Development of immunotherapeutic strategies for HIV-1

In the majority of untreated patients, HIV-1 infection presents as a progressive disease of the immune system. Recent studies indicate that immune responses can be induced in HIV-1 infected individuals, leading to some immune control of virus replication. Such immune responses are also observed in small numbers of untreated HIV-1 infected long-term non-progressor (LTNP) patients, as well as in other viral infections (including those with human herpesviruses). Emerging novel technologies, animal studies and detailed immunological studies have proven invaluable in defining the immune responses that are associated with a favourable clinical outcome. Central effector and regulatory cells are HIV-1-specific CD8+ cytotoxic T-lymphocytes (CTL) and CD4+ helper T-lymphocytes respectively. Fully functional antigen-presenting cells (APC) are also essential in all stages of HIV-1 infection and possibly some (but not all) antibody responses contribute to beneficial immunity. The availability of combination anti-retroviral drug therapy, which successfully controls viraemia, has enabled a beneficial outcome in many HIV-1 infected individuals. Since no chronically HIV-1 infected patient has been shown to eradicate virus, novel approaches utilising therapeutic immunisation and various cytokines to manipulate immune responses and to induce and steer immunity towards a desired phenotype are required. There is a clear rationale for immunotherapeutic intervention in chronic progressive HIV-1 infection, which forms the foundation for novel approaches aimed at inducing and maintaining immune control. Here we review the immunopathogenesis of HIV-1 infection and discuss the promises of therapeutic immunisation and immunotherapy in general and their potential in the treatment of chronic HIV-1 disease.     

HIV/AIDS患者152例CD4+细胞计数与机会性感染相关分析

目的:分析HIV/AIDS患者CD4+细胞计数与机会性感染的关系,为HIV/AIDS患者机会性感染的治疗及一、二级预防提供参考依据.方法:对2009年6月至2011年2月在我院住院的152例HIV/AIDS患者的CD4+细胞及出现的机会性感染进行分析.按患者CD4+细胞计数分为CD4+＞200个/μuL与CD4+≤200个/μL两组,比较两组患者机会性感染发生率的差异.结果:152例HIV/AIDS患者机会性感染的总感染率为93.4%,主要的机会性感染是口腔念珠菌感染(57.2%)、肺结核(50.7%)、细菌性肺炎(32.9%)、败血症(22.4%)、感染性腹泻(21.1%).CD4+细胞计数≤200个/trL组患者机会性感染的发生率为96.0%,高于CD4+细胞计数＞200个/μL组患者(80.8%),差异有统计学意义(P＜0.05);随着CD4+细胞计数的下降,患者发生机会性感染的几率增高.结论:HIV/AIDS患者机会性感染的发生率高,CD4+细胞计数是HIV/AIDS患者发生机会性感染的独立危险因素.因此,应定期监测HIV/AIDS患者CD4+细胞计数,加强患者机会性感染的一级和二级预防.     

大理市HIV感染者中HCV感染状况的研究

目的探讨人类免疫缺陷病毒(HIV)-1感染者中丙型肝炎病毒(HCV)的混合感染率,并了解HCV对HIV-1感染者CD4+T细胞计数的影响。方法采用横断面研究方法,在云南大理市招募HIV-1感染者,分别采用血清学和核酸方法检测HCV混合感染。结果在526例HIV-1感染者中,静脉吸毒者占94.3%,其余为性途径感染。86.9%(457/526)为HCV血清学检测抗体阳性,其中HCV核酸阳性者占78.3%。在HCV血清学阴性的69例中,24例HCV RNA>1 000 IU/mL。由于引入HCV核酸检测方法,现场HCV混合感染的发现率增加了4.6%(24/526),所调查的HIV-1感染者中HCV混合感染率为91.4%。HCV混合感染者的CD4+T细胞计数明显低于HIV-1单纯感染者。结论在HCV感染的高流行区或髙危人群中,HCV与HIV-1共感染率较高。筛查HIV-1感染时应加强对HCV的检测,有助于对HCV感染进行早期诊断并开展HCV的早期治疗,减少HCV对HIV-1感染者的不利影响。     

