目的探讨慢性乙型肝炎患者外周血单个核细胞(PBMC)中转录因子ROG、GATA3和T-bet m RNA水平变化及意义。方法收集135例慢性乙型肝炎患者(轻度45例、中度42例、重度48例)和15例健康志愿者(正常对照组)的外周血,分离单个核细胞,提取总RNA,...目的探讨慢性乙型肝炎患者外周血单个核细胞(PBMC)中转录因子ROG、GATA3和T-bet m RNA水平变化及意义。方法收集135例慢性乙型肝炎患者(轻度45例、中度42例、重度48例)和15例健康志愿者(正常对照组)的外周血,分离单个核细胞,提取总RNA,用实时定量PCR的方法检测ROG、GATA3和T-bet m RNA水平。应用SPSS16.0软件进行统计分析。结果慢性乙型肝炎轻度、中度组和重度组患者外周血PBMCs中T-bet m RNA表达水平均明显高于健康对照组人群,组内比较差异均具有统计学意义(P<0.05)。慢性乙型肝炎重度组患者外周血PBMCs中ROG m RNA表达水平明显高于健康对照组人群、轻度组和中度组患者,差异均有统计学意义(P<0.05)。而轻度组、中度组患者和健康对照组比较差异均无统计学意义(P>0.05)。慢性乙型肝炎中度组和重度组患者外周血PBMCs中GATA3 m RNA表达水平明显高于健康对照组人群和轻度组患者,差异均具有统计学意义(P<0.05);而轻度组和健康对照、中度组患者和重度组比较差异均无统计学意义(P>0.05)。慢性乙型肝炎轻度、中度组和重度组患者T-bet/GATA3比值明显高于健康对照组人群,差异均具有统计学意义(P<0.05);但3组之间比较差异无统计学意义(P>0.05)。ROG表达水平与GATA3和T-bet/GATA3比值均无相关性(P>0.05)。结论 ROG、GATA3和T-bet在慢性乙型肝炎患者外周血PBMCs中表达水平上调,并参与疾病的发生和疾病进展。ROG在纠正和维持Th1/Th2的新平衡状态过程中起重要作用。展开更多
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
The northern pig-tailed macaque （Macaca leonina） has been identified as an independent species of Old World monkey, and we previously found that PBMCs from M. leonina were susceptible to human immunodeficiency virus...The northern pig-tailed macaque （Macaca leonina） has been identified as an independent species of Old World monkey, and we previously found that PBMCs from M. leonina were susceptible to human immunodeficiency virus type 1 （HIV-1）, which may be due to the absence of a TRIM5 protein restricting HIV-1 replication. Here we investigated the infection potentials of six laboratory adapted HIV-1 strains and three primary HIV-1 isolates in PBMCs from M. leonina. The results indicate that these strains are characterized by various but low replication levels, and among which, HIV-INL4-3 shows the highest replication ability. Based on the abundant evidence of species-specific interactions between restriction factors APOBEC3 and HIV/SIV-derived Vif protein, we subsequently examined the replication potentials of v/f-substituted HIV-1 （HSIV） in M. leonina PBMCs. Notably, HSIV-vifmac and stHIV-lsv chimeras, two HIV-1Ni.4-3-derived viruses encoding the viral infectivity factor （Vif） protein from SIVmac239, replicated robustly in cells from M. leonina, which suggests that HSIV could effectively antagonize the antiviral activity of APOBEC3 proteins expressed in cells of M. leonina. Therefore, our data demonstrate that M. leonina has the potential to be developed into a promising animal model for human AIDS.展开更多
文摘目的探讨慢性乙型肝炎患者外周血单个核细胞(PBMC)中转录因子ROG、GATA3和T-bet m RNA水平变化及意义。方法收集135例慢性乙型肝炎患者(轻度45例、中度42例、重度48例)和15例健康志愿者(正常对照组)的外周血,分离单个核细胞,提取总RNA,用实时定量PCR的方法检测ROG、GATA3和T-bet m RNA水平。应用SPSS16.0软件进行统计分析。结果慢性乙型肝炎轻度、中度组和重度组患者外周血PBMCs中T-bet m RNA表达水平均明显高于健康对照组人群,组内比较差异均具有统计学意义(P<0.05)。慢性乙型肝炎重度组患者外周血PBMCs中ROG m RNA表达水平明显高于健康对照组人群、轻度组和中度组患者,差异均有统计学意义(P<0.05)。而轻度组、中度组患者和健康对照组比较差异均无统计学意义(P>0.05)。慢性乙型肝炎中度组和重度组患者外周血PBMCs中GATA3 m RNA表达水平明显高于健康对照组人群和轻度组患者,差异均具有统计学意义(P<0.05);而轻度组和健康对照、中度组患者和重度组比较差异均无统计学意义(P>0.05)。慢性乙型肝炎轻度、中度组和重度组患者T-bet/GATA3比值明显高于健康对照组人群,差异均具有统计学意义(P<0.05);但3组之间比较差异无统计学意义(P>0.05)。ROG表达水平与GATA3和T-bet/GATA3比值均无相关性(P>0.05)。结论 ROG、GATA3和T-bet在慢性乙型肝炎患者外周血PBMCs中表达水平上调,并参与疾病的发生和疾病进展。ROG在纠正和维持Th1/Th2的新平衡状态过程中起重要作用。
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.
基金Foundation items： This work was supported by the National Basic Research Program （2012CBA01305） the National Natural Science Foundation of China （81172876, U0832601, 81273251 and U 1202228） the Knowledge Innovation Program of CAS （KSCX2-EW-R-13, Y206A- 71181）, and the Key Scientific and Technological Program of China （2012ZX10001-007, 2013ZX10001-002）. Acknowledgements： We thank Prof. Guang-Xia GAO （Institute of Biophysics, Chinese Academy of Sciences） for kindly providing HSIV proviral plasmids.We also thank Long-Bao LV, Gui LI and Dong- Ti HUANG of Kunming Primate Research Center for their assistance in obtaining blood samples from northem pig-tailed macaques （M. leonina） and Chinese rhesus macaques.
文摘The northern pig-tailed macaque （Macaca leonina） has been identified as an independent species of Old World monkey, and we previously found that PBMCs from M. leonina were susceptible to human immunodeficiency virus type 1 （HIV-1）, which may be due to the absence of a TRIM5 protein restricting HIV-1 replication. Here we investigated the infection potentials of six laboratory adapted HIV-1 strains and three primary HIV-1 isolates in PBMCs from M. leonina. The results indicate that these strains are characterized by various but low replication levels, and among which, HIV-INL4-3 shows the highest replication ability. Based on the abundant evidence of species-specific interactions between restriction factors APOBEC3 and HIV/SIV-derived Vif protein, we subsequently examined the replication potentials of v/f-substituted HIV-1 （HSIV） in M. leonina PBMCs. Notably, HSIV-vifmac and stHIV-lsv chimeras, two HIV-1Ni.4-3-derived viruses encoding the viral infectivity factor （Vif） protein from SIVmac239, replicated robustly in cells from M. leonina, which suggests that HSIV could effectively antagonize the antiviral activity of APOBEC3 proteins expressed in cells of M. leonina. Therefore, our data demonstrate that M. leonina has the potential to be developed into a promising animal model for human AIDS.
