Beckman和BD两种机型流式细胞仪检测CD+4 T淋巴细胞数比较研究

目的 比较两种机型(Beckman Coulter FC500和BD FAC-S Calibur)流式细胞仪、两种"设门"方法平行检测艾滋病病毒(HIV)感染者CD 4 T淋巴细胞绝对数. 方法 用两种机型流式细胞仪、前向与侧向散射光和CD45两种"设门"方法,分3批次平行检测48份HIV感染者血标本. 结果 两种机型流式细胞仪、两种"设门"方法检测CD 4 T淋巴细胞计数结果差异无统计学意义(P＞0.05),并存在高度的相关性(P＜0.05),相关系数分别为0.941、0.966和0.970. 结论 两种机型、两种"设门"方法检测CD 4 T淋巴细胞计数结果接近,且高度相关,可以互为参考.     

HIV感染者CD_4^＋T淋巴细胞计数及血浆和尿液人巨细胞病毒检测的临床意义

目的分析艾滋病病毒（HIV）感染者的CD＋4T淋巴细胞（简称CD4细胞）计数,以及血浆和尿液中人巨细胞病毒（HCMV）脱氧核糖酸（DNA）检出率的差异性及其临床意义。方法 2012年7月至2013年7月间,对杭州市西溪医院住院的202例HIV感染者的血浆和尿液标本检测HCMV核酸,分析两种类型的标本HCMV-DNA检出率的差异性,同时观察CD4细胞计数与HCMV-DNA表达的关系并分析其临床意义。结果 202份血浆中HCMV-DNA阳性检出率为29.70%（60份）,202份尿液中HCMV-DNA检出率为47.03%（95份）,两种类型标本HCMV-DNA检出率差异有统计学意义（χ^2=5.79,P〈0.05）。不同CD4细胞计数的病人,其HCMV-DNA检出率也存在差异。在85例CD4细胞计数〉250个/μL组病人的尿液、血液中HCMV-DNA检出率分别为20.00%、2.35%;75例CD4细胞计数101-250个/μL组病人的尿液、血液中HCMV-DNA检出率分别为61.33%、44.00%;42例CD4细胞计数〈100个/μL组病人的尿液、血液中,HCMV-DNA检出率分别为76.19%、59.52%。随着CD4细胞计数的下降,尿液、血液中的HCMV-DNA检出率逐渐升高,各组间比较差异有统计学意义（χ^2=15.25,P〈0.05）。结论 HIV感染者血浆、尿液HCMV-DNA检出率存在差异,外周血CD4细胞计数越低越易发生HCMV感染。     

HIV感染者CD_4~+T淋巴细胞计数及血浆和尿液人巨细胞病毒检测的临床意义

目的分析艾滋病病毒(HIV)感染者的CD+4T淋巴细胞(简称CD4细胞)计数,以及血浆和尿液中人巨细胞病毒(HCMV)脱氧核糖酸(DNA)检出率的差异性及其临床意义。方法 2012年7月至2013年7月间,对杭州市西溪医院住院的202例HIV感染者的血浆和尿液标本检测HCMV核酸,分析两种类型的标本HCMV-DNA检出率的差异性,同时观察CD4细胞计数与HCMV-DNA表达的关系并分析其临床意义。结果 202份血浆中HCMV-DNA阳性检出率为29.70%(60份),202份尿液中HCMV-DNA检出率为47.03%(95份),两种类型标本HCMV-DNA检出率差异有统计学意义(χ2=5.79,P0.05)。不同CD4细胞计数的病人,其HCMV-DNA检出率也存在差异。在85例CD4细胞计数250个/μL组病人的尿液、血液中HCMV-DNA检出率分别为20.00%、2.35%;75例CD4细胞计数101~250个/μL组病人的尿液、血液中HCMV-DNA检出率分别为61.33%、44.00%;42例CD4细胞计数100个/μL组病人的尿液、血液中,HCMV-DNA检出率分别为76.19%、59.52%。随着CD4细胞计数的下降,尿液、血液中的HCMV-DNA检出率逐渐升高,各组间比较差异有统计学意义(χ2=15.25,P0.05)。结论 HIV感染者血浆、尿液HCMV-DNA检出率存在差异,外周血CD4细胞计数越低越易发生HCMV感染。     

