细胞毒T淋巴细胞相关抗原4基因多态性与溃疡性结肠炎

目的炎症性肠病的发病与T细胞过度活化有关,细胞毒T淋巴细胞相关抗原4（CTLA-4）是重要的T细胞活化负性调节因子.本课题研究CTLA-4基因启动子区-1722位点（T/C）及-1661位点（A/G）多态性与中国汉族人群中溃疡性结肠炎（UC）的相关性.方法采用PCR-限制性片段长度多态性方法,对87例中国汉族UC患者和116例正常对照者进行CTLA-4基因-1722位点和-1661位点多态性检测.结果UC患者CTLA-4基因-1661位点A/G＋G/G基因型频率,-1661位点G等位基因频率显著高于正常对照组（34.5%比15.5%,P=0.002,OR=2.865,95%CI=1.467～5.596;19.0%比8.2%,P=0.002,OR=2.624,95%CI=1.435～4.796）;而在-1722位点的基因型频率、等位基因频率与对照组比较差异无统计学意义（P＞0.05）.结论CTLA-4基因启动子区-1661位点G等位基因与中国汉族UC存在显著相关性.     

A functional polymorphism of the NFKB1 promoter is not associated with ulcerative colitis in a Spanish population

BACKGROUND: This study investigated the influence of the NFKB1-94ins/delATTG in the susceptibility/phenotype to ulcerative colitis. METHODS: We analyzed the distribution of -94ins/delATTG NFKB1 in 258 patients and 264 healthy controls from southern Spain by a polymerase chain reaction-fluorescent method. RESULTS: The genotype and allele frequencies of -94ins/delATTG did not significantly differ between patients and controls. In fact, the frequency of the -94delATTG allele was almost identical in both groups (34.8% and 35.4%, respectively), and the del/del genotype was underrepresented in UC patients (11.2% versus 14%). In addition, no association of this polymorphism was found with any of the clinical parameters analyzed. CONCLUSION: These results suggest that the NFKB1 -94ins/delATTG gene variation, previously associated with UC susceptibility in North Americans, does not influence either susceptibility or phenotype of UC in the Spanish population.     

MICB0106 gene polymorphism is associated with ulcerative colitis in central China

Background  

The highly polymorphic nonclassical MHC class I chain-related genes A and B (MICA and MICB) encode stress-inducible glycoproteins expressed on various epithelial cells including intestinal epithelial cells. MICA and MICB gene polymorphisms and expressions are associated with autoimmune diseases but not known in ulcerative colitis (UC).     

Promoter polymorphism of cyclooxygenase-1 (COX1) is not associated with ulcerative colitis in the Japanese population

BACKGROUND/AIMS: Non-steroidal anti-inflammatory drugs (NSAIDs) aggravate or ameliorate clinical disease activity in patients with ulcerative colitis (UC). UC is associated with increased local production of prostanoids. This study attempted to clarify the relationship between cyclooxygenase-1 (COX1) gene polymorphisms (T-1676C and A-842G/C50T) and ulcerative colitis in a Japanese population. METHODOLOGY: The study was performed on 108 patients with UC (mean age: 39.1 years, M:F=56:52) and 293 healthy controls (mean age: 49.0 years, M:F=161:132). The PCR-SSCP method was employed to detect COX1 polymorphisms using DNA extracted from peripheral blood cells. RESULTS: No A-842G/C50T polymorphism was detected in all 401 Japanese subjects. The frequency of -1676C allele of COX1 gene in HC group and UC group was 48.8% and 48.1%, respectively. T-1676C genotypes of COX1 did not show significant association with UC risk. In addition, these genotypes were not also associated with the clinicopathological parameters of UC. CONCLUSIONS: This research suggests that COX1 promoter polymorphisms (T-1676C and A-842G/ C50T) may not be associated with the risk of developing ulcerative colitis in a Japanese population.     

Ulcerative colitis is associated with a promoter polymorphism of lipopolysaccharide receptor gene,CD14

BACKGROUND: Inflammatory bowel disease (IBD) is a multifactorial disease with a significant genetic background. Evidence is accumulating that molecules such as CD14, which interact with luminal bacterial constituents, are involved in the pathogenesis. It has recently been shown that the T allele of the 5'-flanking region of the CD14 gene at position -159 is related to high expression of CD14. In further exploring the genetic background of IBD, we investigated this novel polymorphism of CD14 gene in patients with ulcerative colitis or Crohn disease. METHODS: DNA was obtained from 101 patients with ulcerative colitis, 82 with Crohn disease and 123 healthy controls. All were typed for the promoter polymorphism of the CD14 gene at position -159 by restriction fragment length polymorphism analysis. Serum samples were obtained from 105 healthy controls and serum sCD14 levels were measured. RESULTS: T allele frequencies were 57.4%, 48.2% and 44.7% in ulcerative colitis, Crohn disease and healthy controls, respectively. The T allele and T/T genotype frequencies were significantly higher in ulcerative colitis patients than in healthy controls (P = 0.0074, OR = 1.67, 95% CI = 1.15-2.42, P = 0.022, OR= 1.96 95% CI: 1.10-3.48, respectively). The sCD14 level was significantly higher in TT genotype populations than CC (P = 0.0205). CONCLUSIONS: The promoter polymorphism of the CD14 gene at -159T plays a significant role in regulating the CD14 expression and is positively associated with ulcerative colitis, and this polymorphism may confer a genetic predisposition to ulcerative colitis. The results also support the concept that bacterial constituents may be involved in the pathogenesis of ulcerative colitis.     

