慢性乙型病毒性肝炎患者外周血细胞CCR5及CD30的表达

目的为研究细胞趋化因子受体5(CCR5)及细胞分化抗原30(CD30 )作为1类辅助性T细胞/细胞毒性T细胞(Th1/Tc1)、2类辅助性T细胞/细胞毒性T细胞(Th2/Tc2)表面标志的可行性及辅助性T细胞(Th)/细胞毒性 T细胞亚群(Tc)在病毒性肝炎免疫发病机制中的作用.方法对20 例正常人群(NC)及20例慢性乙型病毒性肝炎(CHB)患者外周血单核细胞(PMBC)经植物血凝素(PHA)培养前后CCR5、CD30的表达水平进行了检测.结果①培养前,NC与CHB患者PMBC的CCR5及CD30的表达无明显的差别(P＞0.05),但两组CCR5的表达均高于CD30(P＜0.05) .②培养后,CHB患者CCR5高于NC(P＞ 0.05),而CD30则低于NC(P＜0.05).③培养后,NC的CCR5 表达较培养前明显的减少(P＜0.05),而NC及CHB患者的CD30的表达较培养前均明显的增加(P＜0.05);培养后,CHB患者CCR5的减少量低于NC,而 CD3 0的增加量少于NC(P＜0.05).④培养前CHB患者CCR5/CD30低于NC,而培养后高于 NC(P＜0.05),两组培养后CCR5/CD30均明显低于培养前(P＜ 0.05 ).结论①CCR5及CD30目前可作为Th及Tc细胞亚群相对特异性的细胞表面标志物;②CHB患者体内CCR5及CD30的异常表达或两者表达的失衡即Th/Tc细胞亚群的失衡与病毒性肝炎的发病机理可能存在一定的关系.     

IgA肾病患者的Th细胞亚群紊乱及其意义

目的探讨IgA肾病Th细胞亚群紊乱与发病机制、免疫紊乱程度、临床表现及病理类型之间的关系。方法以趋化因子CCR3和CCR5为辅助性T细胞Th2和Th1细胞特异性表面标志,用流式细胞术测定IgA肾病患者及健康对照组外周血CCR3 CD4 /CCR5 CD4 的T细胞亚群;并检验患者组的尿常规和肾功能。结果IgA肾病患者外周血CCR3 CD4 的T细胞亚群为(5.00±4.46)%,较对照组(1.56±0.73)%明显增多,CCR5 CD4 的T细胞亚群为(7.11±6.28)%,较对照组(13.85±3.58)%明显减少(P<0.01);免疫紊乱程度与临床表现及病理类型无相关性(P>0.05)。结论IgA肾病患者外周血Th2免疫应答增强、Th1免疫应答减弱,可能与该病的发病机制有关。     

复方虫草多糖脂质体口服液对慢性乙型肝炎细胞免疫的影响

目的:探讨复方虫草多糖脂质体口服液对慢性乙型肝炎细胞免疫的影响.方法:113例慢性乙型肝炎患者随机分为两组,治疗组57例,对照组56例,两组均给予门冬钾镁、肌苷、VitC等基础治疗,治疗组加用复方虫草多糖脂质体口服液,观察治疗前后外周血单个核细胞(PBMC)T细胞数量及各群比例关系及IL-2、IFN-γ含量.结果:治疗后治疗组PBMC CD4 T淋巴细胞数量及CD4 /CD8 比值较对照组明显增高(P＜0.01,P＜0.05),培养上清液IL-2、IFN-γ含量明显增高(P＜0.01,P＜0.05).结论:复方虫草多糖脂质体口服液能促进慢性乙肝患者PBMC产生CD4 T淋巴细胞、IL-2、IFN-γ,能明显增强慢性乙型肝炎患者的细胞免疫功能.     