北京市疾病预防控制中心VCT门诊点MSM的HIV检测和导医情况

目的评估2012年北京市疾病预防控制中心(CDC)艾滋病病毒(HIV)自愿咨询检测(VCT)门诊点,男男性行为人群(MSM)的HIV检测水平和HIV感染确诊后的导医情况。方法采用回顾性研究。收集2012年1月1日至12月31日北京市CDC VCT门诊点MSM的HIV检测和HIV感染确诊后导医情况等相关数据,并按月份整理。删除重复检测者和既往阳性者后,收集新确诊阳性者的CD+4T淋巴细胞(简称CD4细胞)检测和抗病毒治疗情况。利用Cox比例风险模型评估HIV感染确诊后CD4细胞水平和开始抗病毒治疗(ART)的相关性。结果2012年,北京市CDC VCT门诊总共检测HIV和梅毒1639人次,其中52.9%(867/1639)是MSM,142例为新发现的HIV感染者/AIDS病人。新发现的HIV感染者中,4.9%(7/142)是现患梅毒,12.7%(18/142)是既往梅毒患者。90.1%(128/142)的新发现HIV感染者已进行了阳性告知,85.2%(121/142)检测了CD4细胞计数,48.8%(59/121)的CD4细胞计数350个/μL。在121名接受了CD4细胞检测的MSM中,15名CD4细胞计数200个/μL的MSM开始了治疗;在40名CD4细胞计数为200~350个/μL的MSM中,有30人(75.0%)开始了治疗。结论 2012年,北京市MSM中新发现HIV感染的人数依然在增加,HIV阳性MSM开始抗病毒治疗的比例依然有待提高。     

三个省份中吸毒和有偿献血人群HIV感染者的现况调查

目的 了解吸毒者和有偿献血员中艾滋病病毒（HIV）感染者的现状、家庭内传播和抗病毒治疗开展的情况。方法 采用分层随机整群抽样法对3个省共163名HIV感染者进行问卷调查，调查数据经方差分析、卡方检验等统计方法处理。结果 不同传播途径HIV感染者的社会人口学指标年龄、性别明显不同。吸毒感染者平均年龄32．8岁，男性占83．9％；有偿献血员平均年龄43．5岁，男女比例接近。吸毒人群HIV感染者处于艾滋病（AIDS）阶段（CD4细胞计数〈200以1）的最多，占其总数的54．5％；有偿献血员人群中最低，占23．3％，有偿献血员中CD4细胞计数200～350/μl的最多（51．1％）。知道对方感染时夫妻生活每次都使用安全套的比率不高（58．3％）；18％（20／111）的HIV感染者配偶发生了夫妻间性传播，逻辑回归分析结果表明，与安全套使用情况显著相关；5．4％的HIV感染者的配偶至今没有检测是否感染HIV。不同传播途径HIV感染者的免费抗病毒治疗比例都在70％以上，有偿献血员治疗的比例最高。平均的免费治疗率90．5％，放弃治疗的平均比率为7．5％。结论 HIV的夫妻间性传播率很高，夫妻间安全套使用率低，要进一步加强宣传教育，提高预防意识。免费抗病毒治疗的比率很高，治疗的连续性较好。     