Susceptibility to severe ulcerative colitis is associated with polymorphism in the central MHC gene IKBL

BACKGROUND & AIMS: IKBL gene lies telomeric of the tumor necrosis factor cluster in the central major histocompatibility complex and carries a structural polymorphism at position +738. In the Spanish white population, we found the IKBL+738(C) allele in haplotypes carrying either HLA-DRB1(*)1501 or -DRB1(*)0103. Because these HLA class II alleles may confer susceptibility to ulcerative colitis, we investigated an association between IKBL+738(C) and this disease. METHODS: DNA-based techniques were used to type individual alleles of HLA-DRB1 and IKBL+738. The frequencies of these alleles were compared among ethnically matched populations comprising 155 patients and 298 controls. RESULTS: IKBL+738(C) allele was exclusively increased in patients with extensive and/or intractable disease. HLA-DRB1(*)0103 was the only HLA-DRB1 allele to be significantly increased in frequency in patients with UC compared with controls. It was found in patients with extensive and distal disease. In the HLA-DRB1(*)0103-negative population, patients with extensive disease still had a significant association with IKBL+738(C). The difference between the 2 groups of patients was statistically significant (13.7% vs. 1.7% in patients with distal disease; odds ratio, 9.25; P = 0.01). CONCLUSIONS: HLA-DRB1(*)0103 is associated with susceptibility to ulcerative colitis, and IKBL+738(C) marks a propensity to extensive and more severe disease.     

Nrf2 gene promoter polymorphism and gastric carcinogenesis

BACKGROUND/AIMS: Three gene polymorphisms of Nrf2, which regulate the expression of detoxifying and antioxidant genes, have been identified. We attempted to clarify the relationship of these polymorphisms with the carcinogenesis in the stomach. METHODOLOGY: The study was performed in 209 patients with gastric cancer and 198 patients with no evidence of gastric malignancies on upper gastroduodenal endoscopy. We employed PCR-SSCP method to detect gene polymorphisms. RESULTS: Overall, both polymorphisms at position of -686/-684 and -650 were not significant risk factors of carcinogenesis in the stomach. However, the -686/-684 A/G allele carrier had a significantly reduced risk for gastric carcinogenesis (p = 0.022), especially of diffuse type (p = 0.020), in H. pylori-negative cases. The activity and inflammation scores in Nrf2 -686/-684 A/G carriers were significantly lower than those in the non-A/G carriers (p = 0.038 and p = 0.019, respectively). CONCLUSIONS: The -686/-684 haplotype of Nrf2 gene may be associated with the development of gastric inflammation and with gastric carcinogenesis without the influence of H. pylori infection, although overall association with gastric carcinogenesis seems to be none.     

Susceptibility to refractory ulcerative colitis is associated with polymorphism in the hMLH1 mismatch repair gene

The hMLH1 gene lies in the linkage susceptibility region to inflammatory bowel disease (IBD) on 3p21. A single nucleotide polymorphism, 655A>G, in exon 8 of the gene causes an I219V change in the MLH1 protein. To test whether hMLH1 may confer susceptibility to ulcerative colitis (UC), we investigated an association between the 655A>G polymorphism and the disease. DNA-based technologies were used to analyze the 655A>G polymorphism in 201 UC patients and 126 healthy ethnically matched controls. The comparison of the allelic frequencies of the 655A>G polymorphism in UC patients and healthy controls did not show significant differences. However, genotype frequencies at the hMLH1 655 position were found to be significantly different when patients with and without refractory UC were compared. This was mainly attributable to a higher level of homozygosity for the G allele in refractory UC patients. Almost 5 times as many (4.9 times) refractory UC patients carried the GG genotype compared with nonrefractory patients (P < 0.0001). The present study provides evidence that the hMLH1 gene is involved in genetic susceptibility to refractory UC. If confirmed by other studies, the GG genotype at position 655 of the hMLH1 gene may represent a useful predictive factor for the clinical management of UC patients.     