Breaking the asymptomatic phase of HIV-1 infection

AIDS typically consists of three phases: (1) an acute, infectious mononucleosis-like syndrome followed by (2) a prolonged asymptomatic stage ending in (3) the appearance of frank AIDS. The asymptomatic phase may last for years and its presence suggests a persistent conflagration between the virus and the host's immune response. There is considerable evidence that an immune response develops but the response is ultimately inadequate. From the work of others as well as our own, we have constructed a hypothesis which attempts to explain some aspects of the immune response. We propose that HIV-1 preferentially infects a subset of CD4⁺ lymphocytes which are then either destroyed or altered in their biological functions. Further, we suggest that this subset represents the CD4⁺ TH1 lymphocyte population. By decreasing the quantity of IL-2 and interferon-gamma produced by TH1 lymphocytes, the production of cytokines by TH2 cells is increased. One of the cytokines produced by TH2 lymphocytes is IL-10, a polypeptide with significant inhibitory properties towards lymphocytes. Sera from patients with frank AIDS have significant lymphocyte inhibitory activities some of which operate through IL-10. Thus, a gradual shift to a TH2-type response and release of increasing amounts of inhibitors eventually prevents the host from replacing destroyed cells or mounting new and appropriate immune responses. © 1994 Wiley-Liss, Inc.     

2011年新报告的艾滋病病毒感染者接受抗病毒治疗前CD4+ T淋巴细胞计数自然变化情况分析

目的 探讨2011年新报告的艾滋病病毒（HIV）感染者接受抗病毒治疗前的CD4+ T淋巴细胞计数自然变化情况及其影响因素。方法 对2011年新报告且至少接受过2次CD4+ T淋巴细胞计数的HIV感染者进行分析,描述并比较其末次与首次CD4+ T淋巴细胞计数自然变化情况,运用Cox比例风险模型分析CD4+ T淋巴细胞计数显著下降的影响因素。结果 共有24 222例HIV感染者纳入分析,月均CD4+ T淋巴细胞计数变化速率中位数为-2.00（IQR:-7.81~3.601） cell/l,其中60.9%的HIV感染者末次CD4+ T淋巴细胞计数结果低于首次,且随着首末次检测间隔时间的延长,CD4+ T淋巴细胞计数下降明显。运用Cox比例风险模型分析发现,年龄较大和同性传播的HIV感染者CD4+ T淋巴细胞计数下降明显。结论 未经过抗病毒治疗的HIV感染者大部分仍处于疾病进展阶段,应加强对未接受抗病毒治疗的HIV感染者尤其是高年龄组以及同性传播感染者的随访,定期进行CD4+ T淋巴细胞计数检测,及早开展抗病毒治疗。     

Adult patients' adherence to anti-retroviral treatment: a survey correlating pharmacy refill records and pill counts with immunological and virological indices

Background

Providing anti-retroviral therapy (ART) to patients free of charge enhances their wellbeing, reduces the number of opportunistic infections and enables them to live productive lives, but only if these drugs are taken regularly every day for the rest of their lives. Patients’ ART adherence levels are difficult and expensive to estimate. If simple available measures, such as pharmacy refill records, could be correlated with laboratory test results for improved CD4 counts (indicating immunological recovery) and decreased viral loads (VLs – indicating virological recovery), these could be used as preliminary measures of adherence.

Objective

The objective was to determine whether the combination of pharmacy refill records and pill counts (including counts for all medications in the HAART regimen) could predict immunological and virologic responses through increased CD4 cell counts and suppressed VL.

Results

Correlations were compiled for patients’ pharmacy refill records, pill counts, CD4 cell counts, VL counts and self-reported ART adherence at one clinic in Gaborone, Botswana. There was a weak positive, relationship between ART adherence, CD4 cell counts and VL. Pharmacy refills and pill counts adherence measurement methodologies scored high on sensitivity and positive predictive values.

