Milovanova Tatyana N
Department of Pathology and Laboratory Medicine, Flow Cytometry and Cell Sorting Facility, Abramson Cancer Center, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA.
Cytometry B Clin Cytom. 2007 Jul;72(4):265-75. doi: 10.1002/cyto.b.20166.
In this study, we evaluated alternative possibility for CFSE beryllium flow cytometric test against beryllium blood lymphocyte proliferation test (BeLPT) as a standard radioactive clinical screening method to identify sensitization to beryllium.
Delta PD (the ratio of divided cell population to the total number of cells with subtracted counts of unstimulated cells) of specific beryllium-induced pathogenic CD3+ CD4+ T-lymphocytes and stimulation index (SI) in CFSE proliferation test was compared with delta counts per minute (mean test CPM minus mean control CPM) and SI in radioactive blood BeLPT.
Comparison analysis of CFSE and BeLPT demonstrated excellent agreement between delta PD and delta CPM (kappa = 0.845, P << 0.0001). We determined 6.8% positive subjects in the beryllium-exposed, Be-LPT-negative group. The decreased mean difference of these indexes to percentage of average and the long tail in the plot reflects increased sensitivity. CFSE/CD4+ T-cell proliferation assay has 100% specificity, significantly higher sensitivity and efficiency than BeLPT.
Both delta PD, measured by the precursor frequencies method in CFSE assay and delta CPM, defined by tritiated thymidine in BeLPT, can be used for the enumeration of beryllium specific CD4+ T-cell proliferation and may substantially improve the quality of the early diagnosis of beryllium hypersensitivity.
在本研究中，我们评估了用羧基荧光素二醋酸盐琥珀酰亚胺酯（CFSE）铍流式细胞术检测替代铍血淋巴细胞增殖试验（BeLPT）作为标准放射性临床筛查方法来鉴定铍致敏的可能性。
将特定铍诱导的致病性CD3 + CD4 + T淋巴细胞的δPD（分裂细胞群体与扣除未刺激细胞计数后的细胞总数之比）和CFSE增殖试验中的刺激指数（SI）与放射性血液BeLPT中的每分钟δ计数（平均试验CPM减去平均对照CPM）和SI进行比较。
CFSE和BeLPT的比较分析表明，δPD和δCPM之间具有极好的一致性（kappa = 0.845，P << 0.0001）。我们在铍暴露、Be-LPT阴性组中确定了6.8%的阳性受试者。这些指标相对于平均值百分比的平均差异降低以及图中的长尾反映了敏感性增加。CFSE/CD4 + T细胞增殖试验具有100%的特异性，比BeLPT具有显著更高的敏感性和效率。
CFSE试验中通过前体频率法测量的δPD和BeLPT中由氚化胸腺嘧啶定义的δCPM均可用于计数铍特异性CD4 + T细胞增殖，并可能显著提高铍超敏反应早期诊断的质量。