Severity of ulcerative colitis is associated with a polymorphism at diamine oxidase gene but not at histamine N-methyltransferase gene

AIM:To analyse the role of two common polymorphismsin genes coding for histamine metabolising enzymes asit relates to the risk to develop ulcerative colitis(UC)andthe clinical course of these patients.METHODS:A cohort of 229 unrelated patients withUC recruited from a single centre and 261 healthyvolunteers were analysed for the presence of Thr105Ileand His645Asp amino acid substitutions at histamineN-methyltransferase(HNMT)and diamine oxidase(ABP1)enzymes,respectively,by amplification-restriction proce-dures.All patients were phenotyped and followed up forat least 2 years(mean time 11 years).RESULTS:There were no significant differences in thedistribution of ABP1 alleles between ulcerative colitispatients and healthy individuals[OR(95% CI)for vari-ant alleles=1.22(0.91-1.61)].However,mutated ABP1alleles were present with higher frequency among the58 patients that required immunosuppresive drugs[OR(95 % CI)for carriers of mutated alleles 2.41(1.21-4.83;P=0.006)],with a significant gene-dose effect(P=0.0038).In agreement with the predominant role ofABP1 versus HNMT on local histamine metabolism in hu-man bowel,the frequencies for carriers of HNMT geno-types or mutated alleles were similar among patients, regardless clinical evolution,and control individuals.CONCLUSION:The His645Asp polymorphism of thehistamine metabolising enzyme ABP1 is related to severityof ulcerative colitis.     

Vitamin D receptor gene polymorphism is associated with Graves' disease in the Japanese population

Susceptibility to Graves' disease (GD), which is determined by environmental and genetic factors, is conferred by genes in the human leukocyte antigen (HLA) and genes unlinked to HLA, including the CTLA-4 gene. We recently described the association of GD with the vitamin D receptor (VDR) exon 2 initiation codon (VDR-FOK:I) polymorphism. An association of some VDR genotypes with osteoporosis, primary hyperparathyroidism, and some autoimmune diseases, such as insulin-dependent diabetes mellitus and multiple sclerosis, has been reported. We investigated the distribution of VDR gene polymorphism in 180 Japanese patients with GD (48 males and 132 females) and 195 controls (67 males and 128 females). A VDR allelic polymorphism was assessed by BSM:I endonuclease restriction after specific PCR amplification. Genotypic polymorphism was clearly defined as BB (no restriction site on both alleles), bb (restriction site on both alleles), or Bb (heterozygous). The distribution of genotype frequencies differed between patients with GD and controls (chi(2) = 7.53; 2 degrees of freedom; P: = 0.023). The relative risk conferred by at least 1 B allele (BB or Bb) was 1.5. We also found an association between VDR-APA:I polymorphism and GD. No relation was detected between this polymorphism and the VDR-FOK:I polymorphism in the patients. The present results support the association of the VDR gene with GD in Japanese by showing that the VDR gene could be a non-HLA-linked gene predisposing an individual to GD. The role of the VDR gene polymorphism should be further studied in other populations, and the distribution of other polymorphisms, such as the polyadenylase polymorphism further down the VDR 3'-untranslated region, should be studied in terms of GD susceptibility.     

A NOD2 gene polymorphism is associated with the prevalence and severity of chronic obstructive pulmonary disease in a Japanese population

Background and objective: Genetic background is thought to be one of the risk factors for development of COPD. Recently, it has been proposed that the innate immune system is involved in the pathophysiology of COPD. We hypothesized that polymorphisms in the nucleotide‐binding and oligomerization domain (NOD)1 and NOD2 genes would be associated with the pathogenesis of COPD. In addition, the associations between these single nucleotide polymorphisms (SNPs) and phenotypes of COPD were analysed. Methods: Japanese COPD patients (n = 228) and non‐COPD smokers (n = 101) were recruited from the outpatient clinic at Kyoto University Hospital, Kyoto, Japan. At entry into the study, a blood sample was taken and a pulmonary function test was performed. Genotyping was performed for 6 selected tag SNPs of NOD1 and 5 tag SNPs of NOD2. Further investigations were performed for SNP that were associated with COPD, including baseline gene expression, the relative proportions of splicing variants in whole blood, responses to ligand and enhancement of gene expression in peripheral blood neutrophils stimulated with pro‐inflammatory cytokines. Results: The distribution of NOD2 rs1077861 genotypes differed between Japanese COPD patients and non‐COPD smokers (P = 0.036). This SNP was also associated with a lower FEV₁ % predicted (57.2 ± 1.8 for TT vs 50.8 ± 2.3 for TA/AA, P = 0.03) and DL_CO/V_A (2.89 ± 0.1 in TT vs 2.53 ± 0.14 in TA/AA, P = 0.036) in COPD patients. NOD2 gene expression after stimulation with 10 ng/mL of tumour necrosis factor‐α for 4 h, was increased to a greater extent in TA/AA genotype than in TT genotype peripheral blood neutrophils (P = 0.015). Conclusions: The NOD2 rs1077861 SNP may influence the development and progression of COPD in Japanese subjects.     