Conclusions

Pharmacy refill records and pill counts are useful indicators of patients’ ART adherence levels, but should be supplemented with CD4 and VL counts at regular intervals.     

多种炎性疾病患者外周血CD4＋和CD8＋T淋巴细胞的检测与临床意义分析

目的：探讨恶性肿瘤、肺结核、干燥综合征、葡萄膜炎等多种炎性疾病患者外周血CD4＋和CD8＋淋巴细胞的变化及其临床意义。方法应用BD FACSCanto II流式细胞仪检测上述疾病患者外周血CD4＋、CD8＋ T细胞并计算两者比值,同时检测健康者标本20例作为健康对照组,进行统计分析。结果与健康对照组比,恶性肿瘤组CD4＋ T淋巴细胞、CD4＋/CD8＋均明显降低,CD8＋ T淋巴细胞增高,差异有统计学意义（ P＜0．05）;全身炎性反应综合征组CD4＋ T淋巴细胞、CD4＋/CD8＋均明显降低,CD8＋ T 淋巴细胞增高,差异有统计学意义（ P＜0．05）;葡萄膜炎组CD8＋ T淋巴细胞增高,差异有统计学意义（P＜0．05）;肺结核组CD4＋ T淋巴细胞降低,差异有统计学意义（ P＜0．05）。结论患者的 CD4＋、CD8＋ T 淋巴细胞异常,表明机体细胞免疫功能异常。流式细胞仪检测CD4＋、CD8＋ T淋巴细胞可以作为患者临床免疫功能的初步诊断指标,辅助疾病治疗和病情观察。     

初治弥漫大B细胞淋巴瘤患者化疗后T淋巴细胞亚群变化

目的 探讨弥漫大B细胞淋巴瘤患者的细胞免疫状态改变及其预测预后的意义。方法 回顾性分析复旦大学附属中山医院厦门医院血液科2018年2月至2020年12月收治的28例初治弥漫大B细胞淋巴瘤患者。用流式细胞仪检测患者化疗前后不同时期外周血T淋巴细胞亚群变化。结果 与化疗前相比，患者化疗后外周血CD4⁺T淋巴细胞计数及CD4⁺/CD8⁺比值均降低。CD4⁺T淋巴细胞计数在化疗4个周期及6个周期后显著低于化疗前(P<0.05)，在化疗4个周期后达最低值[(0.245±0.086)×10⁹/L]。CD4⁺/CD8⁺比值在化疗4个周期后、化疗6个周期后、化疗结束后3个月及化疗结束后6个月显著低于化疗前(P<0.05)。终末疗效为完全缓解的患者化疗6个周期后CD4⁺T淋巴细胞计数及CD4⁺/CD8⁺比值高于非完全缓解患者(P<0.05)。复发患者化疗6个周期后CD4...     

SARS患者外周血CD4+CD8+T淋巴细胞的变化

目的 观察严重急性呼吸综合征(severe acute respiratory syndrome，SARS)患者外周血CD4^ 和CD8^ T淋巴细胞的变化，探讨SARS患者机体的免疫状况。方法 10位健康人(对照组)和13例确诊为SARS患者于发病第1、2、3、4周采静脉血，用流式细胞仪检测CD4^ 、CD8^ T淋巴细胞。结果 与对照组比较，SARS患者从发病1至4周外周血CD4^ 、CD8^ T淋巴细胞百分比均有不同程度的降低，以病程发展的2周左右为最明显。结论 SARS患者外周血CD4^ 、CD8^ T淋巴细胞有不同程度的降低，机体呈异常的免疫反应。     

Mutations to the probe of Cobas TaqMan HIV-1 ver. 1.0 assay causing undetectable viral load in a patient with acute HIV-1 infection

