact that the S301 protein is readily secretable, stable and identical to the large S protein in terms of their antigenicity, it may be developed into a new generation of recombinant vaccine for the prevention of viral hepatitis.展开更多
目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方...目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方法:对83例HBeAg阴性的慢性乙型肝炎患者血清,采用ELISA进行LHBs检测,全自动免疫分析仪检测HBeAg,实时荧光定量PCR方法检测HBV DNA。分析HBV DNA无拷贝数组、低拷贝数组及高拷贝数组中LHBs的检出率以及HBV DNA不同拷贝数组与LHBs的相关性。结果:在HBV DNA无拷贝数组,LHBs的阳性率为6.38%;低拷贝数组,LHBs的阳性率为56.25%,吸光度值与拷贝数对数值没有显著的相关性(r=0.25,P=0.36);高拷贝数组,阳性率为95%,吸光度值与拷贝数对数值有良好相关性(r=0.90,P<0.0001)。结论:检测HBeAg阴性慢性乙型肝炎患者血清LHBs,有助于判断患者体内HBV的复制程度,但是否能够作为HBeAg阴性乙型肝炎抗病毒治疗终点的有效判定指标,尚有待进一步探讨。展开更多
基金This research was supported in part by U. S. Public Health Service Grants CA-70175 and CA-2243 to J. Mertz from the U. S. National Institute of Health.
文摘The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surface antigen protein; when this mutated gene was expressed in monkey kidney cell line COS-M6, the protein product (S301 protein) could be secreted from the cells. Moreover, the inhibition of the secretion of the major surface antigen protein by this altered large surface antigen protein was greatly reduced, suggesting that the deleted region contained a retention sequence which prevented the secretion of the large surface antigen. However, the coexpression of the major S protein was essential for the secretion of the S301 protein. When coexpressed, the secretion of these two proteins was synchronous. Like the wild type large surface antigen protein, the S301 protein could be translocated into ertdoplasmic reticulum and glycosylated after its synthesis in COS cells. The S301 protein was thermostable and proteinase-resistant. It also retained the antigenicity of the large S and major S proteins. Given the fact that the S301 protein is readily secretable, stable and identical to the large S protein in terms of their antigenicity, it may be developed into a new generation of recombinant vaccine for the prevention of viral hepatitis.
文摘目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方法:对83例HBeAg阴性的慢性乙型肝炎患者血清,采用ELISA进行LHBs检测,全自动免疫分析仪检测HBeAg,实时荧光定量PCR方法检测HBV DNA。分析HBV DNA无拷贝数组、低拷贝数组及高拷贝数组中LHBs的检出率以及HBV DNA不同拷贝数组与LHBs的相关性。结果:在HBV DNA无拷贝数组,LHBs的阳性率为6.38%;低拷贝数组,LHBs的阳性率为56.25%,吸光度值与拷贝数对数值没有显著的相关性(r=0.25,P=0.36);高拷贝数组,阳性率为95%,吸光度值与拷贝数对数值有良好相关性(r=0.90,P<0.0001)。结论:检测HBeAg阴性慢性乙型肝炎患者血清LHBs,有助于判断患者体内HBV的复制程度,但是否能够作为HBeAg阴性乙型肝炎抗病毒治疗终点的有效判定指标,尚有待进一步探讨。
