Chia-Hui Lin

Global DNA methylation, DNMT1, and MBD2 in patients with rheumatoid arthritis.

OBJECTIVES To investigate the associations of DNA methylation levels and mRNA expressions of DNA cytosine-5-methyltransferase 1 (DNMT1) and methyl CpG-binding domain 2 (MBD2) with rheumatoid arthritis (RA). METHODS The global methylation status of DNA was measured in 65 patients with RA and 64 healthy controls by the ELISA method. DNMT1 and MBD2 mRNA were also detected in 177 RA patients and 95 controls using the quantitative real-time polymerase chain reaction method. RESULTS The global methylation of DNA was significantly decreased in the RA patients compared to the controls (p=0.005, 95% CI=0.0835-0.4503). The patients with RA had higher expressions of DNMT1 and MBD2 mRNA than the controls (p<0.001, 95% CI=-0.0024 to -0.0053 and p<0.001, 95% CI=-0.0079 to -0.0167, respectively). We also found that the MBD2 mRNA level was not related to the disease activity of RA. However, the expression of DNMT1 mRNA tended to be associated with the disease activity of RA (p=0.08). The levels of DNA methylation and DNMT1 mRNA were significantly decreased in the patients with anti-CCP antibody compared with those without (p=0.005, 95% CI=-0.7333 to -0.1373 and p=0.003, 95% CI=-0.0071 to -0.0022, respectively). The differences in the methylation level and expressions of DNMT1 and MBD2 were not significant between the patients treated with and without anti-TNFα biological agents (Enbrel or Humira). CONCLUSION This study demonstrated that the RA patients have a significantly lower level of DNA methylation than the controls. Moreover, RA patients have higher expressions of DNMT1 and MBD2 mRNA. The anti-TNFα biological agents do not seem to affect DNA methylation and mRNA expressions of DNMT1 and MBD2 in RA patients.

Killer cell immunoglobulin-like receptor gene's repertoire in rheumatoid arthritis

Objective: To investigate the role of the killer cell immunoglobulin‐like receptor (KIR) genes repertoire in the pathogenesis of rheumatoid arthritis (RA) in Taiwan. Methods: KIR genotypes were determined in 122 patients with RA and 96 healthy controls by the sequence‐specific primer polymerase chain reaction (SSP‐PCR) method. Human leucocyte antigen (HLA)‐C genotyping was also performed simultaneously in 72 patients and 66 controls by the SSP‐PCR method. Results: The total carriage frequency of KIR 2DS4 regardless of corresponding HLA‐Cw4 was significantly increased in RA patients compared with controls [p<0.001, odds ratio (OR) = 1.9, 95% confidence interval (CI) = 1.1–3.4, Pc<0.01]. The total carriage frequency of KIR 2DL1 regardless of corresponding HLA‐C also tended to be increased in RA patients (p<0.02, OR = 2.1, 95% CI = 1.2–3.9, Pc = not significant). The frequency of KIR 2DS4 with corresponding HLA‐Cw4 was increased in RA patients in comparison with controls (p = 0.02, OR = 3.2, 95% CI = 1.1–9.4). Moreover, the association of RA with KIR 2DS4 depended on the presence of the corresponding HLA‐Cw4. Conclusions: KIR 2DS4 may be a risk factor for susceptibility to RA in Taiwan.

