慢性乙型肝炎病毒感染者病毒YMDD的自然变异

目的 了解慢性HBV感染患者外周血HBV YMDD自然变异情况及其影响因素.方法 采用引物特异性实时荧光PCR法检测慢性HBV感染者外周血HBV YMDD变异情况,并对影响YMDD自然变异检出率的可能因素进行单因素及多因素分析.根据不同资料分别采用χ~2检验、Fisher's确切概率法、t检验、秩和检验及Logistic回归分析进行统计学处理. 结果在196例未经抗病毒治疗的慢性HBV感染者中,检出存在YMDD自然变异株感染者21例(10.70%),其中YVDD阳性20例,YIDD阳性例1变;变异毒株占总病毒株超过50%者1例,25%～500者5例,9%～25%者15例.B基因型HBV感染病例中YMDD变异株的检出率(20.00%,12/60)显著高于C基因型HBV感染病例(7.38%,9/122),χ~2=6.28,P<0.05.患者性别、年龄、HBeAg状态、HBVDNA载量、疾病状态、病毒感染时间对YMDD自然变异株的检出率无显著影响. 结论 在未经抗病毒治疗的慢性HBV感染者中存在HBV YMDD自然变异;YMDD自然变异的发生率与患者性别、年龄、HBeAg状态,HBV DNA载量、疾病状态、感染时间无显著相关性.B基因型较C基因型HBV更易出现YMDD自然变异.     

不同HBV感染转归人群外周血PD-1拷贝数分布研究*

目的 调查HBV感染不同转归人群外周血程序性死亡受体-1（PD-1）基因拷贝数（CN）分布频率的差异。方法 在300例HBV感染恢复者、437例慢性乙型肝炎（CHB）、249例肝硬化（LC）和123例肝细胞癌（HCC）患者,采用AccuCopy法检测外周血PD-1基因拷贝数。选择5个可能影响慢性HBV感染结局的指标,即HBeAg滴度、年龄、性别、HBV DNA和PD-1拷贝数,行Logistic回归分析,以发现影响感染结局的指标。结果 HBV感染恢复者、CHB、LC和HCC组外周血PD-1单倍体检出率分别为11.7%、9.6%、6.0%和10.6%,多倍体检出率分别为89.3%、90.4%,94%和89.4%,显示LC组多倍体检出率显著高于HBV感染恢复组或HCC组 （P<0.05）;以CHB组作为估计参数,经Logistic回归分析发现年龄和血清HBV DNA载量为影响疾病转归的独立危险因素。结论 本研究结果提示影响HBV感染转归的因素还是病毒本身和患者年龄,而外周血PD-1基因拷贝数变异检测的意义还有待于进一步研究。     

血清IL-17水平及其基因多态性与HBV感染临床转归之间的关系研究*

目的 探讨血清白介素（IL）-17水平和IL-17-197A/G基因单核苷酸多态性与乙型肝炎病毒（HBV）感染临床转归之间的关系。方法 2015年3月~2017年8月在我院就诊的乙型肝炎肝硬化40例,慢性乙型肝炎120例,无症状慢性HBV携带者60例和自限性HBV感染者80例。采用酶联免疫吸附试验检测血清IL-17水平,使用全自动生化仪检测血浆丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和总胆红素（TBIL）水平,采用多聚酶链反应结合荧光探针DNA扩增技术检测血清HBV DNA载量,使用单核苷酸检测试剂盒检测IL-17-197A/G基因单核苷酸多态性。结果 乙型肝炎肝硬化组血清IL-17水平显著高于慢性乙型肝炎组、无症状慢性HBV携带者组和自限性HBV感染组（F_3,296=102.8,P均<0.05）;慢性乙型肝炎组血清IL-17水平显著高于无症状慢性HBV携带者组和自限性HBV感染组（P均<0.05）;在乙型肝炎肝硬化组,血清高HBV DNA载量组（>1×10⁵ copies/mL）血清IL-7水平更高（t=5.1,P<0.05）,血清ALT>40 U/L组血清IL-7水平更高（t=10.7,P<0.05）;在慢性乙型肝炎组和乙型肝炎肝硬化组,血清AST>40 U/L组血清IL-7水平更高（t=24.5,P<0.05;t=22.4,P<0.05）,血清TBIL>20 μmol/L组血清IL-7水平更高（t=7.3,P<0.05;t=12.8,P<0.05）;肝硬化患者IL-17-197 AA、AG和GG基因型分别为75.0%、10.0%和15.0%,等位基因A分布频率为57.5%,G分布频率为42.5%,而慢性乙型肝炎患者则分别为25.8%、16.7%、57.5%和34.2%、65.8%,组间差异显著（P<0.05）,乙型肝炎肝硬化组以IL-17-197 AA基因型和A等位基因分布频率为主,慢性乙型肝炎组以GG基因型和G等位基因分布频率为主,HBV携带者组以AA基因型和A等位基因分布为主,自限性HBV感染者以GG基因型和G等位基因分布为主。结论 宿主免疫水平和遗传背景的相互作用决定了HBV感染的临床转归,IL-17在乙型肝炎病毒感染慢性化进程中发挥了重要作用。IL-17-197A/G位点的AA基因型和A等位基因可能是HBV易感的宿主基因,增加了HBV感染的不良转归。     

