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.     

单平台TruCount和PLG-CD4测定CD4T细胞绝对计数的比较

目的了解单平台TruCount和PLG-CD4两种不同流式设门方法,对同一样品测定CD4T细胞绝对计数的影响。方法采集408份样品,包括同批次114份,不同时间15批次294份,比较两种不同设门方法检测同一样品CD4T细胞的绝对计数。结果对TruCount和PLG-CD4两种流式设门方法所获取的408份样品的CD4T细胞绝对计数进行分层后显示:只有4例在分层时落入不同范围,判定差异率<1%;在≤200个细胞/μl的范围内,PLG-CD4判定90例,比TruCount少判1例。两种不同方法所得CD4T细胞绝对计数差异均<10%,相关系数r=0.99。结论单平台TruCount和PLG-CD4两种不同流式设门方法测定的CD4T细胞绝对计数是一致的,实际应用中所采用的方法将决定于不同试剂的价格与操作的简易性。     

湖北成人CD4、CD8T淋巴细胞计数正常值调查及CD4与总淋巴细胞数的相关性分析

目的了解中国湖北成人CD4抗原阳性的T淋巴细胞(CD4)、CD8抗原阳性的T淋巴细胞(CD8)、CD3抗原阳性的T淋巴细胞(CD3)及CD4/CD8、CD4/CD3正常值参考范围,以及总淋巴细胞计数(TLC)与CD4之间的关系.方法选取120例15～60岁健康人不同年龄组的血标本,用Becton Dickinson公司生产的FACSCount自动化仪检测CD4、CD8、CD3、CD4/CD8、CD4/CD3,计算相关数据的正常值参考范围,并将其与国外数据作比较.用美国雅培CEILDYN(R)3700血液全自动分析仪作全血总淋巴细胞计数.结果CD4值为681±21/mm3(-x±Sd,以下相同),CD8值为510±26/mm3,CD3值为1 299±38/mm3,CD4/CD8值为1.43±0.06,CD4/CD3值为0.54±0.10,TLC值为1 752±48/mm3.CD4、CD8、CD3计数及CD4/CD8、CD4/CD3在不同性别、年龄组之间差别无显著性.CD4与TLC之间有显著的直线正相关性,r=0.647,回归方程为√CD4=0.422√TLC+8.444.结论CD4、CD3、CD8等正常值在不同性别、年龄间相近.中国湖北成人CD4值与中国上海成人之间无差别,但显著低于国外值.在无条件检测CD4计数的情况下,可用TLC推测CD4的值.中国现有的治疗标准需要进一步探讨.     

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。     

上海地区成人不同年龄组间CD4、CD8 淋巴细胞计数正常值调查

目的　了解上海地区成人CD4、CD8计数及CD4/CD8比值正常参考值范围。方法　选取 6 14例成年人不同年龄组的血标本 ,根据不同性别、不同年龄组分别进行比较 ,用流式细胞仪进行CD4、CD8计数检测 ,得出不同性别及不同年龄组的CD4、CD8计数及CD4/CD8比值 ,并进行分析。结果　CD4正常值在不同性别及年龄组之间差异无显著性 ,其正常平均值为 72 6 .99± 2 5 5 .2 1。CD8及CD4/CD8在不同性别及其年龄组之间差异有显著性。CD8计数平均值为 5 39.98± 134 .0 7。CD4/CD8平均值为 1.49± 0 .5 7。结论　上海地区成人CD4淋巴细胞计数较国外低 10 0 /mm3 左右。对获得性免疫缺陷综合征CD4平均值的诊断临界值建议可采用 40 0 /mm3 。     

2．CD4＋细胞计数急速下降

最近，我们收了一例HIV感染病人，她正在接受抗逆转录病毒治疗。2003年十二月，她最后的CD4 细胞计数为1063 cells／mcL，检测不到病毒负荷。2004年二月她停止了药物治疗而未进行随访。最近，她因肺炎住院，证明是卡氏肺囊虫，她现在的CD4 细胞计数仅为50 cells／mcL。CD4 细胞计数有可能下降得这么快吗?如果有可能的话，这种情况是常见的吗?对于这样的急速下降潜在的危险因素是什么?     