IkBα Promoter Polymorphisms in Patients with Primary Sjögren’s Syndrome

IntroductionTo investigate the association of IkBα promoter polymorphisms with the development of primary Sjögren’s syndrome in Taiwan, 98 patients with primary Sjögren’s syndrome and 110 unrelated healthy controls were enrolled in this study.Materials and MethodsThe IκBα −881 A/G, IκBα −826 C/T, IκBα −550 A/T, IκBα −519 C/T, and IκBα −297 C/T polymorphisms were determined by the methods of polymerase chain reaction/restriction fragment length polymorphism.ResultsThis study demonstrated that the genotype frequencies of IκBα −826 C/T and IκBα −826 T/T, in comparison with that of IκBα −826 C/C, were significantly higher in the patients with primary Sjögren’s syndrome than in the controls. The allele frequency of IκBα −881 G was significantly decreased in the patients with primary Sjögren’s syndrome compared with that of the controls. In contrast, the allele frequency of IκBα −826 T was significantly higher in the patients with primary Sjögren’s syndrome than in the controls. The similar findings could also be found in the allele carriage frequencies. The patients with primary Sjögren’s syndrome had lower allele carriage frequencies of IκBα −881 G and IκBα −826 C, and a higher allele carriage frequency of IκBα −826 T. We also found that the estimated haplotype frequency of IκBα −881A-826T-550A-519C-297C was significantly increased in the patients with primary Sjögren’s syndrome in comparison with that of the controls.DiscussionThis study demonstrated that the IkBα −826T allele and IkBα −881A-826T-550A-519C-297C haplotype were associated with susceptibility to primary Sjögren’s syndrome in Taiwan. However, these findings may not be disease-specific but may be related to inflammatory responses.

IκBα Promoter Polymorphisms in Patients with Systemic Lupus Erythematosus

To investigate the associations of IκBα gene polymorphisms with the development and clinical manifestations of systemic lupus erythematosus (SLE), 110 patients with SLE and 120 unrelated healthy controls were enrolled in this study. The IκBα −881 A/G, −826 C/T, −550 A/T, −519 C/T, and −297 C/T polymorphisms were determined by the polymerase chain reaction/reaction fragment length polymorphism method. The genotype frequency of IκBα −826 C/T in the patients with SLE was significantly higher than that of the controls (p = 0.003, OR = 2.2, 95% CI = 1.3–3.9). The SLE patients also have significantly higher carriage rate of IκBα −826 T than the controls (p = 0.01, OR = 2.0, 95% CI = 1.2–3.4). We also found that the estimated haplotype frequency of IκBα −881A −826T −550A −519C −297C was significantly increased in the patients with SLE in comparison with that of the controls. This study also demonstrated that the association of IκBα −826 T with SLE was independent of HLA-DR15, which is associated with susceptibility to SLE in Taiwan. Moreover, a synergistic effect could also be found between IκBα −826 T and HLA-DR15. IκBα −826 T is associated with the development of SLE in Taiwan. The IκBα −881A −826T −550A −519C −297C haplotype is also associated with susceptibility to SLE. This study also demonstrated that IκBα -881G was associated with the occurrence of vasculitis in SLE patients. IκBα −550T might be a protective factor for the development of malar rash.

Polymorphisms of Genes for Programmed Cell Death 1 Ligands in Patients with Rheumatoid Arthritis

To investigate the role of ligands for programmed cell death 1 (PD-L) in the pathogenesis of rheumatoid arthritis (RA), 129 patients with RA and 125 unrelated healthy controls were enrolled in this study. The PD-L1 and PD-L2 polymorphisms were determined by the method of polymerase chain reaction (PCR)/direct sequencing or PCR/reaction fragment length polymorphisms. The genotype distributions of PD-L1 6777 C/G were not significantly different between the patients with RA and healthy controls. There was also no significant difference in the allele frequencies of PD-L1 6777 C/G polymorphisms between the patients with RA and controls. Similar findings could also be found in the phenotypes and alleles frequencies of PD-L2 47103 C/T and 47139 T/C polymorphisms between the patients with RA and controls. The patients with PD-L1 6777 G had higher prevalence of rheumatoid nodule in comparison with those without PD-L1 6777 G (p = 0.005, OR = 4.0, 95% CI = 1.5–10.9). In contrast, the PD-L2 47103 C/T and 47139 T/C polymorphisms were not related to the occurrence of rheumatoid nodule. This study demonstrated that the PD-L1 and PD-L2 polymorphisms were not associated with susceptibility to RA in Taiwan. PD-L1 6777 G was associated with the prevalence of rheumatoid nodule.