HIV/HCV合并感染者和HCV单纯感染者NS3/4A蛋白酶、 NS5A抑制剂的天然耐药变异分析

目的 通过对抗病毒治疗前HIV/HCV合并感染者和HCV单纯感染者HCV NS3/4A蛋白酶、NS5A抑制剂相关耐药位点进行检测,探讨上述患者天然耐药变异的差异。方法 收集2016年1月—2020年1月在广州医科大学附属市八医院住院或门诊就诊的246例HIV/HCV合并感染者和HCV单纯感染者的血清标本,使用Illumina二代测序平台进行测序,比较已在中国获批准的NS3/4A蛋白酶、NS5A抑制剂相关耐药变异在两组患者的差异,纳入分析的药物包括阿舒瑞韦/达拉他韦（ASV/DCV,基因1b型）,艾尔巴韦/格拉瑞韦（EBR/GZR,基因1b型）和格卡瑞韦/哌仑他韦（GLE/PIB,泛基因型）。符合正态分布的计量资料两组间比较采用t检验,不符合正态分布的计量资料两组间比较采用Mann-Whitney U检验。计数资料两组间比较采用χ²检验或Fisher精确检验。结果 本研究纳入的246例患者的血清样本中,239例（97.2%）成功进行PCR扩增并测序,包括102例HIV/HCV合并感染者和137例HCV单纯感染者。对ASV/DCV和EBR/GZR相关耐药变异分析结果提...     

ICAM-1基因多态性与HBV感染不同临床结局之间的关系

目的探讨细胞间黏附分子l（ICAM-1）基因多态性与HBV感染不同临床结局之间的相关性。方法应用病例．对照研究和聚合酶链反应-序列特异性引物法（PCR-SSP）检测118例慢性持续性HBV感染患者（包括无症状HBV携带者、慢性乙型肝炎、乙肝后肝硬化患者）和60例HBV急性自限性感染者的ICAM-1基因G241R（G／A）、K469E（A／G）两个位点的多态性，比较各组间基因型和等位基因频率，并对数据进行统计分析。结果①ICAM-l G241R（G／A）位点总GG基因型频率在HBV慢性持续性感染组高于急性自限性感染组，但差异无统计学意义（X^2=1．38，P〉0．05）。②ICAM-1 K469E（A／G）位点，进展性肝病组（慢性乙型肝炎和肝硬化）总KK基因型和总K等位基因的频率与无症状携带者组和自限性感染组相比显著增高（X^2=8．60，P〈0．05；X^2=5．07，P〈0、05），而在自限性感染和无症状携带者之间却无显著差异。结论携带ICAM-1 K469E KK基因型和K等位基因的患者容易进展成慢性乙型肝炎甚至肝硬化，可致慢性HBV感染患者病情进展。     

