AIM: To explore how to improve the immunogenicity of HBcAg CTL epitope based polypeptides and to trigger an HBV-specific HLA I-restricted CD8^+ T cell response in vitro.METHODS: A new panel of mimetic therapeutic pept...AIM: To explore how to improve the immunogenicity of HBcAg CTL epitope based polypeptides and to trigger an HBV-specific HLA I-restricted CD8^+ T cell response in vitro.METHODS: A new panel of mimetic therapeutic peptides based on the immunodominant B cell epitope of HBV PreS2 18-24 region, the CTL epitope of HBcAg18-27 and the universal T helper epitope of tetanus toxoid (TT) 830-843 was designed using computerized molecular design method and synthesized by Merrifield's solid-phase peptide synthesis.Their immunological properties of stimulating activation and proliferation of lymphocytes, of inducing TN1 polarization,CD8^+ T cell magnification and HBV-specific CD8^+ CTL mediated cytotoxicity were investigated in vitro using HLA-A2^+ human peripheral blood mononuclear cells (PBMCs) from healthy donors and chronic hepatitis B patients.RESULTS: Results demonstrated that the therapeutic polypeptides based on immunodominant HBcAg18-27 CTL,PreS2 B- and universal TN epitopes could stimulate the activation and proliferation of lymphocytes, induce specifically and effectively CD8+ T cell expansion and vigorous HBVspecific CTL-mediated cytotoxicity in human PBMCs.CONCLUSION: It indicated that the introduction of immunodominant T helper plus B-epitopes with short and flexible linkers could dramatically improve the immunogenicity of short CTL epitopes in vitro.展开更多
目的检测来源于HCA587的HLA-A2限制性表位肽免疫HLA-A2转基因小鼠后产生细胞应答的水平,筛选出体内具有免疫原性的HLA-A2限制性表位肽。方法将来源于HCA587的候选HLA-A2限制性肽与MHC-Ⅱ类限制性肽HBV-core128-140、不完全弗氏佐剂、Cp ...目的检测来源于HCA587的HLA-A2限制性表位肽免疫HLA-A2转基因小鼠后产生细胞应答的水平,筛选出体内具有免疫原性的HLA-A2限制性表位肽。方法将来源于HCA587的候选HLA-A2限制性肽与MHC-Ⅱ类限制性肽HBV-core128-140、不完全弗氏佐剂、Cp G ODN1826联合应用皮下免疫HLA-A2转基因小鼠,用酶联免疫斑点实验(ELISpot)检测小鼠脾细胞表位肽特异性细胞免疫应答的水平,流式细胞术检测表位肽特异性的CD8+T细胞比例。结果在检测的4个表位肽中,p248-256诱导HLA-A2转基因小鼠产生的细胞免疫应答最强,HLA-A2-H-2Kb-p248-256四聚体染色进一步表明p248-256免疫后可产生p248-256特异性的CD8+T细胞。结论来源于HCA587的HLA-A2限制性表位肽中,p248-256具有较强的体内免疫原性。展开更多
目的通过检测湖北土家族人群HLA-A2超型(包含A*0201、A*0202、A*0203、A*0204、A*0205、A*0206、A*0207、A*6802、A*6901)各等位基因,分析与宫颈癌关联的等位基因及其结构与功能特点.方法提取湖北土家族正常人群236名育龄妇女及59例原...目的通过检测湖北土家族人群HLA-A2超型(包含A*0201、A*0202、A*0203、A*0204、A*0205、A*0206、A*0207、A*6802、A*6901)各等位基因,分析与宫颈癌关联的等位基因及其结构与功能特点.方法提取湖北土家族正常人群236名育龄妇女及59例原发性宫颈癌患者外周血DNA,采用SBT(sequence based typing)、SSOP(Sequence Specific Oligonucleotide Probes)HLA基因分型技术,对HLA-A2超型各等位基因型进行分型,比较宫颈癌病例组和正常对照组中HLA-A2超型中相应等位基因型构成比的差异,并分析相关等位基因的结构特点.结果有7种HLA-A2超型等位基因HLA-A*0201(17.3%)HLA-A*0202(9.5%)HLA A*0203(1.4%)HLA-A*0204(3.8%)HLA-A*0205(3.1%)HLA-A*0206(10.8%)HLA-A*0207/0215N(9.8%),其中HLA-A*0202和HLA A*0206在正常对照组和宫颈癌病人组的构成比有显著性差异(p＜0.05),HLA-A*0202(OR=0.24,95%CI=0.05～0.48)和HLA A*0206(OR=0.2,95%CI=0.67～1.07)对于宫颈癌发生的易感性有保护作用.结论湖北土家族人群HLA-A*0201所占比例最高达17.3%,HLA-A*0202和HLA-A*0206对于宫颈癌发生的易感性有保护作用.展开更多
基金Supported by the National Natural Science Foundation of China,No.30271189the National 973 Project,No.2001CB510001
文摘AIM: To explore how to improve the immunogenicity of HBcAg CTL epitope based polypeptides and to trigger an HBV-specific HLA I-restricted CD8^+ T cell response in vitro.METHODS: A new panel of mimetic therapeutic peptides based on the immunodominant B cell epitope of HBV PreS2 18-24 region, the CTL epitope of HBcAg18-27 and the universal T helper epitope of tetanus toxoid (TT) 830-843 was designed using computerized molecular design method and synthesized by Merrifield's solid-phase peptide synthesis.Their immunological properties of stimulating activation and proliferation of lymphocytes, of inducing TN1 polarization,CD8^+ T cell magnification and HBV-specific CD8^+ CTL mediated cytotoxicity were investigated in vitro using HLA-A2^+ human peripheral blood mononuclear cells (PBMCs) from healthy donors and chronic hepatitis B patients.RESULTS: Results demonstrated that the therapeutic polypeptides based on immunodominant HBcAg18-27 CTL,PreS2 B- and universal TN epitopes could stimulate the activation and proliferation of lymphocytes, induce specifically and effectively CD8+ T cell expansion and vigorous HBVspecific CTL-mediated cytotoxicity in human PBMCs.CONCLUSION: It indicated that the introduction of immunodominant T helper plus B-epitopes with short and flexible linkers could dramatically improve the immunogenicity of short CTL epitopes in vitro.
