Matsuzaki Koji, Hokari Ryota, Kato Shingo, Tsuzuki Yoshikazu, Tanaka Hirofumi, Kurihara Chie, Iwai Atsuhiro, Kawaguchi Atsushi, Nagao Shigeaki, Itoh Kazuro, Nagata Kinya, Miura Soichiro
Second Department of Internal Medicine, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan.
J Gastroenterol Hepatol. 2003 Sep;18(9):1081-8. doi: 10.1046/j.1440-1746.2003.03088.x.
The pathogenesis of ulcerative colitis (UC) is unclear, but abnormal infiltration of T lymphocytes in the colonic mucosa has been implicated in the mucosal tissue damage. The abnormal cytokine production because of a T helper (h)1/Th2 imbalance may play an important role in continuing inflammation in the colonic mucosa. In the present study, the expression of chemokine receptor 5 (CCR5) as a Th1 marker and a chemoattractant receptor-homologs molecule expressed on Th2 cells (CRTH2) were investigated in order to analyze impaired Th1/Th2 responses in the colonic mucosa of UC patients.
Tissue samples were obtained by colonic biopsies from patients with UC or colonic polyps, with informed consent. Immunohistochemical analysis was performed on periodate, lysine-paraformaldehyde-fixed serial cryostat sections using the labeled streptavidin biotin method. Monoclonal antibodies against CD4, CCR5 or CRTH2 were used as primary antibodies. The number of cells expressing CD4, CCR5 or CRTH2 per unit area was calculated by using an image analyzer.
In the patients with UC, the numbers of CD4- and CCR5-positive cells were significantly increased in inflamed mucosa, and appeared to be correlated with the disease activity. The infiltration of CRTH2-positive cells was predominantly observed in the mildly inflamed or the margin of inflamed mucosa of UC patients.
There is a possibility that Th1 responses significantly occur in colonic mucosa with severe inflammation, while Th2 responses mainly occur with mild inflammation in UC patients. The Th1/Th2 imbalance in colonic mucosa may be related to the disease progression of UC.
溃疡性结肠炎（UC）的发病机制尚不清楚，但结肠黏膜中T淋巴细胞的异常浸润与黏膜组织损伤有关。由于辅助性T（h）1/Th2失衡导致的细胞因子异常产生可能在结肠黏膜持续炎症中起重要作用。在本研究中，为了分析UC患者结肠黏膜中Th1/Th2反应受损情况，对作为Th1标志物的趋化因子受体5（CCR5）以及Th2细胞上表达的趋化因子受体同源分子（CRTH2）的表达进行了研究。
在获得知情同意后，通过结肠活检从UC患者或结肠息肉患者获取组织样本。使用标记链霉亲和素生物素法对经高碘酸盐、赖氨酸-多聚甲醛固定的连续低温恒温器切片进行免疫组织化学分析。抗CD4、CCR5或CRTH2的单克隆抗体用作一抗。使用图像分析仪计算每单位面积表达CD4、CCR5或CRTH2的细胞数量。
在UC患者中，炎症黏膜中CD4和CCR5阳性细胞数量显著增加，且似乎与疾病活动度相关。CRTH2阳性细胞的浸润主要见于UC患者轻度炎症或炎症黏膜边缘。
在UC患者中有可能在严重炎症的结肠黏膜中显著发生Th1反应，而在轻度炎症时主要发生Th2反应。结肠黏膜中的Th1/Th2失衡可能与UC的疾病进展有关。
