CD8CD28/CD8CD28 T淋巴细胞平衡失调与三硝基苯磺酸诱导的大鼠结肠炎预后相关。

Disequilibrium in the CD8CD28/CD8CD28 T Lymphocyte Balance Is Related to Prognosis in Rats with Trinitrobenzenesulfonic Acid-Induced Colitis.

作者信息

Dai Shixue, Gu Hongxiang, Lin Qianyi, Xing Tiaosi, Chen Minhua, Zhong Tao, Wu Gang, Feng Yanling, Liu Hongbo, Gao Yong, Jian Hongjian, Zhang Minhai, Mo Hongmei, Zhu Huanjie, Chen Dongsheng, Xu Jun, Zou Ying, Chi Honggang, Zhu Yuzhen

机构信息

Department of Rheumatology, TCM-Integrated Hospital, Southern Medical University, No. 13, Shiliugang Road, Haizhu District, Guangzhou, 510315, Guangdong, People's Republic of China.

Department of Emergency, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China.

出版信息

Dig Dis Sci. 2017 Mar;62(3):639-651. doi: 10.1007/s10620-016-4424-7. Epub 2016 Dec 29.

DOI:10.1007/s10620-016-4424-7

PMID:28035546

Abstract

PURPOSE

The CD8CD28/CD8CD28 T lymphocyte balance is vital for human ulcerative colitis (UC) but has not been defined in experimental colitis. This investigation will try to identify the changes that occur in the CD8CD28/CD8CD28 T lymphocyte balance during the progression of trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats.

METHODS

The frequencies of blood CD8CD28 and CD8CD28 T lymphocytes were detected in the rats belonging to the normal, model, and treated groups on five days using flow cytometry. The treated rats were administered with mesalazine and were euthanized after a 14-day treatment, as were the normal and model rats. The sensitivity and specificity of the CD8CD28/CD8CD28 T lymphocyte balance in diagnosing early colitis were analyzed by receiver operating characteristics (ROC) curves. The frequencies of CD8CD28 and CD8CD28 T lymphocytes in the colon tissue were tested via immunofluorescence. ELISA was used to measure the levels of the cytokines. Immunohistochemistry and Western blotting were used to detect the colonic expression of JAK3, STAT6, NFATc2, and GATA3.

RESULTS

We found that the ratio of CD8CD28/CD8CD28 T lymphocytes decreased, as did the level of interleukin-7, but not IL-12p40, IL-13, or IL-15, in the blood; however, the ratio increased along with JAK3, STAT6, NFATc2, and GATA3 in the colon of the rats with colitis. The changes were effectively reversed through the administration of mesalazine for 13 days. Surprisingly, the balance in the blood could sensitively distinguish rats with early colitis from normal rats.

CONCLUSION

These data show that increase in CD8CD28 T cells in blood and decrease in CD8CD28 T cells in colon are associated with experimental colitis.

摘要

目的

CD8CD28⁺/CD8CD28⁻ T淋巴细胞平衡对人类溃疡性结肠炎（UC）至关重要，但在实验性结肠炎中尚未明确。本研究旨在确定三硝基苯磺酸（TNBS）诱导的大鼠结肠炎进展过程中CD8CD28⁺/CD8CD28⁻ T淋巴细胞平衡的变化。

方法

使用流式细胞术在五天内检测正常组、模型组和治疗组大鼠血液中CD8CD28⁺和CD8CD28⁻ T淋巴细胞的频率。治疗组大鼠给予美沙拉嗪，治疗14天后安乐死，正常组和模型组大鼠也同样处理。通过受试者工作特征（ROC）曲线分析CD8CD28⁺/CD8CD28⁻ T淋巴细胞平衡在诊断早期结肠炎中的敏感性和特异性。通过免疫荧光检测结肠组织中CD8CD28⁺和CD8CD28⁻ T淋巴细胞的频率。采用ELISA法检测细胞因子水平。免疫组织化学和蛋白质印迹法检测结肠中JAK3、STAT6、NFATc2和GATA3的表达。

结果

我们发现，血液中CD8CD28⁺/CD8CD28⁻ T淋巴细胞的比例降低，白细胞介素-7水平也降低，但白细胞介素-12p40、白细胞介素-13或白细胞介素-15水平未降低；然而，在结肠炎大鼠的结肠中，该比例随JAK3、STAT6、NFATc2和GATA3的增加而升高。给予美沙拉嗪13天可有效逆转这些变化。令人惊讶的是，血液中的平衡能够敏感地将早期结肠炎大鼠与正常大鼠区分开来。