We encountered a human immunodeficiency virus (HIV)-1 in which the viral load was undetectable with the Cobas TaqMan HIV-1 ver. 1.0 (CTM v.1.0) in a patient with acute HIV-1 infection. The CTM v.1.0 assay showed more than 1,000-fold underestimation compared with the subsequent Cobas Amplicor Monitor v.1.5 assay. Because five mismatches to the CTM v.1.0 assay probe in the HIV-1 virus in the patient were disclosed by the manufacturer, partial gag regions of the HIV genome were directly sequenced from the patient’s plasma viral RNA. The detected single nucleotide point mutations were located near the 5′-end of the Cobas Amplicor Monitor probe. Clinicians should be very careful in making interpretations when indeterminate Western blot analysis results and a low or even undetectable HIV-1 viral load are encountered with the CTM HIV-1 ver. 1.0 assay in patients with suspected acute HIV infection. Repeating Western blot analysis is essential before considering a low HIV-1 viral load to be a false-positive result.     

柠檬酸碳酸氢盐透析液安全性评估及其对淋巴细胞亚群的影响

目的 研究柠檬酸碳酸氢盐透析液的安全性及其对维持性血液透析（MHD）患者外周血淋巴细胞亚群（CD4^＋、CD8^＋）和HLA-DR的影响。方法 选择西安交通大学第二医院肾内科30例2HD患者进行前后自身对照，前4周使用常规碳酸氢盐透析液（对照组），后4周使用柠檬酸酸碳酸氢盐透析液（实验组）。观察患者首次透析过程中生命体征。测定实验组首次透析前、透析后、透析后1h及第4周透析后1h血清离子钙（iCa^2＋）和柠檬酸的水平。两组首次透析前和第4周透析结束时分别测定CD4^＋、CD8^＋和HLA-DR表达率。结果 两组透析过程中生命体征平稳；实验组血清柠檬酸透析前后基本保持不变，iCa^2＋在透析后明显增高（P〈0．05）；两组透析后外周血淋巴细胞亚群（CD4^＋／CD8^＋比值）活性均增高，但对照组以CD4^＋升高为主，实验组以CD8^＋降低为主；HLA-DR在实验组透析后明显升高（P〈0．05），而对照组改变不明显。结论 柠檬酸碳酸氢盐透析液有其独特的优点，具有临床推广应用的价值。     

HIV-1 viraemia changes in patients with and without syncytium-inducing phenotype treated with nucleoside analogues: a case-control study

Abstract. The objective of the present study was to investigate the effect of the nucleoside analogue treatment on serum viraemia, CD4+ cell count and disease progression in patients with and without syncytium-inducing (SI) HIV-I variants. To achieve this in a case-control study, 11 nucleoside-naive patients harbouring SI variants who started treatment with zidovudine or zidovudine plus didanosine were matched with 11 control patients who never formed SI variants during a follow-up of 48 weeks. The matching criteria were age, CD4+ cell count and CDC clinical category at the start of the study and exposure to the same antiretroviral treatment. During the follow-up there were no significant differences in the changes of serum HIV-I RNA viral load and CD4+ cell counts between the two groups. In contrast, AIDS or new AIDS-defining events were observed in five SI cases but in none of the non-SI controls ( P = 0.002). The emergence of a zidovudine-resistant mutation at codon 215 was observed in all the patients harbouring SI strains and in six of the subjects with non-SI variants ( P = 0.03). The results of the present study show that in patients carrying SI virus, measurements of CD4+ count or RNA viral burden are neither related to the virulence of the virus strains nor able to predict the clinical course of the disease, at least under antiretroviral drug conditions. Thus, determination of SI phenotype should be considered in the evaluation and monitoring of HIV-I therapies.     

