程序性细胞凋亡基因单核苷酸多态性与慢性乙型肝炎患者干扰素早期病毒学应答的相关性

Objective To investigate the relationship between the single nucleotide polymorphisms (SNP) of programmed cell death-1 (PD-1) gene and early virologic response of interferon-α (IFN-α) in patients with chronic hepatitis B (CHB). Methods A total of 135 CHB patients were prospectively enrolled in this study. SNP of PD-1.1 and PD-1.2 genes were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in these patients.Then its relationship with early virologic response to IFN-α treatment was analyzed. The data were analyzed by x2 test. Results Among the 135 patients, 33 (24.4%) achieved early virologic response to IFN-α. There were 35, 77, and 23 patients with AA, AG, GG genotgpe of PD-1.1. The early virologic response was achived in 5(14.3%), 25(32.5%) and 3(13.0%) among patients with AA,AG, GG genotypes of PD-1.1, respectively. There were statistically different (x2 = 6. 258, P =0. 044). The subjects with AG genotype showed higher response rate than those with AA or GG genotypes (x2 = 6. 246, P= 0. 012). However, the early virologic response rates were not significant different among subjects with AA, AG or GG genotype of PD-1. 2 ( x2= 3.957, P= 0. 138).Conclusion SNP of PD-1.1 gene may be used as a marker to predict the early virologic response to IFN-α treatment in Chinese CHB patients.     

程序性细胞凋亡基因单核苷酸多态性与慢性乙型肝炎患者干扰素早期病毒学应答的相关性

目的 探讨程序性细胞凋亡基因-1(PD-1)单核苷酸多态性(SNP)与IFN-α治疗慢性乙型肝炎(CHB)患者发生早期病毒学应答的关系.方法 采用前瞻性队列研究方法,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术,检测135例中国汉族抗病毒初治CHB患者的PD-1.1和PD-1.2的SNP,并分析其与IFN-α早期病毒学应答的关系.分类变量资料采用x2检验.结果 135例CHB患者IFN-α治疗获得早期病毒学应答有33例,占24.4%.PD-1.1的AA、AG、GG基因型各占35、77和23例,IFN-α的早期病毒学应答各有5、25和3例,分别占14.3%、32.5%和13.0%(x2＝6.258,P＝0.044),AG基因型相对AA、GG基因型有较高的病毒学应答率(x2＝6.246,P＝0.012).在PD-1.2的AA、AG、GG基因型之间比较IFN-α的早期病毒学应答,差异无统计学意义(x2＝3.957,P=0.138).结论 PD-1.1的SNP与中国汉族CHB患者IFN-α治疗早期病毒学应答有关.

Abstract：

Objective To investigate the relationship between the single nucleotide polymorphisms (SNP) of programmed cell death-1 (PD-1) gene and early virologic response of interferon-α (IFN-α) in patients with chronic hepatitis B (CHB). Methods A total of 135 CHB patients were prospectively enrolled in this study. SNP of PD-1.1 and PD-1.2 genes were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in these patients.Then its relationship with early virologic response to IFN-α treatment was analyzed. The data were analyzed by x2 test. Results Among the 135 patients, 33 (24.4%) achieved early virologic response to IFN-α. There were 35, 77, and 23 patients with AA, AG, GG genotgpe of PD-1.1. The early virologic response was achived in 5(14.3%), 25(32.5%) and 3(13.0%) among patients with AA,AG, GG genotypes of PD-1.1, respectively. There were statistically different (x2 = 6. 258, P =0. 044). The subjects with AG genotype showed higher response rate than those with AA or GG genotypes (x2 = 6. 246, P= 0. 012). However, the early virologic response rates were not significant different among subjects with AA, AG or GG genotype of PD-1. 2 ( x2= 3.957, P= 0. 138).Conclusion SNP of PD-1.1 gene may be used as a marker to predict the early virologic response to IFN-α treatment in Chinese CHB patients.     