外周血淋巴细胞计数在艾滋病患者HAART治疗中的变化及其对疗效的影响

目的 研究外周血淋巴细胞计数在艾滋病(AIDS)患者高效抗反转录病毒治疗(HAART)中的变化,分析其对于疗效的影响。方法 选取我院2018年6月至2019年9月收治的艾滋病患者120例作为观察组,给予HAART,选择同期100例门诊体检健康者作为对照组。检测并分析不同时段患者的CD4+T淋巴细胞和B淋巴细胞计数,分析治疗后两种计数的相关性,并利用受试者工作特征曲线(ROC)分析B淋巴细胞最佳治疗切入点。结果 (1)观察组治疗前后的CD4+T及B淋巴细胞计数均低于对照组(P 0.05),观察组患者治疗1、3、6及12个月后的CD4+T及B淋巴细胞计数均明显高于治疗前(P 0.05),且呈时间依赖性相关趋势;(2)与无效亚组比较,有效亚组治疗6个月、12个月的CD4+T淋巴细胞计数均明显升高(P 0.05),与无效亚组比较,有效亚组治疗前后的B淋巴细胞均明显升高(P 0.05),有效亚组及无效亚组的CD4+T淋巴细胞及B淋巴细胞计数均有高于治疗前(P 0.05),且呈时间依赖性相关趋势;(3)治疗12个月,观察组患者的ΔCD4+T淋巴细胞与ΔB淋巴细胞呈明显正相关(r=0.396,P 0.05);(4)ROC曲线显示,治疗前外周血B淋巴计数在101.28个/μl时,曲线下面积为0.853,治疗有效的敏感度为81.93%,特异度为83.47%。结论 HAART可明显提升艾滋病患者机体CD4+T淋巴细胞和B淋巴细胞计数,同时治疗前B淋巴细胞计数对艾滋病患者HAART治疗效果有明显影响。     

CD4+CD25+调节性T细胞在再生障碍性贫血中的表达及意义

目的:探讨再生障碍性贫血(再障)患者治疗前、后外周血T淋巴细胞亚群、CD4+CD25+T细胞的变化及其临床意义.方法:采用流式细胞术检测23例再障患者治疗前、后外周血T淋巴细胞亚群、CD4+CD25+T细胞的比例,并与健康对照组进行比较.结果:(1)再障患者治疗前CD4+,CD4+/CD8+,CD4+CD25+明显低于健康对照组(P＜0.01),CD8+细胞则明显升高(P＜0.05);重型再障组与慢性再障组比较CD4+CD25+降低,但差异不显著(P＞0.05),CD3+,CD4+,CD4+/CD8+,CD4+CD25+无统计学意义.(2)再障患者治疗后与治疗前相比,CD4+,CD4+/CD8+,CD4+CD25+明显升高(P＜0.01).(3)再障患者治疗有效者与无效者比较CD4+,CD4+/CDs+,CD4+CD25+明显升高(P＜0.01).结论:再障患者存在T淋巴细胞亚群的失调及CD4+CD25+T淋巴细胞的异常表达,且检测外周血T淋巴细胞亚群、CD4+CD25+T细胞有助于了解再障患者的免疫状态、疗效评价及预后判断.     

慢性乙型肝炎患者外周血CD4+CD25+high调节性T细胞对免疫状态的影响

目的 探讨CD4 CD25 high调节性T(Tr)细胞在慢性乙型肝炎(CHB)患者外周血的变化及其对免疫状态的影响.方法 用流式细胞仪技术分析比较42例CHB患者及20例正常健康人外周血中CD4 CD25 highTr细胞,同时检测CD4 T细胞和CD8 T细胞.结果 与健康对照组相比,CHB组CD4 CD25 highTr细胞百分率显著升高(P＜0.05),而CD4 T细胞显著下降(P＜0.05);CD8 T细胞虽有增加趋势,但无差异(P＞0.05).CHB组CD4 CD25 highTr细胞百分率与HBV DNA载量有一定的关系,但与血清ALT水平无相关(P＞0.05).结论 CHB患者外周血CD4 CD25 highTr细胞升高可能是导致机体免疫功能下降的原因之一.     