用于监测HIV感染者CD4细胞计数的新方法

泰国的研究人员表示，就监测HIV感染者CD4^＋细胞计数而言，一种新的、低成本的CD4筛选法可以替代现有复杂而又昂贵的流式细胞术的方法。新方法所产生的显而易见的临床意义在于可以使泰国和其他发展中国家的HIV感染者无需太多的花费就能够获得CD4^＋细胞计数的监测。     

HIV感染者CD4细胞计数与肺结核患病情况分析

河南省对部分艾滋病病毒（HIV）感染者进行了肺结核病筛查，并进行了HIV感染者CD。细胞计数与肺结核患病相关性分析。     

Affordable CD4(+) T-cell counts on 'single-platform' flow cytometers I. Primary CD4 gating

Here, we demonstrate the flow cytometric concept of 'primary CD4 gating' utilizing three different CD4 monoclonal antibodies (mAbs) conjugated with five different fluorochromes. CD4(+) lymphocytes were defined by an autogate in a single histogram of CD4 fluorescence intensity (FI) (y-axis) vs. side light scatter (x-axis). A wide range of absolute counts for > 600 individuals, including HIV(+) patients, were compared with those obtained by 'state-of-the-art' single-platform flow cytometers such as the volumetric Ortho CytoronAbsolute and the Becton Dickinson FACSCalibur using TruCount beads. The correlation between CD4 counts obtained with primary CD4 gating and the full test panel on the Ortho Cytoron was excellent (R(2) = 0.999). Bland-Altman statistics showed a mean difference of -2 cells/mm(3) [confidence interval (CI) 95% = -3 to -1; limits of agreement -27 to +23]. In addition to absolute CD4 counts, CD4% values and CD4/CD8 ratios are also frequently requested. To obtain these, lymphocytes need to be counted using scatter gates, and a second tube stained with a CD8 mAb to count CD8(++) lymphocytes can be incorporated. We conclude that primary CD4 gating on single-platform volumetric flow cytometers is one of the most economical and flexible technologies for routine cost-conscious service work, particularly during the follow-up of patients undergoing anti-HIV therapy and/or vaccination in the developing world.     

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.     

CD4 percentage is an independent predictor of survival in patients starting antiretroviral therapy with absolute CD4 cell counts between 200 and 350 cells/microL

Objective

To determine the prognostic value of baseline CD4 percentage in terms of patient survival in comparison to absolute CD4 cell counts for HIV‐positive patients initiating highly active antiretroviral therapy (HAART).

Methods

A population‐based cohort study of 1623 antiretroviral therapy‐naïve HIV‐positive individuals who initiated HAART between 1 August 1996 and 30 June 2002 was conducted. Cumulative mortality rates were estimated using Kaplan–Meier methods. Cox proportional hazards regression was used to model the effect of baseline CD4 strata and CD4 percentage strata and other prognostic variables on survival. A subgroup analysis was conducted on 417 AIDS‐free subjects with baseline CD4 counts between 200 and 350 cells/μL.

Results

In multivariate models, low CD4 percentages were associated with increased risk of death [CD4%<5, relative hazard (RH)=4.46; CD4% 5–14, RH=2.43; P<0.01 for both] when compared with those subjects with an initial CD4 fraction of 15% or greater, but had less predictive value than absolute CD4 counts. In subgroup analyses where absolute CD4 strata were not associated with mortality, a baseline CD4 fraction below 15% [RH=2.71; 95% confidence interval (CI) 1.20–6.10], poor adherence to therapy and baseline viral load >100 000 HIV‐1 RNA copies/mL were associated with an increased risk of death.

Conclusion

CD4 percentages below 15% are independent predictors of mortality in AIDS‐free patients starting HAART, including those with CD4 counts between 200 and 350 cells/μL. CD4 percentage should be considered for inclusion in guidelines used to determine when to start therapy.

     

Association between serum amylase levels and CD4 cell counts in newly diagnosed people living with HIV: A case-control study

Serum amylase is a direct reflection of pancreatic injury. Several clinical studies have indicated that antiretroviral therapy may be the main cause of increased serum amylase in people living with human immunodeficiency virus (PLWH). However, other probable causes including direct human immunodeficiency virus infection, opportunistic infections and neoplasms, alcohol abuse, and use of illicit drugs, which can also affect pancreatic amylase levels were not considered in these studies. In our study, we collected clinical data from newly diagnosed PLWH who had not received antiretroviral therapy, and examined the association between serum amylase levels and CD4 cell counts. Between November 2018 and September 2021, a total of 344 newly diagnosed PLWH and 344 healthy controls were recruited at Ningbo Yinzhou No 2 Hospital. Serum amylase levels, CD4 cell counts and other clinical features were measured. Relationships between serum amylase levels and clinical parameters were evaluated using correlation analysis. Multiple linear regression analyses were performed to identify the independent risk factors. Newly diagnosed PLWH had lower CD4 cell counts and higher serum amylase levels than healthy controls (P < .05). Serum amylase levels were negatively correlated with CD4 cell counts (r = −0.506, P < .001). In multiple linear regression analyses, CD4 cell counts (β = −0.327, 95% confidence interval = −0.051–−0.022, P < .001) were independently associated with serum amylase levels. CD4 cell counts were independently associated with serum amylase levels in newly diagnosed PLWH. Thus, close monitoring of serum amylase may be significant in preventing opportunistic infections of PLWH, since low CD4 cell counts are associated with an increased risk of opportunistic infections.     