我国部分地区慢性HBV感染者HDV感染情况调查

目的 了解目前我国部分地区慢性HBV感染者丁型肝炎病毒(HDV)感染流行情况。方法 2021年3月—2022年6月从全国10个省市自治区收集3 131例慢性HBV感染者血清,用抗-HDV IgG酶联免疫试剂检测全部血清标本。对抗-HDV IgG阳性标本用巢式逆转录聚合酶链式反应(nRT-PCR)法检测HDV RNA。对HDV RNA阳性标本的nRT-PCR扩增产物测序后进行序列分析,确定HDV基因型。分析抗-HDV IgG阳性患者的临床特征。计量资料两组间比较采用Mann-Whitney U秩和检验。计数资料两组间比较采用χ²检验或Fisher精确检验。结果 3 131例慢性HBV感染者的抗-HDV IgG阳性率为0.70%(22/3 131),内蒙古自治区、新疆维吾尔自治区、北京市和湖南省慢性HBV感染者的抗-HDV IgG阳性率分别为1.81%(16/886)、0.88%(2/226)、0.28%(2/708)和1.00%(2/200),其中内蒙古自治区慢性HBV感染者抗-HDV IgG阳性率显著高于北京市(P=0.004),其余地区间比较差异均无统计学意义(...     

HBsAg水平对ALT正常的HBeAg阳性慢性HBV感染者肝脏炎症发生的预测价值

目的 探讨ALT正常且高病毒载量的HBeAg阳性慢性HBV感染者人群的肝炎发生情况及预测因素。方法 本研究临床注册号为NCT04032275。回顾性选取2008年10月—2015年5月ALT正常、高病毒载量、HBeAg阳性的慢性HBV感染者183例，根据其肝穿刺活检结果分为肝炎组和无肝炎组。计量资料两组间比较采用t检验或Mann-Whitney U检验。计数资料两组间比较采用χ²检验。使用单因素二元logistics回归分析预测因素，使用逐步后退法进行多因素二元logistics回归，使用受试者工作特征曲线（ROC）和约登指数分析截断值。结果 肝炎组37例（20.2%）,无肝炎组146例（79.8%）,肝炎组的男性比例低于无肝炎组（45.9%vs 68.5%,χ²=6.508,P=0.011）、AST高于无肝炎组[24（21.25～35.55） U/L vs 21.2（18.08～24.65） U/L,Z=-3.344,P=0.001]、HBsAg log值低于无肝炎组[4.4（4.28～4.49） vs 4.46（4.4～4.74）,Z=-...     

HBV相关慢加急性肝衰竭TBil水平的合理界值分析

目的 探讨HBV相关慢加急性肝衰竭（HBV-ACLF）患者的合理胆红素诊断阈值,以期更为精准地早期诊断。方法回顾性分析解放军总医院第五医学中心2008年9月—2018年9月收治的1232例HBV-ACLF患者,根据基线血清TBil水平分为A组（TBil<205.2μmol/L）和B组（TBil≥205.2μmol/L）,比较2组患者临床特征及28 d、90 d、1年及3年生存情况。计量资料两组间比较采用t检验或Mann-Whitney U秩和检验。计数资料组间比较采用χ²检验。使用Kaplan-Meier法分析2组患者的生存率,并应用log-rank检验进行比较。结果 2组间年龄（t=3.188,P=0.001）、男性（χ²=33.833,P<0.001）、肝衰竭分型（χ²=39.987,P<0.001）、WBC（Z=6.586,P<0.001）、HGB（Z=4.272,P<0.001）、PLT（Z=3.680,P<0.001）、Cr（Z=4.505,P<0.001）、TC（Z=...     

汉族人群乙型肝炎病毒感染者外周血IL-28B基因多态性研究*

目的 分析乙型肝炎病毒（HBV）感染者外周血IL-28B基因型和等位基因频率分布,并探讨其与疾病病程和进展的关系。方法 本研究纳入江苏籍汉族健康人群145例和453例HBV感染者,后者包括无症状HBV携带者（ASC）45例,慢性乙型肝炎患者（CHB）181例,肝硬化（LC）患者69例,肝细胞癌（HCC）患者79例,乙型肝炎肝衰竭（LF）患者79例,采用PCR法和直接测序法检测外周血IL-28B基因rs12979860和rs8099917多态性位点。采用Pearson x²检验对IL-28B rs8099917与rs12979860位点进行Hardy-Weinberg平衡检验,计量资料以（x±s）表示,计数资料采用例数表示。应用SPSS 17.0软件进行方差分析、x²检验和Binary Logistic回归分析。结果 IL-28B基因rs12979860位点有CC、CT和TT 3个基因型,LF患者CC型和C等位基因频率分别为96.2%和98.1%,显著高于健康人群的87.6%和93.1%（OR=0.257,95%CI=0.068~0.973,P=0.045;OR=0.255,95%CI=0.070~0.928,P=0.038）;IL-28B基因rs8099917位点有TT 、TG和GG 3个基因型,LC患者TT型和T等位基因频率分别为92.8%和96.4%,显著高于健康人群的86.2%和92.4%（OR=0.288,95%CI=0.087~0.948,P=0.041;OR=0.299,95%CI=0.096~0.926,P=0.036）。结论 江苏地区汉族人群IL-28B基因多态性与HBV感染后不同疾病表型相关,IL-28B基因rs12979860的C等位基因和IL-28B基因rs8099917的T等位基因可能是HBV感染后病情进展的影响因素。     