HIV进入宿主细胞的分子机制及相关药物研究进展

艾滋病(AIDS)是由人类免疫缺陷病毒(HIV)引起的恶性传染病，HIV进入宿主细胞环节涉及病毒表面糖蛋白(gp120和gp41)与细胞表面受体蛋白(CD4，CCR5和CXCR4等)的相互作用。现就HIV进入细胞的分子生物学机制以及目前针对病毒吸附与穿入设计的新型抗AIDS药物作一综述。     

健脾益气方对慢性乙型肝炎病毒携带者T细胞免疫功能的影响

目的 观察健脾益气方对慢性乙型肝炎病毒（HBV）携带者T细胞免疫功能的影响.方法 将60例HBV DNA阳性慢性HBV携带者（ASC）随机分成健脾益气方治疗组（A组）、常规治疗组（B组）各30例,用流式细胞仪检测外周血T淋巴细胞亚群水平,并与健康体检者（健康对照组,n=18）进行比较;ASC治疗24周后再次检测外周血T淋巴细胞亚群.结果 慢性乙型肝炎病毒携带者外周血T淋巴细胞CD4＋、CD8＋计数与对照组差异有统计学意义（P＜0.01）;健脾益气方治疗组治疗前后T淋巴细胞CD4＋、CD8＋计数及CD4＋/CD8＋比值差异有统计学意义（P＜0.05或P＜0.01）,与常规治疗组治疗24周后T淋巴细胞计数差异有统计学意义P＜0.05）.结论 慢性HBV携带者体内存在T淋巴细胞亚群失衡,健脾益气方能够改善慢性乙型肝炎病毒携带者的T细胞免疫功能.     

慢性乙型肝炎患者不同免疫状态下外周血T细胞表面程序性死亡受体1的表达

目的 检测慢性乙型肝炎(CHB)患者不同免疫状态下外周血CD4+、CD8+T细胞表面程序性死亡受体1(PD-1)表达,探讨其与血清HBV DNA载量、ALT之间的关系.方法 收集CHB患者免疫耐受期(A组,24例)、免疫活化期(B组,64例)、HBeAg阴性(C组,23例)和11例健康对照者(D组)外周血,采用流式细胞术检测CD4+、CD8+T细胞表面PD-1表达,以实时荧光定量PCR检测血清HBV DNA,同时检测ALT.结果 B、C组CD4+、CD8+细胞表面PD-1表达显著高于A、D组(P＜0.05).C组CD8+T细胞表面PD-1表达显著高于B组(P＜0.05).相关性分析发现,不同免疫状态下CHB患者CD4+、CD8+T细胞表面PD-1表达水平与HBV DNA载量及ALT均无相关性.结论 CHB患者外周血T细胞表面PD-1表达与机体免疫状态有关.肝脏炎症损伤是影响PD-1表达的重要因素,HBV DNA载量不是主要影响原因.     

不同CD4+T淋巴细胞基线值艾滋病HAART后免疫重建效果分析

目的 分析不同CD4+T淋巴细胞基线值的艾滋病HAART后免疫重建效果.方法 选取本院2004年1月至2015年12月收治的146例艾滋病患者予以回顾性分析,依据患者CD4+T淋巴细胞基线值的不同分为对照组(75例)与研究组(71例),观察并对比两组患者CD4+T淋巴细胞数的变化情况.结果 治疗后研究组3、6、12、24个月CD4+T淋巴细胞数值均比对照组高(P＜0.05);研究组3、6、12、24个月CD4+T淋巴细胞数值增幅幅度均比对照组大(P＜0.05).结论 CD4+T淋巴细胞计数高低将对免疫重建的速率产生直接直接影响,且后期相对较平缓,有利于促进患者免疫系统重建,进而提高其生存质量.     

Small molecule HIV entry inhibitors: Part I. Chemokine receptor antagonists: 2004 - 2010

INTRODUCTION: HIV/AIDS is one of the most devastating diseases in the world affecting > 40 million people worldwide. Morbidity and mortality from AIDS are significantly reduced due to the advent of highly active antiretroviral therapy (HAART). Long-term toxicity, emergence of drug resistant HIV strains and drug-drug interactions limit the effectiveness of HAART therapy. Chemokine receptor antagonists can provide drugs with lesser side effects and enhanced anti-HIV activity. Maraviroc, a chemokine co-receptor 5 (CCR5) antagonist from Pfizer, is already in clinical use. AREAS COVERED: This review covers patents and patent applications for small molecule CCR5 and CXC chemokine receptor 4 (CXCR4) antagonists published between 2004 and 2010 and related literature with a focus on recent developments based on lead generation and lead modification. The reader will gain information about the development of small molecule CCR5 and CXCR4 antagonists from the major pharmaceutical and biopharmaceutical companies. EXPERT OPINION: Several small lead molecules (CCR5 and CXCR4 antagonists) have been modified over this period for enhanced therapeutic activity and to obtain drug-like properties. CCR5 antagonists such as TBK-652 and TBK-220 from Tobira Therapeutics, and vicriviroc from Schering Plough showed a lot of promise in the developmental stage.     

