[慢性特发性血小板减少性紫癜中1型和2型T细胞的特征]

[Profiles of type 1 and type 2 T cells in chronic idiopathic thrombocytopenic purpura].

作者信息

Wang Ting-ting, Zhao Hui, Ren He, Guo Jian-hai, Xu Mao-qiang, Yang Ren-chi, Han Zhong-chao

机构信息

State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, CAMS and PUMC, Tianjin 300020, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2006 Mar 14;86(10):669-73.

PMID:16681922

Abstract

OBJECTIVE

To explore the profiles of type 1 and type 2 T cells in chronic idiopathic thrombocytopenic purpura (ITP) patients.

METHODS

Samples of peripheral blood were collected from 30 chronic ITP patients, 8 males and 22 females, aged 34, 20 being in the active stage, and 10 in the remission stage, and 20 healthy persons, 7 males and 13 females, aged 31. Peripheral blood mononuclear cells (PBMCs) were isolated, cultured, and activated with PMA/ionomycin. Flow cytometry was used to measure the intracellular cytokines interferon (IFN)-gamma and interleukin (IL)-4 in the PBMCs so as to identify the Th1 cells (IFN-gamma(+) IL-4(+) CD4(+) cells), Th2 cells (IFN-gamma(-) IL-4(+) CD4(+) cells), Tc1 cells (IFN-gamma+ IL-4(-) CD8(+) cells), and Tc2 cells (IFN-gamma(-) IL-4(+) CD8(+) cells). The ratios of Th2/Tc2, Th1/Th2, and Tc1/Tc2 were calculated. Samples of spleen tissue were collected from 8 patients with chronic ITP, 3 males and 5 females, aged 30, and 6 patients with hereditary spherocytosis (HS), 3 males and 3 females, aged 35, who underwent splenectomy. Splenocytes were isolated, cultured, and activated with PMA/ionomycin. Real-time PCR was used to detect the mRNA expression of T-bet and GATA-3 in the PBMCs and splenocytes.

RESULTS

The Th1/Th2 ratio of the patients in active stage was 25.62, significantly higher than those of the patients in remission stage (9.86) and the control (8.29, both P < 0.01), and the Tc1/Tc2 ratio of the patients in active stage was 30.23, significantly higher than those of the patients in remission stage (10.10) and the controls (12.58, both P < 0.01). The Th1/Th2 ratio of the splenocytes of the patients in active stage was 41.46, significantly higher than that of the controls (16.30, P < 0.01), and the Tc1/Tc2 ratio of the splenocytes of the active ITP patients was 35.80, not significantly higher than that of the controls (16.30, P = 0.082). The GATA-3 mRNA expression level of the PBMCs of the active ITP patients was one-fifth of that of the controls (P < 0.05) and the GATA-3 mRNA expression level of the splenocytes of the ITP patients was 0.34 of that of the HS patients (P < 0.05), however, there was no difference in the T-bet expression among the 3 groups. The T-bet/GATA-3 ratio of the PBMCs of the active ITP patients was 5.85 times that of the controls and the T-bet/GATA-3 ratio of the splenocytes of the active ITP patients was 2.68 times that of the controls (both P < 0.01).

CONCLUSION

ITP is a T1 cells (Th1 and Tc1) predominant disease. The T-bet/GATA-3 ratio may provide a surrogate marker of T1/T2 cytokine balance. Shifting the cytokine patterns from T1 to T2 may be a potential immunotherapy for ITP.

摘要

目的

探讨慢性特发性血小板减少性紫癜（ITP）患者1型和2型T细胞的特征。

方法

采集30例慢性ITP患者（男8例，女22例，年龄34岁，其中活动期20例，缓解期10例）及20例健康人（男7例，女13例，年龄31岁）的外周血样本。分离外周血单个核细胞（PBMC），进行培养，并用佛波酯/离子霉素激活。采用流式细胞术检测PBMC中细胞内细胞因子干扰素（IFN）-γ和白细胞介素（IL）-4，以鉴定Th1细胞（IFN-γ(+)IL-4(+)CD4(+)细胞）、Th2细胞（IFN-γ(-)IL-4(+)CD4(+)细胞）、Tc1细胞（IFN-γ+IL-4(-)CD8(+)细胞）和Tc2细胞（IFN-γ(-)IL-4(+)CD8(+)细胞）。计算Th2/Tc2、Th1/Th2和Tc1/Tc2的比值。采集8例慢性ITP患者（男3例，女5例，年龄30岁）及6例遗传性球形红细胞增多症（HS）患者（男3例，女3例，年龄35岁）行脾切除术后的脾组织样本。分离脾细胞，进行培养，并用佛波酯/离子霉素激活。采用实时定量PCR检测PBMC和脾细胞中T-bet和GATA-3的mRNA表达。

结果

活动期患者的Th1/Th2比值为25.62，显著高于缓解期患者（9.86）和对照组（8.29，P均<0.01）；活动期患者的Tc1/Tc2比值为30.23，显著高于缓解期患者（10.10）和对照组（12.58，P均<0.01）。活动期患者脾细胞的Th1/Th2比值为41.46，显著高于对照组（16.30，P<0.01）；活动期ITP患者脾细胞的Tc1/Tc2比值为35.80，显著高于对照组（16.30，P=0.082）。活动期ITP患者PBMC中GATA-3 mRNA表达水平为对照组的五分之一（P<0.05），ITP患者脾细胞中GATA-3 mRNA表达水平为HS患者的0.34（P<0.05），然而，3组间T-bet表达无差异。活动期ITP患者PBMC中T-bet/GATA-3比值为对照组的5.85倍，活动期ITP患者脾细胞中T-bet/GATA-3比值为对照组的2.68倍（P均<0.01）。