文摘目的探讨人类白细胞抗原(human leukocyte antigen,HLA)基因多态性与乙型肝炎病毒(HBV)感染的相关性。方法收集云南省昆明市延安医院健康体检者静脉血样本501例,采用酶联免疫吸附试验(ELISA)检测HBV二对半,根据HBV二对半检测结果分为HBV携带组和既往感染组以及健康对照组3组,用序列特异性引物聚合酶链反应(polymerase chain reaction with sequence specific primers,PCR-SSP)基因分型技术检测HLA-A抗原的基因型,将HBV携带组和健康对照组以及HBV既往感染组和健康对照组的HLA-A基因多态性的分布频率进行比较。采用SPSS17.0软件进行数据统计分析。结果健康对照组HLA-A2阳性数占比47.49%,等位基因频率数占比31.29%;健康对照组基因分布频率总体与中华骨髓库发布的中国常见及确认的HLA-A等位基因表一致。HBV携带组HLA-A2阳性数占比63.04%,等位基因频率数占比42.23%,携带者的HLA-A2阳性率和等位基因频率差异有统计学意义(P<0.05);HBV既往感染组HLA-A2阳性数占比56.14%,等位基因频率数占比35.97%,既往感染组的HLA-A2阳性率和等位基因频率差异无统计学意义(P>0.05)。结论HLA-A2基因可能是慢性乙型肝炎HBV携带者的易感基因。
文摘目的检测来源于HCA587的HLA-A2限制性表位肽免疫HLA-A2转基因小鼠后产生细胞应答的水平,筛选出体内具有免疫原性的HLA-A2限制性表位肽。方法将来源于HCA587的候选HLA-A2限制性肽与MHC-Ⅱ类限制性肽HBV-core128-140、不完全弗氏佐剂、Cp G ODN1826联合应用皮下免疫HLA-A2转基因小鼠,用酶联免疫斑点实验(ELISpot)检测小鼠脾细胞表位肽特异性细胞免疫应答的水平,流式细胞术检测表位肽特异性的CD8+T细胞比例。结果在检测的4个表位肽中,p248-256诱导HLA-A2转基因小鼠产生的细胞免疫应答最强,HLA-A2-H-2Kb-p248-256四聚体染色进一步表明p248-256免疫后可产生p248-256特异性的CD8+T细胞。结论来源于HCA587的HLA-A2限制性表位肽中,p248-256具有较强的体内免疫原性。
文摘目的通过检测湖北土家族人群HLA-A2超型(包含A*0201、A*0202、A*0203、A*0204、A*0205、A*0206、A*0207、A*6802、A*6901)各等位基因,分析与宫颈癌关联的等位基因及其结构与功能特点.方法提取湖北土家族正常人群236名育龄妇女及59例原发性宫颈癌患者外周血DNA,采用SBT(sequence based typing)、SSOP(Sequence Specific Oligonucleotide Probes)HLA基因分型技术,对HLA-A2超型各等位基因型进行分型,比较宫颈癌病例组和正常对照组中HLA-A2超型中相应等位基因型构成比的差异,并分析相关等位基因的结构特点.结果有7种HLA-A2超型等位基因HLA-A*0201(17.3%)HLA-A*0202(9.5%)HLA A*0203(1.4%)HLA-A*0204(3.8%)HLA-A*0205(3.1%)HLA-A*0206(10.8%)HLA-A*0207/0215N(9.8%),其中HLA-A*0202和HLA A*0206在正常对照组和宫颈癌病人组的构成比有显著性差异(p＜0.05),HLA-A*0202(OR=0.24,95%CI=0.05～0.48)和HLA A*0206(OR=0.2,95%CI=0.67～1.07)对于宫颈癌发生的易感性有保护作用.结论湖北土家族人群HLA-A*0201所占比例最高达17.3%,HLA-A*0202和HLA-A*0206对于宫颈癌发生的易感性有保护作用.