乙型肝炎病毒基因变异与免疫调节的关系

目的探讨乙型肝炎病毒（HBV）基本核心启动子（BCP）及前C基因变异与慢性乙型肝炎患者外周血CD4^＋CD25^＋调节性T细胞水平的关系。方法应用基因芯片技术检测HBV BCP及前C基因T1762、A1764、A1896位碱基变化;应用流式细胞仪检测外周血CD4^＋CD25^＋调节性T细胞表达水平。结果急性乙型肝炎、慢性活动性乙型肝炎、肝硬化、肝癌患者血清中HBV DNA T1762、A1764、A1896位碱基突变率分别为0、41.2%、69.4%、100%;急性乙型肝炎、慢性活动性乙型肝炎、肝硬化、肝癌患者外周血CD4^＋CD25^＋调节性T细胞水平高于正常对照;HBeAg阳性感染者高于HBeAg阴性感染者;变异株感染者低于非变异株感染者（P均〈0.05）。结论前C/C基因变异与HBV感染后肝病的慢性化及临床病情加重有关,与CD4^＋CD25^＋调节性T细胞的表达水平相关。     

Evidence for gene recombination in FCGR3 gene variants

Background and Objectives  The genes encoding the Fcγ receptors (FcγR) IIIa and IIIb ( FCGR3A and FCGR3B ) are clustered on chromosome 1 band q23–24 and exhibit allelic polymorphism. We investigated the molecular basis of additional new FCGR3 genomic variation.
Materials and Methods  A segment shared by FCGR3A and FCGR3B containing the polymorphic nucleotide positions 141, 147, 227, 266, and 277 in exon 3 was cloned and sequenced from genomic DNA of 30 donors and 3 bacterial artificial chromosome (BAC) clones. A mixture consisting of isolated FCGR3B * 2 - and FCGR3A - plasmids was cloned and sequenced as well. Additionally, nucleotide databases were screened for clones with variant FCGR3 sequences.
Results  A total of 12 FCGR3 variants defined by the polymorphic positions were detected in whole blood genomic DNA from 23 of 24 FCGR3B * 2 and/or FCGR3B * 3 positive donors, the DNA from two of three BAC clones and in the DNA mixture of isolated FCGR3B * 2 - and FCGR3A - plasmids.
Conclusion  Nucleotide exchanges of the variants were non-random and resulted from two alternative nucleotides present in one of the polymorphic position of the basic FCGR3 forms. Polymerase chain reaction (PCR) artefacts cannot be excluded as origin of new variants, but there is strong evidence that at least two variants are the result of a somatic recombination.     

FCGR3A and FCGR2A polymorphisms may not correlate with response to alemtuzumab in chronic lymphocytic leukemia

The in vivo mechanism of action of alemtuzumab (anti-CD52; Campath-1H) remains unclear. With rituximab, FCGR3A and FCGR2A high-affinity polymorphisms have been associated with clinical response in lymphoma but not in CLL, suggesting potential divergent mechanisms of action between these 2 diseases. Herein, we examined FCGR3A (V/V, n = 4; V/F, n = 10; F/F, n = 19) and FCGR2A (A/A, n = 5; H/A, n = 22; H/H, n = 6) polymorphisms in 36 patients with relapsed CLL who were treated with thrice-weekly alemtuzumab for 12 weeks to assess the potential influence these high-affinity FcgammaR receptor polymorphisms had on response to alemtuzumab. Response to alemtuzumab was similar regardless of FCGR3A polymorphism (V/V, 25%; V/F, 40%; F/F, 32%) or FCGR2A polymorphism (A/A, 40%; H/A, 32%; H/H, 33%). These findings indicate that FCGR3A and FCGR2A polymorphisms may not predict response to alemtuzumab in CLL. Future studies examining larger cohorts of alemtuzumab-treated patients with CLL will be required to definitively determine the predictive value of specific FCGR polymorphisms to treatment response.     

