Department of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei Province, China.
J Dig Dis. 2010 Dec;11(6):369-75. doi: 10.1111/j.1751-2980.2010.00462.x.
Our aim was to investigate the expression of cytotoxic T lymphocyte-associated molecule 4 (CTLA-4) in ulcerative colitis (UC) and to evaluate the effect of CTLA-4 gene -1661A/G polymorphism on CTLA-4 expression and transcription.
A total of 20 UC patients and 22 healthy controls matched by age and sex were enrolled at Zhongnan Hospital of Wuhan University in central China. The CTLA-4 -1661A/G polymorphism was genotyped by the polymerase chain reaction-restriction fragment length polymorphism method. A Western blot analysis was performed to determine the full length CTLA-4 (flCTLA-4) protein expression in the peripheral blood of the UC patients. Serum-soluble CTLA-4 (sCTLA-4) levels were measured by enzyme-linked immunosorbent assay. CTLA-4-1661G mutant promoter transcription function was analyzed by site-directed PCR-based mutagenesis.
CTLA-4 protein expression on CD4(+) T cells in UC patients was lower than that in the healthy controls (P < 0.001) while serum sCTLA-4 in the UC patients was significantly higher than that in the healthy controls (P < 0.001). No correlation was found between flCTLA-4 and sCTLA-4 expression levels and the -1661 A/G polymorphism of the CTLA-4 gene. Meanwhile, CTLA-4 -1661 allele A had no significant impact on the promoter activity compared with allele G (P > 0.05).
CTLA-4 expressions were aberrant in UC patients compared with the healthy controls. CTLA-4 -1661A/G polymorphism had no significant impact on CTLA-4 expression and transcription in the peripheral CD4 T cells of UC patients.
本研究旨在探讨细胞毒性 T 淋巴细胞相关抗原 4(CTLA-4)在溃疡性结肠炎(UC)中的表达,并评估 CTLA-4 基因-1661A/G 多态性对 CTLA-4 表达和转录的影响。
本研究纳入了 20 例 UC 患者和 22 名年龄和性别相匹配的健康对照者,均来自中国中部的武汉大学中南医院。采用聚合酶链反应-限制性片段长度多态性方法检测 CTLA-4-1661A/G 多态性。采用 Western blot 分析检测 UC 患者外周血中全长 CTLA-4(flCTLA-4)蛋白的表达。采用酶联免疫吸附试验检测血清可溶性 CTLA-4(sCTLA-4)水平。采用基于定点 PCR 诱变的方法分析 CTLA-4-1661G 突变启动子的转录功能。
UC 患者 CD4+T 细胞上 CTLA-4 蛋白的表达低于健康对照组(P<0.001),而 UC 患者血清 sCTLA-4 水平明显高于健康对照组(P<0.001)。flCTLA-4 与 sCTLA-4 表达水平与 CTLA-4 基因-1661A/G 多态性之间无相关性。同时,与 CTLA-4-1661 等位基因 G 相比,等位基因 A 对启动子活性没有显著影响(P>0.05)。
与健康对照者相比,UC 患者 CTLA-4 表达异常。CTLA-4-1661A/G 多态性对 UC 患者外周血 CD4+T 细胞中 CTLA-4 的表达和转录没有显著影响。