慢性乙型肝炎患者干扰素-α治疗应答与干扰素-α/β受体启动子-408基因多态性的相关性

目的:探讨干扰素(IFN)-α/β受体启动子-408位点的单核苷酸多态性(SNP)与IFN疗效的关系。方法:应用聚合酶链反应(PCR)并基因测序法检测210例慢性乙型肝炎(CHB)患者(轻度38例、中度102例、重度70例)的IFN-α/β受体基因启动子-408位点的SNP。对210例中既往未用过IFN治疗的80例CHB患者给予聚乙二醇化干扰素α-2a or 2b(Peg-IFN-α-2a/2b)治疗48周。比较SNP与Peg-IFN疗效的关系。结果:80例患者中,IFN治疗持久应答(SR)33例(41.3%),非持久应答(NSR)47例(58.7%)。IFN-α/β受体启动子-408位点CT基因型患者干扰素治疗SR为65%(13/20),显著高于CC基因型患者干扰素治疗SR 29.5%(13/44),两者比较差异有显著性意义(χ2=7.166,P=0.007)。干扰素治疗SR组IFN-α/β-408位点等位基因C、T的频率与NSR组的频率比较差异无显著性意义(P>0.05)。结论:IFN-α/β受体启动子-408为CT杂合基因型的CHB患者可能对Pec-IFN治疗效果较好。     

干扰素-γ基因多态性与干扰素治疗慢性乙肝患者持久应答的关系

目的探讨干扰素-γ(IFN-γ)启动子基因874位点的单核苷酸多态性(SNP)与IFNα-2b治疗慢性乙肝(CHB)持久应答的关系。方法选择CHB患者106例,应用PCR-SSP技术分析宿主的IFN-γ启动子基因874位点的SNP,患者给予IFNα-2b治疗1年,随访1年,比较SNP与IFNα-2b疗程结束时完全应答、停药后随访1年完全应答(持久应答)的关系。结果在标准疗程结束时,完全应答的患者中三种基因型的分布无统计学差异(χ2=3.594 9,P=0.165 7)。而在持久应答患者中三种基因型的分布有统计学差异,TT基因型患者的持久应答率高于其他两种基因型患者(χ2=6.639 8,P=0.036 1)。结论 CHB患者对IFNα-2b治疗的持久应答与IFN-γ基因型有一定关联性,尤其与TT基因型关联更大。     

干扰素α抗病毒治疗效果与人白细胞抗原-DRB1*11基因单核苷酸多态性的相关性

目的 探讨人白细胞抗原-DRB1*11基因片段内5个干扰素α调节区间单核苷酸多态性的分布与慢性乙型肝炎患者对干扰素α治疗效果的关系. 方法 采用回顾性分析研究方法 ,随机抽取107例慢性乙型肝炎患者在我院经过干扰素α治疗12个月,已停药6个月的患者,分持续应答组(A组)和非持续应答组(B组).在美国国立生物技术信息中心上查询人乙型肝炎病毒X蛋白结合蛋白基因位点,使用DNA池技术测序作单核苷酸多态性位点的验证,针对各个单核苷酸位点分别设计引物和TaqMan-MGB探针,检测5个干扰素α调节区的单核苷酸多态性,分析两组间的差异.结果 107例慢性乙型肝炎患者对干扰素a持续应答的30例,非持续应答的77例,持续应答组5个干扰素α调节区中基因型CT的占有率为18.0%,AG为10.8%,非持续应答组基因型CT的占有率为23.8%,AG为15.8%,持续应答组CT、AG这两对基因型的占有率比非持续应答组低,两组比较,x2值分别为7.728和7.956,P＜0.05,差异有统计学意义,其余基因型之间比较,差异无统计学意义.结论 检测干扰素调节基因,可粗略评估患者对干扰素应答的状况,有助于临床医师预测患者对干扰素α的疗效.     

