[慢性特发性血小板减少性紫癜中CD4(+) T细胞不同亚群的特征]

Chang Da-yu, Ouyang Jian, Zhou Rong-fu, Xu Jing-yan, Chen Bing, Yang Yong-gong, Zhang Qi-guo, Shao Xiao-yan, Guan Chao-yang, Xu Yong

Department of Hematology, The Affiliated Drum Tower Hospital, Nanjing University Medical School, Nanjing 210008, China.

Zhonghua Nei Ke Za Zhi. 2010 Mar;49(3):213-6.

OBJECTIVE

To explore the profiles of Th1, Th2, Th17 and Regulatory T (Treg) cells in patients with chronic idiopathic thrombocytopenic purpura (ITP).

METHODS

Samples of peripheral blood were collected from 35 chronic ITP patients (21 in an active stage group, 5 in a non-remission stage group, 9 in a remission stage group) and also from 18 healthy subjects. Flow cytometry was used to measure the intracellular cytokines interferon (IFN)gamma, interleukin (IL)-4 and IL-17 so as to identify the Th1, Th2 and IL-17 cells. Treg cells were identified with CD(4)(+) CD(25)(+) Foxp3(+) cells and uncultured peripheral blood was used to measure the CD(4)(+) CD(+)(25) Foxp3(+) cells with flow cytometry. The concentrations of IFNgamma, IL-4, IL-17 and IL-10 in plasma specimens were detected with ELISA method and its correlation with peripheral platelets counts and megakaryocytes number was analyzed, respectively.

RESULTS

There were no statistically significant differences between any two of the three groups for the percentage of Th1 cells, Th17 cells and Th1/Th17 ratio. The percentage of Th2 cells was (1.01 +/- 0.88)% in active stage and (1.22 +/- 1.04)% in non-remission stage, being significantly decreased than those in remission stage (1.93 +/- 1.04)% (P < 0.05) and the controls (1.86 +/- 0.59)% (P < 0.05). Th1/Th2 ratio was 15.04 +/- 9.67 in active stage, 11.65 +/- 9.32 in non-remission stage, which were significantly higher than those in remission stage (7.17 +/- 5.38, P < 0.05) and the controls (7.02 +/- 3.01, P < 0.05). The percentage of Treg cells was (0.89 +/- 0.58)% in active stage and (1.46 +/- 1.27)% in non-remission stage, being significantly decreased than those in remission stage (6.41 +/- 1.86)% (P < 0.01) and the control (5.73 +/- 0.71)% (P < 0.01). There was no statistic difference between any two of the three groups for plasma IFNgamma and IL-17 level. The plasma IL-4 level was (2.25 +/- 2.05) ng/L in active stage and (2.33 +/- 2.14) ng/L in non-remission stage, being significantly decreased than those in remission stage (6.00 +/- 4.57) ng/L (P < 0.05) and the controls (5.54 +/- 4.00) ng/L (P < 0.05). The plasma IL-10 level was (5.07 +/- 4.10) ng/L in active stage and (5.66 +/- 4.35) ng/L in non-remission stage, being significantly decreased than those in remission stage (10.92 +/- 6.17) ng/L (P < 0.01) and the controls (14.21 +/- 7.31) ng/L (P < 0.01). The plasma level of IL-10 in patients in active stage was positively related to the platelet counts (r = 0.16, P = 0.03).

CONCLUSION

Deficiency of Treg cells might be one of mechanisms that cause immune regulation dysfunction in chronic ITP. Th17 cells might not play a role in the development of chronic ITP.

目的

探讨慢性特发性血小板减少性紫癜（ITP）患者体内Th1、Th2、Th17及调节性T（Treg）细胞的特征。

方法

采集35例慢性ITP患者（活动期组21例、未缓解期组5例、缓解期组9例）及18例健康对照者的外周血样本。采用流式细胞术检测细胞内细胞因子干扰素（IFN）-γ、白细胞介素（IL）-4及IL-17，以鉴定Th1、Th2及IL-17细胞。通过CD4⁺CD25⁺Foxp3⁺细胞鉴定Treg细胞，采用流式细胞术检测未培养外周血中的CD4⁺CD25⁺Foxp3⁺细胞。采用ELISA法检测血浆标本中IFN-γ、IL-4、IL-17及IL-10的浓度，并分别分析其与外周血血小板计数及巨核细胞数量的相关性。

结果

活动期组、未缓解期组、缓解期组三组间Th1细胞、Th17细胞百分比及Th1/Th17比值差异均无统计学意义。活动期Th2细胞百分比为（1.01±0.88）%，未缓解期为（1.22±1.04）%，均显著低于缓解期（1.93±1.04）%（P<0.05）及健康对照组（1.86±0.59）%（P<0.05）。活动期Th1/Th2比值为15.04±9.67，未缓解期为11.65±9.32，均显著高于缓解期（7.17±5.38，P<0.05）及健康对照组（7.02±3.01，P<0.05）。活动期Treg细胞百分比为（0.89±0.58）%，未缓解期为（1.46±1.27）%，均显著低于缓解期（6.41±1.86）%（P<0.01）及健康对照组（5.73±0.71）%（P<0.01）。三组间血浆IFN-γ及IL-1水平差异无统计学意义。活动期血浆IL-4水平为（2.25±2.05）ng/L，未缓解期为（2.33±2.14）ng/L，均显著低于缓解期（6.00±4.57）ng/L（P<0.05）及健康对照组（5.54±4.00）ng/L（P<0.05）。活动期血浆IL-10水平为（5.07±4.10）ng/L，未缓解期为（5.66±4.35）ng/L，均显著低于缓解期（10.92±6.17）ng/L（P<0.01）及健康对照组（14.21±7.31）ng/L（P<0.01）。活动期患者血浆IL-10水平与血小板计数呈正相关（r=0.16，P=0.03）。