2008年河北省不同HIV-1亚型感染者的CD4＋／CD8＋T淋巴细胞检测分析

目的研究河北省新确认的不同亚型HIV-1感染者的CD4＋／CD8＋T淋巴细胞水平,探讨不同亚型人群的cD4＋／CD8＋T淋巴细胞水平有无差异。方法选取2008年1～12月新确认的HIV-1感染者55例,利用分子生物学方法进行分型后,对不同亚型人群的CD4＋／CD8＋检测数据进行分析统计。结果在55例已确定亚型的H1V-1感染者中,B亚型与CRF01-AE亚型、cRF07-BC亚型的CD4＋T淋巴细胞水平之间差异有统计学意义（P〈0．05）。CRF01-AE亚型、CRF07-BC亚型两者的CD4＋T淋巴细胞水平之间差异无统计学意义（P〉0．05）。但是B亚型与CRF01-AE亚型、CRF07-BC亚型三者之间的cD8＋T淋巴细胞水平差异无统计学意义（P〉0．05）。结论在新确认的3种不同亚型HIV-1感染者中,cD4＋T淋巴细胞水平是不同的,以B亚型感染者为最低;三者之间的cD8＋T淋巴细胞水平差异无统计学意义,推测可能与HIV-1感染途径和感染时间有关。     

rhG-CSF动员对异基因造血干细胞移植供者外周血CD4~+ T细胞S1P_1表达的影响

CD4+ T细胞主要通过其表面的S1P1受体与外界的第一信使1磷酸鞘氨醇(S1P)相互作用调节免疫功能。本研究旨在探讨重组人粒细胞集落刺激因子(rhG-CSF)动员对异基因造血干细胞移植(allo-HSCT)供者外周血CD4+ T细胞S1P1表达的影响。17例allo-HSCT供者于rhG-CSF动员前及动员后第4天采集外周血,使用磁珠分选法纯化CD4+ T细胞,提取微量RNA,用实时定量PCR法检测S1P1的表达。结果显示,rhG-CSF动员前后CD4+ T细胞均表达S1P1,且动员后S1P1在CD4+ T细胞中的表达明显低于动员前。结论:rhG-CSF动员使allo-HSCT供者外周血CD4+ T细胞S1P1的表达明显下调。     

调节性T淋巴细胞与反复体外受精-胚胎移植的研究

目的探讨外周血中调节性T淋巴细胞与反复体外受精-胚胎移植失败之间的相关性。方法用双荧光标记流式细胞分析技术,检测20例原因不明反复体外受精-胚胎移植失败患者、20例正常非孕妇女及18例第一周期体外受精-胚胎移植成功妇女外周血中CD4^＋CD25^＋调节性T淋巴细胞及CD4^＋CD25^highT细胞的水平。结果反复体外受精-胚胎移植失败患者外周血中CD4^＋CD25^＋调节性T淋巴细胞及CD4^＋CD25^highT细胞的百分率分别为（7．89±1．16）%和（1．06±0．33）％,低于正常非孕组的（11．73±1．05）％（P〈0．01）和（1．36±o．53）％（P〈0．05）,也低于第一周期体外受精-胚胎移植成功组的（9．67±1．87）％（P〈0．05）和（1．39±0．37）％（P〈0．05）。结论外周血中调节性T细胞的减少可能与反复体外受精-胚胎移植失败的发生有关。     

T细胞亚群在轻症/重症甲型H1N1流感患者中的差异及临床意义

目的 研究T淋巴细胞亚群在不同严重程度甲型H1N1流感患者中的差异及意义.方法 收集苏州大学附属第一医院2009年9月至2010年1月收治的66例经病毒检测确诊为甲型H1N1流感患者临床资料,按疾病严重程度及预后分为轻症治愈患者(B组,47例)、重症/危重症治愈患者(C组,14例)与死亡患者(D组,5例),以同期正常人群(A组,20例)作为对照组,回顾性比较各组患者在发病后不同时间点外周血淋巴细胞计数及T淋巴细胞亚群(CD3+,CD4+,CD8+)绝对值,检测方法采用流式细胞术;并比较各组患者的发热持续时间及病毒转阴时间.采用SAS9.13统计软件进行分析,多组间均数比较采用单因素方差及SNK法.结果 甲型H1 N1流感患者在发病初期即出现淋巴细胞计数以及T细胞亚群绝对值下降,与正常人群差异具有统计学意义(P＜0.05),B、C两组随病情好转逐步上升,C组淋巴细胞计数及T细胞亚群绝对值显著低于B组(P＜0.05),且C组恢复正常的时间晚于B组,而D组则持续偏低.三组患者发热时间/病毒转阴时间分别为(4.4±1.6)vs.(4.4±1.4)d、(12.9±3.1)vs.(10.2±2.6)d、(15.2±7.3)vs.(13.3±2.9)d,三组患者之间差异具有统计学意义(P＜0.05).结论 患者感染甲型H1N1流感病毒后,导致细胞免疫功能严重受损,且外周血淋巴细胞计数及T淋巴细胞亚群的变化与疾病严重程度及预后存在紧密联系.本研究结果可以用于指导临床诊断治疗.

