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多色流式细胞术 CD3 与 CD7 散点图反映了 HTLV-I 感染患者疾病阶段的进展。

The CD3 versus CD7 plot in multicolor flow cytometry reflects progression of disease stage in patients infected with HTLV-I.

机构信息

Division of Molecular Therapy, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

出版信息

PLoS One. 2013;8(1):e53728. doi: 10.1371/journal.pone.0053728. Epub 2013 Jan 22.

Abstract

PURPOSE

In a recent study to purify adult T-cell leukemia-lymphoma (ATL) cells from acute-type patients by flow cytometry, three subpopulations were observed in a CD3 versus CD7 plot (H: CD3(high)CD7(high); D: CD3(dim)CD7(dim); L: CD3(dim)CD7(low)). The majority of leukemia cells were enriched in the L subpopulation and the same clone was included in the D and L subpopulations, suggesting clonal evolution. In this study, we analyzed patients with indolent-type ATL and human T-cell leukemia virus type I (HTLV-I) asymptomatic carriers (ACs) to see whether the CD3 versus CD7 profile reflected progression in the properties of HTLV-I-infected cells.

EXPERIMENTAL DESIGN

Using peripheral blood mononuclear cells from patient samples, we performed multi-color flow cytometry. Cells that underwent fluorescence-activated cell sorting were subjected to molecular analyses, including inverse long PCR.

RESULTS

In the D(%) versus L(%) plot, patient data could largely be categorized into three groups (Group 1: AC; Group 2: smoldering- and chronic-type ATL; and Group 3: acute-type ATL). Some exceptions, however, were noted (e.g., ACs in Group 2). In the follow-up of some patients, clinical disease progression correlated well with the CD3 versus CD7 profile. In clonality analysis, we clearly detected a major clone in the D and L subpopulations in ATL cases and, intriguingly, in some ACs in Group 2.

CONCLUSION

We propose that the CD3 versus CD7 plot reflects progression of disease stage in patients infected with HTLV-I. The CD3 versus CD7 profile will be a new indicator, along with high proviral load, for HTLV-I ACs in forecasting disease progression.

摘要

目的

在最近一项通过流式细胞术从急性型患者中纯化成人 T 细胞白血病-淋巴瘤(ATL)细胞的研究中,在 CD3 与 CD7 的散点图中观察到三个亚群(H:CD3(高)CD7(高);D:CD3(低)CD7(低);L:CD3(低)CD7(低))。大多数白血病细胞富集在 L 亚群中,同一克隆包含在 D 和 L 亚群中,提示克隆进化。在这项研究中,我们分析了惰性型 ATL 患者和人类 T 细胞白血病病毒 I 型(HTLV-I)无症状携带者(AC),以观察 CD3 与 CD7 图谱是否反映了 HTLV-I 感染细胞特性的进展。

实验设计

使用来自患者样本的外周血单核细胞,我们进行了多色流式细胞术。对经荧光激活细胞分选的细胞进行分子分析,包括反转录长 PCR。

结果

在 D(%)与 L(%)的散点图中,患者数据可大致分为三组(组 1:AC;组 2:惰性和慢性型 ATL;组 3:急性型 ATL)。然而,也有一些例外(例如,组 2 中的 AC)。在一些患者的随访中,临床疾病进展与 CD3 与 CD7 图谱密切相关。在克隆性分析中,我们在 ATL 病例中清楚地检测到 D 和 L 亚群中的主要克隆,而且在组 2 中的一些 AC 中也存在,这令人好奇。

结论

我们提出 CD3 与 CD7 图谱反映了 HTLV-I 感染患者疾病阶段的进展。CD3 与 CD7 图谱将成为一个新的指标,与高前病毒载量一起,用于预测 HTLV-I AC 疾病进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b69/3551918/38b8e1c131e8/pone.0053728.g001.jpg

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