Altered sensitivity of an R5X4 HIV-1 strain 89.6 to coreceptor inhibitors by a single amino acid substitution in the V3 region of gp120

The replication of several R5X4 strains is blocked by single CXCR4 inhibitors such as AMD3100 or T140 although the target cells express both CXCR4 and CCR5 in vitro. To identify which region(s) of the Env are involved in the increased sensitivity to CXCR4 inhibitors, we isolated a T140-escape mutant using R5X4 HIV-1 strain 89.6. An isolated mutant harbored a single amino acid substitution in the V3 region of the Env (arginine 308 to serine R308S). Luciferase-reporter HIV-1 pseudotyped with the mutant Env showed that the substitution conferred total resistance to CXCR4 antagonists but increased sensitivity to a CCR5 antagonist TAK-779 in the infection of the cells expressing both CCR5 and CXCR4. Analyses using the cells expressing a single coreceptor showed that the mutant Env predominantly and efficiently utilized CCR5 rather than CXCR4 while retaining R5X4 phenotype. These results indicated that the sensitivities of the R5X4 strain to coreceptor inhibitors were altered by a single amino acid substitution in the V3 region of gp120.     

HIV-1协同受体及其抑制剂研究进展

协同受体CCR5和CXCR4分别是嗜巨噬细胞性HIV-1和嗜T细胞性TIV-1侵入靶细胞的主要受体。CCR5抑制剂如TAK-779、SCH—C等，和CXCR4抑制剂如AMD3100、T22等，能分别与CCR5和CXCR4结合，从而阻断HIV-1侵入靶细胞。本文综述了HIV-1与CCR5和CXCR4的结合机制及其抑制剂的研究进展。     

输血感染HIV患者外周血T淋巴细胞亚群变化研究

目的研究输血感染的艾滋病患者和HIV携带者外周血T淋巴细胞亚群的表达。方法利用白细胞分类和流式细胞分析法对20例HIV携带者和20例AIDS患者外周血淋巴细胞中CD3＋,CD4＋,CD8＋的表达及绝对数进行检测。结果 AIDS患者组CD3＋淋巴细胞数较正常值显著下降（P〈0.05）;HIV感染组与艾滋病组的CD4＋T淋巴细胞数显著下降（P〈0.01）;CD8＋淋巴细胞数显著增高（P〈0.01）;CD4/CD8＋比值倒置。结论感染HIV后外周血CD4＋T淋巴细胞数显著下降,随病情进展下降更显著。     

Inhibition of CCR5-mediated infection by diverse R5 and R5X4 HIV and SIV isolates using novel small molecule inhibitors of CCR5: effects of viral diversity, target cell and receptor density

Highly active anti-retroviral therapy (HAART) has been very effective in reducing viral loads in human immunodeficiency virus (HIV)-1 patients. However, current therapies carry detrimental side effects, require complex drug regimes and are threatened by the emergence of drug-resistant variants. There is an urgent need for new anti-HIV drugs that target different stages of the replication cycle. Several synthetic small organic molecules that inhibit HIV infection by binding to the CCR5 coreceptor without causing cell activation have already been reported. Here, we have exploited a series of CCR5 antagonists to investigate their effects on diverse HIV and the simian counterpart (SIV) isolates for infection of a variety of cell types via different concentrations of cell surface CCR5. These inhibitors show no cross-reactivity against alternative HIV coreceptors including CCR3, CCR8, GPR1, APJ, CXCR4 and CXCR6. They are able to inhibit a diverse range of R5 and R5X4 HIV-1 isolates as well as HIV-2 and SIV strains. Inhibition was observed in cell lines as well as primary PBMCs and macrophages. The extent of inhibition was dependent on cell type and on cell surface CCR5 concentration. Our results underscore the potential of CCR5 inhibitors for clinical development.     