ELISA法检测尿液中HIV-1抗体的研究分析

目的 探讨艾滋病病毒(HIV)感染者晨尿、非晨尿和血液中HIV-1抗体检测结果的一致性，引进尿液HIV-1抗体检测方法。方法 平行采集吸毒人员血液和尿液标本，分别用血液和尿液酶联免疫吸附试验(ELISA)法检测HIV抗体，阳性血清标本送云南省疾病预防控制中心确认。结果 检测483人，血检阳性91人，晨尿检测阳性96人，两种方法一致性98．96％。对应晨尿阳性者采集非晨尿和阴性对照尿液标本各91份(其中血液标本检测HIV抗体阳性者86份)，检出阳性86份，阴性5份，非晨尿标本与血液标本检测结果一致。以血液标本检测结果为准，晨尿标本检测HIV-1抗体的灵敏度100％，特异度98．72％；非晨尿标本检测HIV-1抗体的灵敏度和特异度均为100％。结论 尿液ELISA法检测HIV-1抗体结果可靠，非晨尿可代替晨尿作HIV-1抗体筛查。     

2005年陕西省HIV耐药监测结果分析

目的 评估陕西省艾滋病高效抗逆转录病毒治疗（HAART）效果，及时发现艾滋病病毒（HIV）耐药毒株。方法 选择26例艾滋病（AIDS）治疗患者和2例待治疗HIV感染者作为监测对象，进行横断面调查，填写调查问卷，采集20ml抗凝血，检测病毒载量、CD4^＋T淋巴细胞计数和基因型耐药变异。结果 26例治疗患者在4种方案HAART治疗2～23个月（平均15个月）后，临床症状明显改善，20例（76．9％）治疗患者病毒复制得到有效控制，11例（42．3％）血浆病毒载量在检测水平以下；CD4^＋T淋巴细胞绝对计数平均为346．8±164．0个／mm^3，其中9例（34％）〉400个／mm^3。28例HIV／AIDS患者均未出现原发性耐药变异。结论 4种治疗方案疗效良好，均可有效抑制HIV-1复制，促进机体免疫重建，改善临床症状，无原发性耐药变异发生，未检出耐药毒株。     

2009-2012年湖州市艾滋病确证实验室HIV抗体检测结果分析

目的分析湖州市艾滋病网络实验室艾滋病病毒（HIV）抗体检测情况和艾滋病流行特征,为指导艾滋病防治提供科学依据。方法对HIV抗体初筛阳性标本,采用酶联免疫吸附试验（ELISA）和胶体硒法进行复检,两种试剂均呈阳性或一阴一阳的用蛋白免疫印迹法（WB）进行确证,确证阳性病例进行流行病学分析。结果717份HIV抗体初筛阳性标本,复核阳性333例,确证阳性321例,其中男性230例,女性91例。各类医院检查的占45．5％（146／321）;初中及以下文化占74．1％（238／321）;异性性传播占65．1％（209／321）,男男同性性传播占15．9％（51／321）。结论应加大健康教育和对高危人群行为干预的力度,遏制艾滋病的蔓延。     

PLG-CD4法测定HIV感染者CD4 T细胞

目的对美国Beckman-Coulter公司研发的测定艾滋病病毒(HIV)感染者CD4T细胞的PLG-CD4检测方法进行了测试,以验证该方法在检测新鲜血样及储存不同时间血样CD4T细胞所获的数据的准确性、稳定性和可靠性。方法对现场采集的血样室温放置,连续5天应用PLG-CD4法进行检测,其中第1天新鲜血样检测结果作为对照,并将结果进行平均值(Mean)、标准差(SD)和变异系数(CV)的统计计算。结果同一样品不同时间点检测的数据差异<20%为可接受范围。39份血液中有28份的5次重复测定值的CV值<10%,占72%。进一步分层显示:在CD4T细胞绝对计数<100时,易出现偏差(CV值为13%),而>100时,CV值均<10%。结论PLG-CD4技术简便易行,结果准确,重复性好,具有可检测陈旧血样CD4T细胞绝对计数的独到优势。     

