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采用羧基荧光素二乙酸琥珀酰亚胺酯法检测体外T淋巴细胞增殖,有助于原发性免疫缺陷病的诊断和管理。

In vitro T lymphocyte proliferation by carboxyfluorescein diacetate succinimidyl ester method is helpful in diagnosing and managing primary immunodeficiencies.

作者信息

Azarsiz Elif, Karaca Neslihan, Ergun Birgul, Durmuscan Mehmet, Kutukculer Necil, Aksu Guzide

机构信息

Department of Pediatric Immunology, Ege University, Izmir, Turkey.

出版信息

J Clin Lab Anal. 2018 Jan;32(1). doi: 10.1002/jcla.22216. Epub 2017 Apr 6.

Abstract

BACKGROUND

Functional studies besides routine laboratory tests for the definitive diagnosis of T lymphocyte disorders with isolated T or combined T/B-cell immunodeficiencies are important. We hereby summarized our experience with a carboxyfluorescein diacetate succinimidyl ester (CFSE)-based assay for the assessment of mitogenic T-cell proliferation responses in primary immunodeficiency (PID) patients who have not been diagnosed yet or genetically analyzed, but classified as probably having T-cell defects.

METHODS

Unclassified patients (n=46) and controls (n=25) were evaluated for T-cell disorders with CFSE-based assay.

RESULTS

CD3 blast cells after PHA-L stimulation were significantly lower in patients (31.1±28.8) than controls (67.9±8.79; P<.001). Nine patients with low and four patients with normal CD3 values had severely decreased blastic transformation. The proliferation response decreased mostly in combined immunodeficiency group. Sixteen of them had impaired proliferation responses. Appropriate molecular genetical analyses were planned after thorough evaluation of each patient.

CONCLUSIONS

In vitro lymphocyte cell proliferation analysis by CFSE method is a reliable and practical choice for the assessment of mitogenic T lymphocyte responses in yet unclassified PID patients for targeting further genetical analyses.

摘要

背景

除常规实验室检查外,功能研究对于明确诊断孤立性T淋巴细胞疾病或合并T/B细胞免疫缺陷的T淋巴细胞疾病至关重要。我们在此总结了我们使用基于羧基荧光素二醋酸琥珀酰亚胺酯(CFSE)的检测方法评估原发性免疫缺陷(PID)患者有丝分裂原T细胞增殖反应的经验,这些患者尚未被诊断或进行基因分析,但被归类为可能存在T细胞缺陷。

方法

使用基于CFSE的检测方法对未分类患者(n = 46)和对照组(n = 25)进行T细胞疾病评估。

结果

PHA-L刺激后的CD3母细胞在患者中(31.1±28.8)显著低于对照组(67.9±8.79;P<.001)。9例CD3值低的患者和4例CD3值正常的患者有严重的母细胞转化降低。增殖反应在联合免疫缺陷组中下降最为明显。其中16例患者的增殖反应受损。在对每位患者进行全面评估后计划进行适当的分子遗传学分析。

结论

通过CFSE方法进行体外淋巴细胞增殖分析是评估尚未分类的PID患者有丝分裂原T淋巴细胞反应以进行进一步基因分析的可靠且实用的选择。

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