HIV/AIDS患者外周血CD4+ T细胞计数和淋巴细胞计数相关性的多中心研究

目的 对淋巴细胞计数预测CD4+ T细胞计数的准确性进行评价,为临床应用提供支持.方法 在全国23个分中心共筛查2 013例未接受抗病毒治疗的HIV/AIDS患者,经流式细胞术检测CD4+ T细胞计数,分析血常规当中的淋巴细胞计数和CD4+ T细胞计数之间的相关性,绘制ROC曲线判断淋巴细胞计数预测CD4+T细胞计数的准确性,并计算其敏感度、特异度、阳性预测值和阴性预测值.结果 2 013例HIV/AIDS患者的淋巴细胞计数和CD4+ T细胞计数分别为(1 600±670)×106/L和(244±148)×106/L,两者呈现显著正相关性(r=0.482,P＜0.000 1),以淋巴细胞计数预测CD4T细胞计数＜100×106/L、＜200×106/L和＜350×106/L的AUCROC分别为0.790(95％ CI0.761 ～0.818,P＜0.000 1),0.733(95％ CI0.710 ～0.755,P ＜0.000 1)和0.732 (95％ CI0.706 ～0.758,P ＜0.000 1).结论 在HIV/AIDS患者的临床诊治中,用淋巴细胞计数预测CD4+ T细胞计数有其实用价值,可以考虑在不具备CD4+ T细胞计数直接检测时作为替代指标用于监测疾病进展,也可以在获得CD4+ T细胞计数结果之前用来初步快速判断患者病情.     

Comparison of different methodologies for CD4 estimation in a clinical setting

OBJECTIVE: To compare flow cytometry assays, using traditional dual platform (DP) or newer single platform (SP) for CD4 enumeration. METHOD: Records of subjects enrolled in four separate clinical trials using the same central laboratory [SP methodology (Trucount)] were reviewed retrospectively. Eighteen subjects had 60 matching contemporaneous samples at multiple timepoints. RESULTS: DP flow cytometry yielded higher CD4 counts in 50/60 assays (83%). CD4 count and percentage by the two methods showed strong correlation for the counts (r=0.965, P<0.0001) and percentages (r=0.959, P<0.0001). Bland-Altman plot analysis showed that the limits of variation were within agreeable limits of +/-2SD in 56/60 (93.3%) samples tested. Twenty-five (42%) samples had a difference of >50 cells/microL. Of these six (24%) exceeded 100 cells. CONCLUSION: This study is in agreement with previous reports of strong correlation between DP and SP flow cytometry. This review found differences in CD4 counts in a high proportion of samples tested highlighting the importance for clinicians to be aware of such differences when interpreting results from the two methods.     

Absolute CD4+ T-lymphocyte and CD34+ stem cell counts by single-platform flow cytometry: the way forward

To determine the potential advantage of single-platform technology in the enumeration of CD4+ T lymphocyte and CD34+ stem cells, data has been analysed from the UK NEQAS for Leucocyte Immunophenotyping schemes. The inter-laboratory CVs for CD4+ T lymphocyte counts were consistently lower for single-platform (mean 13.7%, range 10-18.3%) compared to dual-platform methodology (mean 23.4%, range 14.5-43.7%). Subgroup analysis of single-platform users demonstrated mean overall inter-laboratory CVs of 17.2%, 13% and 7.1% for the FlowCount, TruCount and volumetric approach respectively. The lowest inter-laboratory CVs obtained for a single sample by each single platform approach were 4% (TruCount), 4.4% (volumetric), 4.6% (FACSCount) and 12.7% (FlowCount). Similarly, the mean inter-laboratory CV for CD34+ stem cell enumeration using non-standardized single-platform approaches was 18.6% (range 3.1-36.9%) compared to 28.6% (range 19-44.2%) for the dual-platform technology. Our results suggest absolute cell subset enumeration should be performed by single-platform technology and that such an approach should improve the quality control of multi-centre clinical trial data for CD4+ T lymphocyte and CD34+ stem cells.