昆明市HIV/AIDS监测结果分析

目的：及时掌握昆明市艾滋病病毒(HIV)的流行动态，特别是对一般人群的传播范围。方法：对监测人群的血清流行病学资料进行分析。结果：2001年共检测血清标本3528份，检出55例HIV感染者，检出率为1．56％。其中哨点监测1738人，检出36名HIV感染者；面上扩大监测1590人，检出6名HIV感染者；匿名检测200人，检出13名HIV感染者；检出率分别为2．07％、0．38％、6．50％。HIV感染者本市占74．55％，外地占25．45％。男性多于女性，其比例为2．05：1。年龄在16—39岁的占96．36％；职业以无业、农民为主。56．36％是通过静脉吸毒感染，43．64％通过性途径感染。结论：HIV进入一般人群的危险性增大；性传播有明显上升趋势，构成年为43．64％，与2000年(28．17％)相比差异有非常显著的统计学意义(P＜0．01)。控制吸毒特别是性途径造成的HIV传播，提高重点人群和一般人群的自我防范意识，是降低艾滋病危害的重要途径。     

HIV/AIDS病人CD_4^＋、CD_8^＋T淋巴细胞检测结果与标本存放时间的相关性研究

目的研究血样处理前后的存放时间,对艾滋病病毒（HIV）感染者/艾滋病（AIDS）病人（简称HIV/AIDS病人）CD_4^＋、CD_8^＋T淋巴细胞（简称CD4、CD8细胞）检测的影响。方法选取2013年新确证的HIV/AIDS病人119例,其抗凝全血室温放置6、24、48、72小时用FACSCalibur流式细胞仪分别检测CD4、CD8细胞绝对数,在样品处理后4℃放置0、24、48、72小时再上机检测CD4、CD8细胞绝对数;结果用EPI INFO软件进行统计学分析。结果抗凝全血存放24、48、72小时,与存放6小时相比,CD4、CD8细胞绝对数变化均无差异（P〉0.05）;血样处理后存放24、48、72小时,其CD4细胞数与存放0小时相比变化无差异（P〉0.05）,而血样处理后存放48小时,其CD8细胞数变化差异有统计学意义（P〈0.05）。结论抗凝全血在室温放置72小时,检测CD4、CD8细胞结果是可靠的,所以血样采集后放置时间可延长至72小时,处理过的血样在4℃放置72小时,虽然CD4细胞结果是可靠的,但CD8细胞却发生了明显变化,其二者比例也将随之发生变化,故处理后的样品应在24小时内完成检测。     

Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high on antiretroviral therapy

OBJECTIVE: To examine the relationships between blood CD4 natural regulatory T (Treg) cells, plasma HIV RNA level, CD4 T-cell count and immune activation in untreated HIV-infected patients and immunodeficient patients beginning antiretroviral therapy (ART), using a novel phenotype to define Treg cells (CD25CD127CD4). Data were compared with established Treg cell markers (FoxP3, CTLA-4 and GITR). METHODS: Twenty-nine untreated HIV-infected patients with CD4 T-cell counts of < 300 or > 400/microl were compared in a cross-sectional study and 12 patients beginning combination ART with < 100 CD4 T cells/mul were followed for 1 year on therapy. Three- and four-colour flow cytometry was used to quantitate proportions of Treg cells. RESULTS: In control donors and patients with high CD4 T-cell counts, 28-89% (median 60%) of CD25CD127CD4 cells were FoxP3, but < 10% expressed GITR or CTLA-4. Immunodeficient patients also had CD4-negative lymphocytes with the phenotype FoxP3CD127. Proportions of CD25CD127 cells and activated (HLA-DR) cells in the CD4 T-cell population were increased in patients with low CD4 T cell counts. The proportion of CD25CD127CD4 T cells correlated positively with plasma HIV RNA level and CD4 T-cell activation, but inversely with CD4 T-cell count. Longitudinal studies of 12 patients receiving ART in two distinct cohorts (Western Australia and Malaysia) showed that the proportion of CD25CD127CD4 cells decreased slightly over time, but remained above levels seen in non-HIV controls. CONCLUSIONS: Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high after 1 year on ART.     