Fcγ receptor gene polymorphisms in Japanese patients with systemic lupus erythematosus: Contribution of FCGR2B to genetic susceptibility

Objective

Human low‐affinity Fcγ receptors (FcγR) constitute a clustered gene family located on chromosome 1q23, that consists of FcγRIIA, IIB, IIC, IIIA, and IIIB genes. FcγRIIB is unique in its ability to transmit inhibitory signals, and recent animal studies demonstrated a role for FcγRIIB deficiency in the development of autoimmunity. Genetic variants of FcγRIIA, IIIA, and IIIB and their association with systemic lupus erythematosus (SLE) have been extensively studied in various populations, but the results were inconsistent. To examine the possibility that another susceptibility gene of primary significance exists within the FcγR region, we screened for polymorphisms of the human FCGR2B gene, and examined whether these polymorphisms are associated with SLE.

Methods

Variation screening of FCGR2B was performed by direct sequencing and polymerase chain reaction (PCR)‐single‐strand conformation polymorphism methods using complementary DNA samples. Genotyping of the detected polymorphism was done using genomic DNA, with a specific genotyping system based on nested PCR and hybridization probing. Association with SLE was analyzed in 193 Japanese patients with SLE and 303 healthy individuals. In addition, the same groups of patients and controls were genotyped for the previously known polymorphisms of FCGR2A, FCGR3A, and FCGR3B.

Results

We detected a single‐nucleotide polymorphism in FCGR2B, (c.695T>C), coding for a nonsynonymous substitution, Ile232Thr (I232T), within the transmembrane domain. The frequency of the 232T/T genotype was significantly increased in SLE patients compared with healthy individuals. When the same patients and controls were also genotyped for FCGR2A‐131R/H, FCGR3A‐176V/F, and FCGR3B‐NA1/2 polymorphisms, FCGR3A‐176F/F showed significant association. Two‐locus analyses suggested that both FCGR2B and FCGR3A may contribute to SLE susceptibility, while the previously reported association of FCGR3B was considered to be secondary and derived from strong linkage disequilibrium with FCGR2B.

Conclusion

These results demonstrate the association of a new polymorphism of FCGR2B (I232T) with susceptibility to SLE in the Japanese.

     

Association of a transmembrane polymorphism of Fcγ receptor IIb (FCGR2B) with systemic lupus erythematosus in Taiwanese patients

Objective

To investigate the possible association of the Fcγ receptor IIb (FcγRIIb) Ile/Thr187 transmembrane domain polymorphism, which significantly affects receptor signaling, with susceptibility to systemic lupus erythematosus (SLE) in Taiwanese patients.

Methods

We used matrix‐assisted laser desorption ionization−time‐of‐flight mass spectrometry to genotype 351 Taiwanese SLE patients and 372 age‐ and sex‐matched healthy individuals from the same geographic area. Allele frequencies and genotype distributions were compared between the patients and controls, both as an aggregate and as stratified by sex, autoantibody profile, and clinical parameters. A combined analysis was conducted to assess the FCGR2B Thr187 allele as a common risk factor in different ethnic populations.

Results

The minor Thr187 allele was significantly associated with SLE in Taiwanese subjects (P = 0.017, odds ratio [OR] 1.989 [95% confidence interval (95% CI) 1.119–3.553]). Interestingly, male SLE patients showed enrichment of the Thr/Thr187 genotype (24%; 7 of 29) as compared with female SLE patients (10%; 32 of 322) (P = 0.043, OR 2.884 [95% CI 1.028–7.839]). Additionally, SLE patients with Thr/Thr187 and Ile/Thr187 genotypes were more likely to have pleural effusions (P = 0.038, OR 1.874 [95% CI 1.033–3.411]) and anti‐SSA/Ro antibody production (P = 0.046, OR 2.221 [95% CI 1.013–4.897]). Combined analysis of 4 groups of Asian patients strongly supported the association of the FCGR2B Thr187 allele with the lupus phenotype (P = 0.000159).

Conclusion

The FcγRIIb transmembrane polymorphism is a strong disease susceptibility candidate in epistasis with other genetic effects in Taiwanese and other Asian populations. It may also play a more prominent role in male patients with SLE.