Hunt Alicia M, Shallenberger Wendy, Ten Eyck Stephen P, Craig Fiona E
Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
Cytometry B Clin Cytom. 2016 Sep;90(5):404-14. doi: 10.1002/cyto.b.21335. Epub 2016 Mar 29.
Flow cytometry is an important tool for identification of neoplastic T-cells, but immunophenotypic abnormalities are often subtle and must be distinguished from nonneoplastic subsets. Use of internal control (IC) T-cells in the evaluation for T-cell neoplasms was explored, both as a quality measure and as a reference for evaluating abnormal antigen expression.
All peripheral blood specimens (3-month period), or those containing abnormal T-cells (29-month period), stained with CD45 V500, CD2 V450, CD3 PE-Cy7, CD7 PE, CD4 Per-CP-Cy5.5, CD8 APC-H7, CD56 APC, CD16&57 FITC, were evaluated. IC T-cells were identified (DIVA, BD Biosciences) and median fluorescence intensity (MFI) recorded. Selected files were merged and reference templates generated (Infinicyt, Cytognos).
IC T-cells were present in all specimens, including those with abnormal T-cells, but subsets were less well-represented. IC T-cell CD3 MFI differed between instruments (p = 0.0007) and subsets (p < 0.001), but not specimen categories, and served as a longitudinal process control. Merged files highlighted small unusual IC-T subsets: CD2+(dim) (0.25% total), CD2- (0.03% total). An IC reference template highlighted neoplastic T-cells, but was limited by staining variability (IC CD3 MFI reference samples different from test (p = 0.003)).
IC T-cells present in the majority of specimens can serve as positive and longitudinal process controls. Use of IC T-cells as an internal reference is limited by variable representation of subsets. Analysis of merged IC T-cells from previously analyzed patient samples can alert the interpreter to less-well-recognized non-neoplastic subsets. However, application of a merged file IC reference template was limited by staining variability. © 2016 Clinical Cytometry Society.
流式细胞术是鉴定肿瘤性T细胞的重要工具，但免疫表型异常往往很细微，必须与非肿瘤性亚群区分开来。探讨了在T细胞肿瘤评估中使用内部对照（IC）T细胞，既作为质量控制指标，也作为评估异常抗原表达的参考。
对所有外周血标本（3个月期间）或含有异常T细胞的标本（29个月期间）进行染色，染色抗体包括CD45 V500、CD2 V450、CD3 PE-Cy7、CD7 PE、CD4 Per-CP-Cy5.5、CD8 APC-H7、CD56 APC、CD16&57 FITC，并进行评估。通过DIVA（BD Biosciences）识别IC T细胞，并记录中位荧光强度（MFI）。合并选定的文件并生成参考模板（Infinicyt，Cytognos）。
IC T细胞存在于所有标本中，包括那些含有异常T细胞的标本，但亚群的代表性较差。IC T细胞CD3 MFI在不同仪器之间（p = 0.0007）和亚群之间（p < 0.001）存在差异，但在标本类别之间无差异，可作为纵向过程控制指标。合并后的文件突出显示了一些少见的IC-T亚群：CD2+(弱阳性)（占总数的0.25%）、CD2-（占总数的0.03%）。一个IC参考模板突出显示了肿瘤性T细胞，但受染色变异性的限制（IC CD3 MFI参考样本与测试样本不同，p = 0.003）。
大多数标本中存在的IC T细胞可作为阳性和纵向过程控制指标。将IC T细胞用作内部参考受到亚群代表性可变的限制。对先前分析的患者样本中的合并IC T细胞进行分析，可提醒解释人员注意那些不太被认可的非肿瘤性亚群。然而，合并文件IC参考模板的应用受到染色变异性的限制。©临床细胞计量学会2016年。