Large diurnal variation in CD4 cell count and T-cell function among drug users: implications for clinical practice and epidemiological studies.

OBJECTIVE: To measure diurnal variation in the CD4 cell count and T-cell reactivity of drug users. DESIGN: A prospective epidemiological study among HIV-infected and non-infected drug users attending the Municipal Health Service of Amsterdam, The Netherlands. PATIENTS: Eleven HIV-infected and seven non-infected drug users. MAIN OUTCOME MEASURES: CD4 cell counts and T-cell reactivity three times a day. T-cell subsets and T-cell reactivity were determined from blinded samples within 2 hours. RESULTS: The number of CD4 cells increased by 130 x 10(6)/l (P < 0.05) in HIV-infected intravenous drug users over 8 hours. Following stimulation with anti-CD3 monoclonal antibodies, the T-cell reactivity of HIV-infected drug users rose from 118 to 221 c.p.m. (P < 0.01) over 8 hours. CD4 cell counts of the total study population increased by 37% and T-cell reactivity by 93%. The increase in the number of CD4 cells was more marked among active drug users than among drug users who had not used drugs recently. CONCLUSION: Variation in the CD4 cell count and in T-cell reactivity is large among drug users.     

Evaluation of a quantitative determination of CD4 and CD8 molecules as an alternative to CD4+ and CD8+ T lymphocyte counts in Africans

In the developed word, monitoring HIV-infected patients is routinely determined by CD4+ T lymphocyte absolute counts. The reference procedure, flow cytometry, is expensive, requires sophisticated instrumentation and operators with specific training. Due to these limitations, CD4 counting is often unavailable in developing countries. The Capcellia assay is an enzyme-linked immunoassay for quantitative determination of CD4 and CD8 molecules. We evaluated this method in West Africa on blood samples collected from 39 HIV-uninfected and 44 HIV-infected adult subjects. CD4 concentration ranges were determined according to the clinical stages of the disease. We then studied the relationship between the two methods in the HIV-infected patients. The Spearman's rank correlation was 0.61 (95% confidence interval: 0.38-0.76, P < 0.0001). Nevertheless, determination of limits of agreement revealed discrepancies between the two methods, especially for CD4 counts > 0.4 x 10(9)/l, which are discussed. We conclude that the Capcellia assay is a convenient means to determine the immunodepression level where flow cytometric instrumentation is unavailable, and can be complementary to CD4 T lymphocyte enumeration.     

Successful implementation of a low-cost method for enumerating CD4+ T lymphocytes in resource-limited settings: the ANRS 12-26 study

OBJECTIVE: To evaluate the feasibility and the relevance of the implementation of an alternative technique to flow cytometry (FC) for enumerating CD4 T cells (Dynabeads; Dynal Biotech, Oslo, Norway), based on quantifying CD4 T cells by epifluorescent microscopy following their isolation using anti-CD4 monoclonal antibody-coated magnetic beads. DESIGN: International multi-center study. Five consecutive runs of dual CD4 T-lymphocyte enumeration by both techniques in six sites in five countries of West Africa. METHODS: A total of 657 pairs of values of CD4 cell counts were generated by 43 technicians by both FC (TruCount; Becton Dickinson Immunocytometry Systems, San Jose, California, USA) and Dynabeads from blood samples obtained from 301 HIV-infected patients, seen in one (n = 112), two (n = 61), three (n = 75), four (n = 40) or five (n = 13) occasions. RESULTS: The correlation coefficient between the results of the two techniques was 0.89. The overall systematic difference between Dynabeads and FC was -16 x 10(6) cells/l (P < 10(-4)). The median difference was insignificant (+7.5 cells) for CD4 cell counts below 200 x 10(6) cells/l and increased with CD4 levels. Patients were consistently classified at the threshold of 200 x 106 cells/l by both methods in 88.7% of cases. Among the 74 discrepant pairs of values, only 31 (4.7%) exhibited a difference of more than 100 x 10(6) cells/l. CONCLUSIONS: Results from Dynabeads and FC were highly correlated. The ability of the alternative method to consistently classify results in agreement with FC, at thresholds of CD4 cell counts relevant for clinical care, was high. The implementation of this low-cost method was easy and successful in the West African context.     