慢性乙型肝炎患者抗前S1蛋白C末端IgG与干扰素早期应答的相关性

目的 检测B基因型HBV感染慢性乙型肝炎患者血清中抗前S1蛋白C末端(94～117 aa)IgG抗体(简称抗S1),探讨其与干扰素治疗早期应答的关系.方法 采用型特异性引物PCR方法进行基因型测定.对69例使用干扰素治疗的B基因型慢性乙型肝炎患者进行前瞻性研究,其中使用普通干扰素(α-1b、α-2b)42例,聚乙二醇干扰素α-2a 27例,按照HBV B基因型序列合成前S1蛋白C末端(94～117 aa)多肽,用酶联免疫吸附法检测其相应抗体的存在情况.干扰素早期应答以随访12周时的病毒载量下降幅度、转氨酶和HBV血清学标志物的变化为指标进行评价.根据抗S1的反应性将69例B基因型HBV感染的慢性乙型肝炎患者分为抗S1阳性组和抗S1阴性组,评判两组干扰素疗效的差别.数据分析采用SPSS13.0统计软件,两组间计量数据应用t检验或Mann-Whitney U检验,计数数据采用χ~2检验、Fisher's确切概率法或Mann-Whitney U检验,P<0.05为差异有统计学意义.结果 抗S1阳性组21例,阴性组48例,抗S1阳性和阴性组间干扰素种类差异无统计学意义.治疗12周时,抗S1阳性组和阴性组HBV DNA下降水平平均分别为3.37log_(10)拷贝/ml和0.33log_(10)拷贝/ml,Z=-3.658,P=0.000;ALT下降水平平均分别为92 U/L和30.5U/L,Z=-2.132,P=0.033,差异均有统计学意义.治疗12周时,抗Sl阳性组和阴性组HBeAg阴转率分别为41.2%(7/17)和5.9%(1/17);血清学转换率分别为12.8%(5/39)和0(0/39),Z=-5.11,P=0.000;病毒学应答率分别为71.4%(15/21)和16.7%(8/48),χ~2=19.710,P=0.000;完全应答率分别为23.8%(5/21)和6.2%(3/48),部分应答率分别为52.4%(11/21)和18.8%(9/48),Z=-4.840,P=0.000,差异均有统计学意义.抗S1在预测干扰素的早期应答中阳性预测值为71.4%,阴性预测值则为83.3%.结论 通过对治疗前血清中抗S1的检测,可以筛选出合适的慢性乙型肝炎患者,进行个体化治疗,提高干扰素治疗的应答率.     

重组α干扰素治疗慢性乙型肝炎病毒学反弹与干扰素中和抗体的相关性研究

目的 观察和分析重组α干扰素（rIFNα）治疗CHB过程中出现病毒学反弹与干扰素中和抗体（NAb）产生的关系。方法 485例经肝穿刺活组织检查证实的CHB患者,给予rIFNα1b治疗,每次5×106U,每周3次,疗程个体化（6-37个月,中位数10个月）,疗程中1-3个月检测1次肝功能,并以PCR荧光定量测定HBV DNA、Abbott EIA试剂检测HBeAg、抗病毒生物中和法检测NAb。结果 485例患者中,66例（13．6％）于治疗过程中出现病毒学反弹,NAb阳性98例（20．2％）,病毒学反弹组NAb阳性率68．2％（45／66）,显著高于未发生病毒学反弹组的12．6％（53／415）,差异有统计学意义（x^2=109．06,P〈0．01）。NAb阳性组病毒学反弹的发生率为45．9％（45／98）,显著高于NAb阴性组的5．4％（21／387）,差异有统计学意义（x^2=109．06, P〈0．01）。病毒学反弹的时间与NAb检出的时间存在相关性及一致性（CR=98．3％,P〈0．01）。多因素分析表明,NAb的检出是病毒学反弹发生的相关因素之一（P〈0．01）,患者的年龄和治疗前HBeAg状态亦可影响病毒学反弹的发生。结论 rIFNα治疗CHB过程中病毒学反弹的发生率约13．6％,病毒学反弹的发生与NAb的产生密切相关,患者的年龄和治疗前HBeAg水平亦可影响病毒学反弹的发生。     

慢性乙型肝炎患者干扰素α受体1基因启动子多态性和干扰素应答的关系

目的 探讨慢性乙型肝炎患者的Ⅰ型干扰素受体1(IFNAR1)基因启动子多态性和IFN-α治疗应答之间的关系.方法 选择接受IFN-α治疗的慢性乙型肝炎患者61例,采用重组IFN-α2b 500万U,隔天肌内注射,疗程48周,观察应答情况,对入选患者的IFNAR1基因启动子区进行测序,计量资料采用t检验,计数资料采用卡方检验.结果 治疗的慢性乙型肝炎患者中,完全应答22例,部分应答8例,无应答31例.IFNAR1基因启动子区一408C/T、-3C/T、-77GT双核苷酸重复序列[-77(GT),]存在基因多态性,这三个位点基因多态性存在连锁,形成-408C/-77(GT)5/-3C等基因单体型.IFNAR1启动子区基因型为-408C/-77(GT)5/-3C及-408C/-77(GT)5/-3C的,基因型为-408C/-77(GT)5/-3C和非-408C/-77(GT)5/-3C的慢性乙型肝炎患者对IFN-α的应答率为61.0%,高于基因型为非-408C/-77(GT)5/-3C,非-408C/-77(GT)5/-3C患者的25.0%(X2=6.961,P=0.008).结论 IFNAR1启动子基因型为-408C/-77(GT)5/-3C及-408C/-77(GT)5/-3C的,-408C/-77(GT)5/-3C和非-408C/-77(GT)5/-3C的慢性乙型肝炎患者对IFN-α治疗应答较好,IFNAR1基因启动子多态性与慢性乙型肝炎患者的干扰素应答有关.     