Abstract：

Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P＜0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P ＜ 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P ＜ 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.     

367例HIV-1阳性样本WB确认结果的带型分析

目的通过对367例HIV-1抗体阳性者的蛋白印迹法（WB）的抗体条带分析,了解不同HIV-1感染人群其抗体WB带型分布的差异。方法采用WB法对HIV初筛有反应性样本进行HIV-1抗体确认,WB阳性者按照不同年龄、性别、民族三组进行WB带型的统计分析。结果 HIV-1gag基因编码的P39和P17抗体的阳性率在女性中明显高于男性（χ2值分别为6.386、10.872,均P〈0.05）;年龄小于或等于50岁的HIV-1抗体阳性者gag基因编码的P39和pol基因编码的P31抗体阳性率分别明显高于年龄超过50岁者（χ2值分别为6.882、4.360,均P〈0.05）;而HIV-1抗体WB带型分布在不同民族之间的差异无统计学意义。结论 HIV-1抗体WB带型分布在不同性别、不同年龄及不同民族人群间差异无统计学意义,HIV-1pol及gag基因编码的P39和P17蛋白的抗体产生在不同性别、不同年龄的人群间差异有统计学意义。这对HIV感染后病原检测和诊断包括传染源确定、传播途径追踪以及临床鉴别诊断具有非常重要的意义。     

Nef protein induces differential effects in CD8+ cells from HIV-1-infected patients

BACKGROUND: The Nef protein of HIV-1 is suspected to play a role in the depletion of uninfected CD4+ lymphocytes that leads to AIDS. By contrast its effect on CD8+ cells, whose functions are also deregulated during HIV-1 infection, is presently unclear. Here we describe a number of derangements induced in vitro by Nef in CD8+ cells from HIV-1-infected patients. DESIGN: Peripheral lymphocytes from 16 HIV-1+ subjects and 9 uninfected individuals were cultivated on a Nef-transfected mouse fibroblast layer exposing the carboxyl-terminal region of the viral protein on cell membrane. The cultures were then measured for both apoptosis and proliferation by subdiploid DNA content and Ki67 expression, respectively, whereas the molecular analysis of purified CD8+ cells investigated the Fas-L mRNA levels in Nef-treated CTLs. In addition, we evaluated the Nef-induced variation in the extent of CD8+/HLA-DR+ subset, which includes non cytotoxic cells secreting T-cell antiviral factor (CAF) and a soluble factor inhibiting the HIV-1 replication. RESULTS: The viral protein induced in peripheral blood lymphocytes (PBL) a moderate tendency to proliferate, as measured by the increment of Ki67 antigen, particularly on the CD8+ subset of HIV-1 infected individuals (P < 0.05). This profile was particularly evident in cultures from patients with severe CD4+ lymphopenia and paralleled an apparent expansion of the CD8+/CD57+ suppressor cell subset. Molecular analysis of purified CD8+ cells revealed a defective expression of Fas-L mRNA in Nef-cultured CTLs, whereas the viral protein exerted a down modulatory effect on the CD8+/HLA-DR+ subset (P < 0.05), thus suggesting a potential inhibition of CAF. CONCLUSIONS: These results support a potential role of Nef in the progression of HIV-1 infection as a number of cellular functions are affected in the CD8+ subset. In particular, the defective functions of CD8+ cells induced by the viral protein could contribute, at least partly, to the escape of HIV-1 from the immune control of these cells.