Rate of decline of absolute number and percentage of CD4 T lymphocytes among HIV-1-infected adults in Dar es Salaam, Tanzania

OBJECTIVE: To determine the rate of decline of CD4 T lymphocytes among HIV-1-infected individuals. DESIGN AND SETTING: A prospective open cohort study of workers in three hotels in Dar es Salaam. METHODS: The workers were seen yearly during the study. CD4 T lymphocyte counts were determined using flow cytometry. The CD4 T-lymphocyte slopes were determined using a linear regression model. RESULTS: During the 9-year study period 682 subjects were selected for lymphocyte subset determinations. Of these, 94 HIV-1-seroprevalent (72%), 77 HIV-1-seroincident (67%) and 325 seronegative (75%) individuals had three or more CD4 T-cell determinations, and were used for calculations of CD4 cell slopes with a mean follow-up period of 71.4, 52.9 and 86.0 months, respectively. The median yearly decline of the CD4 T-lymphocyte counts and percentages among seroprevalent individuals was -21.5 cells/microl and -1.3%; among the seroincident individuals the median decline was -22.0 cells/microl and -1.5%. In seroincident individuals the mean duration to a CD4 T-lymphocyte level corresponding to a definition of AIDS was 13.3 years or 11.8 years for CD4 cell counts or percentages, respectively. HIV-1-seropositive subjects who died had significantly steeper CD4 cell slopes than those who survived. CONCLUSION: The rates of CD4 T-lymphocyte decline in HIV-1-infected individuals in our population are similar to those reported in Europe and north America.     

HIV/AIDS患者中CD4细胞计数与总淋巴细胞计数间相关性研究

目的评价总淋巴细胞计数与CD4细胞计数间的相关性.方法回顾性分析了226例艾滋病病毒(HIV)阳性患者共330对同一天获得的CD4细胞计数与总淋巴细胞计数间的相关性,阳性预测值(PPV)、敏感性、特异性分别在不同的总淋巴细胞计数范围对应于CD4细胞计数<200个/mm3和CD4细胞计数<350个/mm3时获得.结果 330对CD4细胞计数与总淋巴细胞计数之间存在相关性(r=0.528,P<0.01),总淋巴细胞计数<1 400个/mm3对应于CD4细胞计数<200个/mm3有70.11%的阳性预测值,72.61%的敏感性,88.46%的特异性,总淋巴细胞计数<1 900个/mm3对应于CD4细胞计数<350个/mm3有80.97%的阳性预测值,70.92%的敏感性,74.54%的特异性.结论总淋巴细胞计数可以作为评价患者患机会性感染的危险程度及何时开始药物治疗的一种低消费的监测手段.阳性预测值(PPV)、敏感性、特异性分别在总淋巴细胞计数<1 400个/mm3对应于CD4细胞计数<200个/mm3和总淋巴细胞计数<1 900个/mm3对应于CD4细胞计数<350个/mm3时表现最为明显.     

Evaluation of the World Health Organization staging system for HIV infection and disease in Ethiopia: association between clinical stages and laboratory markers

