Hepatitis C virus(HCV) has a high propensity to establish chronic infections. Failure of HCV-infected individuals to activate effective antiviral immune responses is at least in part related to HCV-induced impairment ...Hepatitis C virus(HCV) has a high propensity to establish chronic infections. Failure of HCV-infected individuals to activate effective antiviral immune responses is at least in part related to HCV-induced impairment of dendritic cells(DCs) that play a central role in activating T cell responses. Although the impact of HCV on DC phenotype and function is likely to be more prominent in the liver, major HCV-induced alterations are detectable in peripheral blood DCs(pb DCs) that represent the most accessible source of DCs. These alterations include numerical reduction, impaired production of inflammatory cytokines and increased production of immunosuppressive IL10. These changes in DCs are relevant to our understanding the immune mechanisms underlying the propensity of HCV to establish persistent infection. Importantly, the noninvasive accessibility of pb DCs renders the analysis of these cells a convenient procedure that can be serially repeated in patient follow-up. Accordingly, the study of pb DCs in HCV-infected patients during conventional treatment with pegylated interferon and ribavirin indicated that restoration of normal plasmacytoid DC count may represent an additional mechanism contributing to the efficacy of the dual therapy. It also identified the pre-treatment levels of plasmacytoid DCs and IL10 as putative predictors of response to therapy. Treatment of chronic HCV infection is changing, as new generation direct-acting antiviral agents will soon be available for use in interferon-free therapeutic strategies. The phenotypic and functional analysis of pb DCs in this novel therapeutic setting will provide a valuable tool for investigating mechanisms underlying treatment efficacy and for identifying predictors of treatment response.展开更多
AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blo...AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blot.Quantitative real-time polymerase chain reaction(PCR)was used to detect intestinal integrin-αm RNA expression.Intestinal CD83,CD86,CD74,CD3 and AKT were detected by immunohistochemistry,Western blot and PCR.Phosphorylated-AKT(p-AKT)was detected by immunohistochemistry and Western blot.RESULTS:In the FHF group[D-galactosamine(D-Galn)+lipopolysaccharide(LPS)group],the mice began to die after 6 h;conversely,in the D-Galn and LPS groups,the activity of mice was poor,but there were no deaths.Immunohistochemistry results showed that in FHF,the expression of CD11b/c(7988400±385941vs 1102400±132273,P<0.05),CD83(13875000±467493 vs 9257600±400364,P<0.05),CD86(7988400±385941 vs 1102400±13227,P<0.05)and CD74(11056000±431427 vs 4633400±267903,P<0.05)was significantly increased compared with the normal saline(NS)group.Compared with the NS group,the protein expression of CD11b/c(5.4817±0.77 vs 1.4073±0.37,P<0.05)and CD86(4.2673±0.69 vs 1.1379±0.42,P<0.05)was significantly increased.Itg-α(1.1224±0.3 vs 0.4907±0.19,P<0.05),CD83(3.6986±0.40 vs 1.0762±0.22,P<0.05)and CD86(1.5801±0.32 vs 0.8846±0.10,P<0.05)m RNA expression was increased significantly in the FHF group.At the protein level,expression of CD74in the FHF group(2.3513±0.52)was significantly increased compared with the NS group(1.1298±0.33),whereas in the LPS group(2.3891±0.47),the level of CD74 was the highest(P<0.05).At the gene level,the relative expression of CD74 m RNA in the FHF group(1.5383±0.26)was also significantly increased in comparison to the NS group(0.7648±0.22;P<0.05).CD3 expression was the highest in the FHF group(P<0.05).In the FHF,LPS and D-Galn groups,the expression of AKT at the protein and m RNA levels was elevated compared with the NS group,but there wasno statistical significance(P>0.05).The p-AKT protein expr展开更多