Heat shock protein gene 70-2 polymorphism is differentially associated with the clinical phenotypes of ulcerative colitis and Crohn's disease

BACKGROUND AND AIM: A single nucleotide polymorphism in heat shock protein 70-2 (HSP70-2) has been shown to be associated with a severe clinical course in Crohn's disease (CD), but it is not known if such a relationship exists in ulcerative colitis (UC). The aim of the present study was to identify associations between the HSP70-2 polymorphism and the clinical courses of CD and UC in Koreans. METHODS: Restriction fragment length polymorphism analysis was performed for HSP70-2 polymorphisms using the PstI-cleavage site present in the B allele but not in the A allele of the DNA obtained from 101 patients with CD, 144 patients with UC, and 245 age- and sex-matched healthy controls. Study subjects were classified by disease behavior, severity and extent of disease. RESULTS: In CD, multivariate analysis showed that the AA genotype of HSP70-2 polymorphisms was associated with non-perforating disease (OR 10.10, 95% CI 1.66-15.38) and male sex (OR 3.56, 95% CI 1.04-12.23), and that the BB genotype was associated with severe CD (OR 12.03, 95% CI 1.60-101.56). In contrast, multivariate analysis for UC showed that the AA genotype was associated with severe UC (OR 2.02, 95% CI 1.34-3.03). CONCLUSIONS: CD patients with BB genotype of HSP70-2 polymorphism tend to experience a more severe clinical course and allele A is associated with more severe UC. HSP70-2 polymorphism may be used to predict CD and UC phenotypes, which can illuminate immunological differences in CD and UC.     

The endothelial dysfunction in patients with type 2 diabetes mellitus is associated with IL-6 gene promoter polymorphism in Chinese population

The purpose of this study is to examine the effects of IL-6 gene promoter -174G/C and -572G/C polymorphism on endothelial function of Chinese T2DM and normal glucose regulation (NGR) subjects. 512 newly diagnosed T2DM patients and 483 NGR subjects were recruited and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) was performed for the IL-6 gene promoter -174G/C and -572G/C polymorphism. Flow-mediated dilation (FMD) was measured as a non-invasive indicator for endothelial function. The results show that the C allele and CC genotype at -174 of IL-6 gene promoter region was extremely rare in both T2DM and NGR groups; genotypes' and alleles' frequency at -572 of IL-6 gene promoter region is of no difference between T2DM and NGR groups; within T2DM group, higher plasma IL-6 concentration and lower FMD was found in patients with -572 GC (2.36 ± 0.69, 4.23 ± 3.82%) or GG (2.32 ± 0.74, 4.24 ± 3.67%) genotype, compared with patients with CC (2.15 ± 0.62, 5.28 ± 3.94%) genotype. The conclusion of the study is that in comparison with patients of CC genotype, the T2DM patients of -572 GC or GG genotype may have more aggravated endothelial dysfunction (ED) and be at higher risk for coronary artery disease (CAD).     

The alpha 2 type IX collagen gene tryptophan polymorphism is not associated with rheumatoid arthritis in the Japanese population

The aim of this study was to investigate whether the alpha 2 type IX collagen (COL9A2) polymorphism that introduces tryptophan residue into the collagen triple-helix is a marker of susceptibility to, or severity of, rheumatoid arthritis (RA). The study included 749 Japanese patients with RA. One hundred twenty-four unrelated healthy individuals served as the control subjects. The relationship between the COL9A2 gene polymorphism and clinical manifestations of RA was evaluated. For the number of subjects positive for COL9A2 tryptophan polymorphism, there was no statistically significant difference between RA patients and normal controls. Furthermore, we did not detect any association of COL9A2 tryptophan polymorphism with disease status, least erosive subset, more erosive subset, or mutilating disease. The lack of association of COL9A2 tryptophan polymorphism with RA and the clinical findings in our study implies that the polymorphism may not function as a candidate gene marker for screening RA patients.     

Interleukin 6 gene promoter polymorphism is not associated with ankylosing spondylitis

OBJECTIVE: To investigate the possible association between the recently described interleukin 6 (IL-6) promoter polymorphisms at position -174, and susceptibility to ankylosing spondylitis (AS). METHODS: Ninety-two patients with AS, 157 healthy controls, and an additional group of 52 HLA-B27 positive unrelated individuals were included in this study. The -174 polymorphic site in the promoter region of IL-6 gene was typed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: No statistically significant differences were observed when IL-6 promoter genotype and allele distribution between patients with AS and healthy controls were compared. CONCLUSION: Our results suggest the -174 IL-6 polymorphism does not play an important role in susceptibility to AS. Larger studies are needed to provide more conclusive evidence on the role of -174 IL-6 polymorphism in AS.