Targeting of Th1-associated chemokine receptors CXCR3 and CCR5 as therapeutic strategy for inflammatory diseases

CXCR3 and CCR5 are chemokine receptor that are predominantly expressed on the surface of Th1 polarized T cells. In a variety of human and experimental autoimmune diseases the enhanced expression of CXCR3 and CCR5 binding chemokine ligands is followed by the recruitment of CXCR3- and CCR5-positive T cells, indicating an important role for these chemokine receptors in T cell-mediated tissue damage. In this review, we summarize a number of in vivo studies available on the neutralization of CXCR3 and CCR5 in inflammatory disease, and specifically focus on the potential therapeutic effects of CXCR3 and CCR5 blockade in human autoimmune disease and organ transplantation.     

Rescue of HIV-1 long-time archived X4 strains to escape maraviroc

Entry of Human Immunodeficiency Virus type 1 (HIV-1) into target cells is mediated by the CD4 receptor and a coreceptor, CCR5 or CXCR4. Maraviroc interferes with HIV entry by binding the CCR5 coreceptor. Virological failure to maraviroc-containing regimens can occur through the emergence of resistance, or through tropism evolution and broadened coreceptor usage. In the latter case, the physiological relevance of minority strains is a major concern. Here we report a retrospective analysis of coreceptor-usage and evolution based on 454-ultra-deep-sequencing of plasma and Peripheral Blood Mononuclear Cell (PBMC)-derived envelope V3-loops, accounting for coreceptor usage, from a patient who failed a maraviroc-containing regimen through the emergence of X4 strains. The X4 maraviroc-escape variant resulted from recombination between a long time archived proviral sequence from 2003 (5'-portion, including the V3-loop) and the dominant R5 strains circulating in plasma at the time of maraviroc-treatment initiation (3'-portion). Phylogenetic analyses and BEAST modeling highlighted that an early diverse viral quasispecies underwent a severe bottleneck following reinitiation of HAART and repeated IL-2 cycles between 1999 and 2001, leading to the transient outgrowth and archiving of one highly homogeneous X4 population from plasma, and to the expansion in plasma of one PBMC-derived R5 strain. Under maraviroc selective pressure, the early, no longer detectable X4 strains archived in PBMC were partially rescued to provide X4-determinants to the main circulating strain.     

Rescue of HIV-1 long-time archived X4 strains to escape maraviroc

Entry of Human Immunodeficiency Virus type 1 (HIV-1) into target cells is mediated by the CD4 receptor and a coreceptor, CCR5 or CXCR4. Maraviroc interferes with HIV entry by binding the CCR5 coreceptor. Virological failure to maraviroc-containing regimens can occur through the emergence of resistance, or through tropism evolution and broadened coreceptor usage. In the latter case, the physiological relevance of minority strains is a major concern.Here we report a retrospective analysis of coreceptor-usage and evolution based on 454-ultra-deep-sequencing of plasma and Peripheral Blood Mononuclear Cell (PBMC)-derived envelope V3-loops, accounting for coreceptor usage, from a patient who failed a maraviroc-containing regimen through the emergence of X4 strains. The X4 maraviroc-escape variant resulted from recombination between a long time archived proviral sequence from 2003 (5′-portion, including the V3-loop) and the dominant R5 strains circulating in plasma at the time of maraviroc-treatment initiation (3′-portion). Phylogenetic analyses and BEAST modeling highlighted that an early diverse viral quasispecies underwent a severe bottleneck following reinitiation of HAART and repeated IL-2 cycles between 1999 and 2001, leading to the transient outgrowth and archiving of one highly homogeneous X4 population from plasma, and to the expansion in plasma of one PBMC-derived R5 strain. Under maraviroc selective pressure, the early, no longer detectable X4 strains archived in PBMC were partially rescued to provide X4-determinants to the main circulating strain.     

以趋化因子受体为靶点的抗HIV药物研究进展

趋化因子受体是HIV进入宿主细胞的共受体,特别是CCR5和CXCR4在HIV进入免疫细胞过程中起着重要作用。以趋化因子受体为靶点的新型抗艾滋病药物的设计与开发已成为抗艾滋病领域研究的一个热点课题。本文对近期抗HIV-1趋化因子受体(CCR5和CXCR4)拮抗剂的研究进展进行综述。