文摘目的：探讨腺病毒载体介导HBV抗原基因修饰的树突状细胞(DCs)诱导抗HBV特异性CTL反应方法：制备携带HBsAg、HBeAg和HBcAg基因的3种重组腺病毒Ad-HBs,Ad-HBe,Ad- HBc,分别转染自脐带血体外诱导培养的DCs,观察腺病毒转染DCs效率和DCs中HBV抗原的表达；混合淋巴细胞反应(MLR)测定HBV抗原基因修饰DCs刺激同种异体T淋巴细胞增殖能力；乳酸脱氢酶释放法检测特异性CTL细胞对HepG_222．1．5靶细胞的杀伤能力．结果：腺病毒载体能够高效介导HBV三个抗原基因在DCs中表达,90%以上DCs表达示踪基因EGFP,且DCs细胞形态完整：感染后72 h HBsAg和HBeAg含量分别为0．919和0．328(吸光度A值)．MLR实验显示,HBV抗原基因修饰DCs仍然具有刺激同种异体T细胞的增殖能力,Ad-HBs转染DCs组、Ad-HBe转染DCs组、Ad-HBc转染DCs组和未转染DCs组之间刺激T细胞的增殖水平无明显差异(F=1．194,P=0．389)；在E：T比例为2：1,10：1和25：1时,Ad-HBs转染DC组、Ad-HBe转染DCs组和Ad-HBc转染DCs组对HepG_222．1．5细胞的杀伤率均明显高于未转染DCs组(P＜0．001)；以Ad- HBc转染DC组对HepG_222．1．5细胞杀伤率最高．结论：HBV抗原基因修饰DCs疫苗具有刺激同种异体T细胞增殖能力,同时能增强抗HBV特异性CTL反应的能力,可能发展为一种新型抗病毒疫苗．
文摘Hepatitis C virus(HCV) has a high propensity to establish chronic infections. Failure of HCV-infected individuals to activate effective antiviral immune responses is at least in part related to HCV-induced impairment of dendritic cells(DCs) that play a central role in activating T cell responses. Although the impact of HCV on DC phenotype and function is likely to be more prominent in the liver, major HCV-induced alterations are detectable in peripheral blood DCs(pb DCs) that represent the most accessible source of DCs. These alterations include numerical reduction, impaired production of inflammatory cytokines and increased production of immunosuppressive IL10. These changes in DCs are relevant to our understanding the immune mechanisms underlying the propensity of HCV to establish persistent infection. Importantly, the noninvasive accessibility of pb DCs renders the analysis of these cells a convenient procedure that can be serially repeated in patient follow-up. Accordingly, the study of pb DCs in HCV-infected patients during conventional treatment with pegylated interferon and ribavirin indicated that restoration of normal plasmacytoid DC count may represent an additional mechanism contributing to the efficacy of the dual therapy. It also identified the pre-treatment levels of plasmacytoid DCs and IL10 as putative predictors of response to therapy. Treatment of chronic HCV infection is changing, as new generation direct-acting antiviral agents will soon be available for use in interferon-free therapeutic strategies. The phenotypic and functional analysis of pb DCs in this novel therapeutic setting will provide a valuable tool for investigating mechanisms underlying treatment efficacy and for identifying predictors of treatment response.
基金Supported by National Natural Science Foundation of China,No.30871158 and No.81170604Outstanding Scientific Fund of Shengjing Hospital
文摘AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blot.Quantitative real-time polymerase chain reaction(PCR)was used to detect intestinal integrin-αm RNA expression.Intestinal CD83,CD86,CD74,CD3 and AKT were detected by immunohistochemistry,Western blot and PCR.Phosphorylated-AKT(p-AKT)was detected by immunohistochemistry and Western blot.RESULTS:In the FHF group[D-galactosamine(D-Galn)+lipopolysaccharide(LPS)group],the mice began to die after 6 h;conversely,in the D-Galn and LPS groups,the activity of mice was poor,but there were no deaths.Immunohistochemistry results showed that in FHF,the expression of CD11b/c(7988400±385941vs 1102400±132273,P<0.05),CD83(13875000±467493 vs 9257600±400364,P<0.05),CD86(7988400±385941 vs 1102400±13227,P<0.05)and CD74(11056000±431427 vs 4633400±267903,P<0.05)was significantly increased compared with the normal saline(NS)group.Compared with the NS group,the protein expression of CD11b/c(5.4817±0.77 vs 1.4073±0.37,P<0.05)and CD86(4.2673±0.69 vs 1.1379±0.42,P<0.05)was significantly increased.Itg-α(1.1224±0.3 vs 0.4907±0.19,P<0.05),CD83(3.6986±0.40 vs 1.0762±0.22,P<0.05)and CD86(1.5801±0.32 vs 0.8846±0.10,P<0.05)m RNA expression was increased significantly in the FHF group.At the protein level,expression of CD74in the FHF group(2.3513±0.52)was significantly increased compared with the NS group(1.1298±0.33),whereas in the LPS group(2.3891±0.47),the level of CD74 was the highest(P<0.05).At the gene level,the relative expression of CD74 m RNA in the FHF group(1.5383±0.26)was also significantly increased in comparison to the NS group(0.7648±0.22;P<0.05).CD3 expression was the highest in the FHF group(P<0.05).In the FHF,LPS and D-Galn groups,the expression of AKT at the protein and m RNA levels was elevated compared with the NS group,but there wasno statistical significance(P>0.05).The p-AKT protein expr