慢性丙型肝炎患者Ⅰ型干扰素受体启动子基因多态性与抗病毒疗效的关系

目的研究慢性丙型肝炎(CHC)患者Ⅰ型干扰素受体之一IFNAR1启动子基因的单核苷酸多态性与干扰素(IFN)α抗HCV疗效的关系。方法应用聚乙二醇干扰素α联合利巴韦林抗HCV治疗,根据抗病毒疗效将73例患者分为应答组与无应答组,并对其IFNAR1启动子基因进行扩增及测序。结果 73例患者中应答组47例,无应答组26例,在-568、-408、-77、-3位点发现基因多态性,共10种基因单体型,其中单体型-568C/-408C/-77(GT)5/-3C和-568C/-408T/-77(GT)5/-3T出现频率最高。非-568C/-408C/-77(GT)5/-3C者的抗病毒应答率77.1%高于-568C/-408C/-77(GT)5/-3C者52.6%,(χ2=4.773,P=0.029),-568C/-408T/-77(GT)5/-3T者的抗病毒应答率90.9%高于非-568C/-408T/-77(GT)5/-3T者59.7%,(χ2=3.92,P=0.048)。结论 CHC患者IFNAR1启动子基因多态性可能影响IFNα抗HCV治疗应答。     

慢性乙型肝炎病毒感染者转化生长因子-β1基因单核苷酸多态性分析

目的 探讨转化生长因子(TGF)-β1单核苷酸多态性(SNP)与慢性HBV感染的关系.方法 采用PCR方法,对74例慢性乙型肝炎患者、41例乙型肝炎肝硬化患者和41例健康人外周血细胞基因组DNA的TGF-β1基因中三个SNP位点(rs2241715、rs2241716、rs4803455)进行基因分型.数据分析采用χ2或Fisher精确概率法检验.结果 健康人的 rs2241715基因型及等位基因频率与慢性乙型肝炎和肝硬化患者相比,差异有统计学意义(χ2=11.419,P＜0.01或χ2=6.218χ2=5.961;P＜0.05),rs2241715 T/T基因型的个体患慢性乙型肝炎和乙型肝炎肝硬化的相对危险度分别为其他型的2.974倍(95%CI=1.209～7.314,P=0.018)和3.228倍(95%CI=1.201～8.675,P=0.020),rs2241716和rs4803455位点则无上述相关性.结论 TGF-131基因的rs2241715 SNP位点T/T基因型可能与慢性HBV感染密切相关.     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎病毒感染者程序性死亡因子1基因拷贝数和mRNA的表达水平

目的 研究慢性HBV感染者程序性死亡因子1(PD-1)基因的拷贝数和mRNA表达水平,以及在不同临床表现患者间的差异。 方法 实时荧光定量PCR法检测健康对照者27例（对照组）、慢性HBV携带者31例（携带组）、慢性重型肝炎患者19例（重肝组）、乙型肝炎相关性原发性肝癌患者29例（肝癌组）的外周血单个核细胞PD-1基因...     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

目的 研究程序性死亡分子-1(PD-1)及其配体(PD-L1)表达水平与慢性HBV感染者HBV DNA水平的相关性及抗病毒治疗对其表达的影响.方法 检测137例慢性HBV感染者的外周血单个核细胞(PBMC)表面PD-1和PD-L1,并检测其中64例人类白细胞抗原(HLA)-A2阳性者HBV特异性CTL数量.ELlSA法检测PBMC体外培养上清液中IFN-γ浓度.比较10例HBeAg阳性慢性乙型肝炎(CHB)患者予替比夫定抗病毒治疗24周前后上述指标的变化.两组间均数比较采用两独立样本的t检验,多组间的差异采用单因素方差分析,相关分析采用Pearson相关分析.结果 HBV DNA＜3 lg、3～6 lg和＞6 lg拷贝/mL组问PBMC表面PD-1和PD-L1表达均明显高于健康对照组,但差异无统计学意义;3组HBV特异性CTL表面PD-1表达分别为(69.3±11.2)%、(76.5±9.1)%和(78.0±11.7)%,HBV DNA＞6 lg拷贝/mL 组PD-1表达明显高于＜3 lg拷贝/mL组,而HBV特异性CTL数量明显低于＜3 lg拷贝/mL组;3组PBMC体外培养上清液中IFN-γ水平差异无统计学意义.HBeAg阳性组和阴性组间上述指标差异无统计学意义.替比夫定抗病毒治疗12周和24周时,PD-1、PD-L1表达较治疗前明显下降,伴有HBV特异性CTL数量逐渐增加和IFN-γ水平升高.结论 慢性HBV感染者PBMC表面PD-1的表达较健康者明显上调,且HBV特异性CTL表面表达PD-1水平与血清HBV DNA水平相关,但与HBeAg状态无关.抑制HBV复制能降低PD-1、PD-L1表达,并增加HBV特异性CTL的数量和功能.

