[重型再生障碍性贫血中CD8(+)HLA-DR(+)效应T细胞调控因素的研究]

[Research of regulative factors on CD8(+)HLA-DR(+) effector T cells in severe aplastic anemia].

作者信息

Liu Xiao, Fu Rong, Wang Hua-quan, Liu Chun-yan, Ruan Er-bao, Qu Wen, Liang Yong, Wang Guo-jin, Wang Xiao-ming, Liu Hong, Wu Yu-hong, Song Jia, Xing Li-min, Guan Jing, Li Li-juan, Shao Zong-hong

机构信息

Department of Hematology & Oncology, General Hospital, Tianjin Medical University, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2012 Jun 26;92(24):1665-8.

PMID:22944154

Abstract

OBJECTIVE

To explore the regulative factors on CD8(+)HLA-DR(+) T cells in the patients with severe aplastic anemia (SAA) and examine the roles of these cells in the immunopathogenesis of SAA.

METHODS

CD8(+)HLA-DR(+) T cells were sorted from bone marrow mononuclear cells of 13 SAA patients from July 2011 to March 2012 by magnetic activated cell sorting system and were divided into 3 groups: interleukin 2 (IL-2) group (0, 0.1, 1, 10, 100 and 1000 U/ml), cyclosporine A (CsA) group (addition of 400 ng/ml CsA in each IL-2-containing well),receptor antagonist group (addition of IL-2 receptor antagonist 8 µg/ml in each IL-2-containing well). Then cell proliferation rate was evaluated by MTT assay after a 72-hour culturing. Bone marrow mononuclear cells of the SAA patients were divided into CsA group, IL-2 group and control group and cultured for 18 hours and another 4 hours following the dosing of phorbol ester. The expression of tumor necrosis factor β (TNF-β) in CD8(+)HLA-DR(+) T cells was analyzed by flow cytometry.

RESULTS

The cell proliferations of IL-2 wells at the concentrations of 10, 100 and 1000 U/L (0.36 ± 0.12, 0.41 ± 0.12, 0.46 ± 0.14) were significantly higher than those of the control wells (0.23 ± 0.11), CsA group (0.18 ± 0.05, 0.19 ± 0.00, 0.20 ± 0.04) and receptor antagonist group (0.18 ± 0.05, 0.17 ± 0.04, 0.18 ± 0.03, all P < 0.05). No statistic difference existed between CsA and receptor antagonist groups (P > 0.05). The expressions of TNF-β of CD8(+)HLA-DR(+)T cells of the IL-2 group were higher than those of the control group (64% ± 25% vs 46% ± 22%) whereas the CsA group (27% ± 20%) were lower than those of the control group (both P < 0.05).

CONCLUSIONS

IL-2 can significantly stimulate the proliferation of CD8(+)HLA-DR(+) T cells and accelerate the in vitro secretion of TNF-β in SAA patients. The proliferation may be inhibited by CsA and receptor antagonist. And the expression of TNF-β is suppressed significantly by CsA.

摘要

目的

探讨重型再生障碍性贫血（SAA）患者CD8(+)HLA-DR(+) T细胞的调节因素，并研究这些细胞在SAA免疫发病机制中的作用。

方法

采用磁性激活细胞分选系统从2011年7月至2012年3月期间13例SAA患者的骨髓单个核细胞中分离出CD8(+)HLA-DR(+) T细胞，并分为3组：白细胞介素2（IL-2）组（0、0.1、1、10、100和1000 U/ml）、环孢素A（CsA）组（在每个含IL-2的孔中加入400 ng/ml CsA）、受体拮抗剂组（在每个含IL-2的孔中加入8 μg/ml IL-2受体拮抗剂）。培养72小时后，采用MTT法评估细胞增殖率。将SAA患者的骨髓单个核细胞分为CsA组、IL-2组和对照组，培养18小时，加入佛波酯后再培养4小时。采用流式细胞术分析CD8(+)HLA-DR(+) T细胞中肿瘤坏死因子β（TNF-β）的表达。

结果

IL-2浓度为10、100和1000 U/L时孔中的细胞增殖率（0.36±0.12、0.41±0.12、0.46±0.14）显著高于对照组孔中的增殖率（0.23±0.11）、CsA组（0.18±0.05、0.19±0.00、0.20±0.04）和受体拮抗剂组（0.18±0.05、0.17±0.04、0.18±0.03，均P<0.05）。CsA组和受体拮抗剂组之间无统计学差异（P>0.05）。IL-2组CD8(+)HLA-DR(+)T细胞中TNF-β的表达高于对照组（64%±25%对46%±22%），而CsA组（27%±20%）低于对照组（均P<0.05）。