OBJECTIVE: To study the association between the clinical axis of the World Health Organization (WHO) staging system of HIV infection and disease and laboratory markers in HIV-infected Ethiopians. DESIGN: Cross-sectional study. METHODS: Clinical manifestations and stage of HIV-positive individuals participating in a cohort study of HIV infection progression, and of HIV-positive patients hospitalized with suspicion of AIDS, were compared to CD4+ T-cell count and viral load. RESULTS: Of the 86 HIV-positive participants of the cohort study, 53 (62%), 16 (19%), 16 (19%), and one (1.2%) were in stage 1, 2, 3 and 4, respectively. Minor weight loss (n = 15) and pulmonary tuberculosis (n = 9) were the most commonly diagnosed conditions among the 38 (44%) symptomatic HIV-positive individuals. Although 23 (27%) HIV-positive participants had CD4+ T-cell counts less than 200 x 10(6)/l, only one was in clinical stage 4. Among 79 hospitalized HIV-positive patients, 15 (19%) and 64 (81%) were in stage 3 and 4, respectively. The majority (83.5%) had CD4+ T-cell counts < 200 x 10(6)/l. Individuals at stage 3 had lower CD4+ T-cell counts and higher viral loads when seen in hospital as compared to cohort participants (P = 0.06 and 0.008, respectively). When grouping the two study populations, the median CD4+ T-cell count decreased (337, 262, 225, 126, and 78 x 10(6)/l, P< 0.01), and the median viral load increased (4.08, 3.89, 4.47, 5.65, and 5.65 log10 copies/ml, P < 0.01), with increasing clinical stage of HIV infection (1, 2, 3 cohort, 3 hospital, and 4, respectively). Median CD4+ T-cell counts were remarkably low in HIV-negative participants (749 x 10(6)/l), and in HIV-positive participants at stage 1 and 2 (337 and 262 x 10(6)/l, respectively). CONCLUSIONS: There was a good correlation between WHO clinical stages and biological markers. CD4+ T-cell counts were low in Ethiopians, particularly during early stages of HIV-1 infection, and preliminary reference values at different stages of HIV-1 infection were determined. In HIV-infected Ethiopians, lymphocyte counts less than 1,000 x 10(6)/l in non-hospitalized individuals, and less than 2,000 x 10(6)/l in hospitalized patients, had high positive predictive value, but low sensitivity, in identifying subjects with low CD4+ T-cell counts (< 200 x 10(6)/l) who would benefit from chemoprophylaxis of opportunistic infections. The on-going longitudinal study will be useful to confirm the prognostic value of the WHO staging system.     

Dendritic cells generated in the presence of granulocyte-macrophage colony-stimulating factor and IFN-alpha are potent inducers of HIV-specific CD8 T cells

OBJECTIVE: To investigate the ability of granulocyte-macrophage colony-stimulating factor (GM-CSF) and IFN-alpha to induce the differentiation of peripheral monocytes into dendritic cells (DC) and their ability to trigger an HIV-specific CD8 T-cell response. METHODS: Monocytes isolated from both seronegative controls and HIV-infected individuals were differentiated into DC using GM-CSF with either IL-4 or IFN-alpha for 7 days. We assessed the phenotypic characteristics and IL-12 production by flow cytometry. The ability of DC to trigger CD8 T-cell responses was assessed by means of ELISpot and cytotoxicity assays. In addition, HIV-1-RNA levels were measured in culture supernatants. RESULTS: Compared with control DC generated in the presence of GM-CSF and IL-4, DC generated in the presence of GM-CSF and IFN-alpha expressed higher levels of MHC class I molecules and produced similar or higher levels of IL-12 after CD40 ligation or Staphyloccus aureus Cowan stimulation. GM-CSF/IFN-alpha DC expressed low levels of CD4, CXCR4 and DC-SIGN and did not produce detectable virus during the differentiation period. Pulsed GM-CSF/IFN-alpha DC were found to prime CD8 T cells from HIV-negative controls to exert cytotoxic activity against target cells expressing HIV antigens. HIV peptide-pulsed GM-CSF/IFN-alpha DC promote specific IFN-gamma production by autologous CD8 T cells from HIV-seronegative donors. Furthermore, GM-CSF/IFN-alpha DC from HIV-seropositive patients efficiently present HIV peptides to autologous CD8 T lymphocytes. CONCLUSION: GM-CSF and IFN-alpha allow the generation of DC with high CD8 T-cell stimulating abilities. Therefore, this strategy may represent a novel approach to therapeutic vaccination in HIV disease.