Abstract：

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

主要组织相容性复合体Ⅱ类反式激活因子基因编码区单核苷酸多态性与慢性乙型肝炎病毒感染的转归

目的 探讨主要组织相容性复合体(MHC)Ⅱ类反式激活因子(CⅡTA)编码区两个非同义单个核苷酸多态性(SNP)位点C19170G、C30799G的多态性与慢性HBV感染不同临床表型的关系.方法 在627例不同临床表型的慢性HBV感染人群与101名健康献血员中,用序列特异性引物-聚合酶链反应(PCR-SSP)的方法对CⅡTA编码区两个非同义SNP位点C19170G、C30799G进行基因分型;X2检验判断上述位点的基因型分布是否符合Hardy-Weinberg平衡;列联表X2检验检测两组间的差异;非条件Logistic回归进行分析.结果 CⅡTA基因编码区19170位点G等位基因和GG+GC基因型在肝硬化人群中的频率显著高于其在非进展性肝病人群中的频率(X27.128,P=0.008;X26.404,P=0.011),且各基因型频率在肝硬化与非进展性肝病两组人群间存在显著差异(OR:0.742,95%CI:0.552～0.998,P=0.048),其两者间的差异主要存在于G基因显性模式下(OR:0.581,95%CI:0.353～0.954,P=0.032);30799位点C等位基因和CC基因型在肝细胞癌人群中的频率显著高于其在肝硬化人群中的频率(X24.861,P=0.027;X24.993,P=0.025).且各基因型频率在肝硬化与肝细胞癌两组人群间存在显著差异(OR:0.557,95%CI:0.334～0.930,P=0.025),其两者间的差异主要存在于C基因隐性模式下(OR:0.491,95%CI:0.269～0.898,P=0.021).结论 在慢性HBV感染者中,C19170G位点多态性与肝硬化的形成密切相关,携带G等位基因者易发展为肝硬化;C30799G位点多态性与肝细胞癌的发生密切相关,携带CC基因型者易发展为肝细胞癌.     

B7-H1及其受体PD-1在慢性乙型肝炎患者淋巴细胞上的表达及意义

目的研究B7-H1及其受体PD-1在慢性乙型肝炎患者T淋巴细胞及髓样树突细胞（mDCs）上的表达及它们的表达水平与患者疾病状态的关系。方法流式细胞技术检测正常人和慢性乙型肝炎患者CD4^＋、CD8^＋T淋巴细胞及mDCs上B7-H1和PD-1的表达水平。实时定量PCR检测患者的HBV DNA。结果慢性乙型肝炎患者B7-H1及其受体PD-1的表达水平明显升高,健康对照mDCs、CD4^＋及CD8^＋T淋巴细胞B7-H1的阳性表达率分别为0.35%±0.10%、3.63%±0.70%和1.20%±0.19%,慢性乙型肝炎患者分别为7.88%±1.40%、24.28%±2.86%和10.78%±1.62%,慢性乙型肝炎患者B7-H1在mDCs和T淋巴细胞上的表达水平明显高于健康对照（P值均〈0.05）健康对照CD4^＋及CD8^＋T淋巴细胞PD-1的阳性表达率分别为5.92%±1.75%和5.98%±0.88%,慢性乙型肝炎患者分别为17.76%±2.47%和11.92%±2.21%,慢性乙型肝炎患者PD-1在T淋巴细胞上的表达水平也明显高于健康对照（P值均〈0.05）。且它们的表达与患者的ALT水平及HBV DNA载量呈明显的正相关（P值均〈0.05）。结论慢性乙型肝炎患者淋巴细胞上B7-H1和PD-1的表达水平与患者疾病状态密切相关。