IκBα promoter polymorphisms in patients with Behcet's disease

To investigate the role of IκBα promoter polymorphisms in the development of Behçet’s disease, eighty-six patients with Behçets disease and 120 healthy controls were enrolled in this study. The IκBα; -881A/G, -826C/T, -550A/T, -519C/T, and -297C/T polymorphisms were measured by the method of polymerase chain reaction/ restriction fragment length polymorphism. This study demonstrated that the genotype frequencies of IκBα -826C/T and -826T/T were significantly higher in the patients with Behçets disease than in the controls. Both in the dominant and in the recessive models, the patients with Behçets disease have higher frequencies of the IκBα -826T containing genotype than the controls. The allele frequency of IκBα -826T was significantly increased in the patients with Behçet’s disease. The frequencies of the IκBα -881A -826T -550A -519C -297C and IκBα -881A -826T -550A -519T -297C haplotypes were significantly higher in the patients with Behçet–s disease than in the controls. In contrast, the haplotype frequency of IκBα -881A -826C -550A -519C -297C in the patients with Behçet’s disease was significantly decreased. This study also revealed that the Behçet’s disease patients with IκBα -826T/T have higher prevalence of skin lesions than those without IκBα -826T/T. In summary, the IκBα -826T allele, IκBα -881A -826T -550A -519C -297C and IκBα -881A -826T -550A -519T -297C haplotypes might be associated with susceptibility to Behçet’s disease. The IκBα -826T/T genotype was related to the development of skin lesions in the patients with Behçets disease.

Inhibitor IκBα Promoter Functional Polymorphisms in Patients with Rheumatoid Arthritis

IntroductionRheumatoid arthritis (RA) is a chronic inflammation disease that may involve extra-articular organs in addition to joints. Many proinflammatory cytokines are involved in the inflammatory process of RA. IκBα conjugates with NF-κB and is a key player in regulation of the inflammatory process. We carried out experiments to define the effect of different promoter polymorphisms on the transcriptional activities of IκBα promoter and the development of RA.MethodsDifferent IκBα promoter reporters were constructed and were examined in human mononuclear cells, THP-1 cells. One hundred forty patients and 115 healthy controls were recruited from the Kaohsiung Medical University Hospital.ResultsThe activities of IκBα promoter constructs with -826C, -550A, -519T, and -826T, -550A, -519T genotypes were expressed at one half the activity level of other constructs. Promoter constructs containing the sites -550A/T and -519T had a reduced risk of rheumatoid arthritis. The odds ratio of -826C/T genotype was significantly associated with an increase of risk in causing rheumatoid arthritis, whereas -826T/T genotype was associated only with a slightly increased risk of RA, but without statistical significance (odds ratio = 1.2; 95% confidence interval, 0.4–3.8).ConclusionThe increase of T allele was associated with a significant increased risk and the tendency to the pathogenesis of RA. The association between IκBα promoter polymorphisms and disease severity of rheumatoid arthritis is partly due to different transcriptional activities of IκBα promoter and the activation of NF-κB.

IκBαPromoter Polymorphisms in Patients with Behçet’s Disease

To investigate the role of IκBα promoter polymorphisms in the development of Behçet’s disease, eighty-six patients with Behçet’s disease and 120 healthy controls were enrolled in this study. The IκBα -881A/G, -826C/T, -550A/T, -519C/T, and -297C/T polymorphisms were measured by the method of polymerase chain reaction/ restriction fragment length polymorphism. This study demonstrated that the genotype frequencies of IκBα -826C/T and -826T/T were significantly higher in the patients with Behçet’s disease than in the controls. Both in the dominant and in the recessive models, the patients with Behçet’s disease have higher frequencies of the IκBα -826T containing genotype than the controls. The allele frequency of IκBα -826T was significantly increased in the patients with Behçet’s disease. The frequencies of the IκBα -881A -826T -550A -519C -297C and IκBα -881A -826T -550A -519T -297C haplotypes were significantly higher in the patients with Behçet’s disease than in the controls. In contrast, the haplotype frequency of IκBα -881A -826C -550A -519C -297C in the patients with Behçet’s disease was significantly decreased. This study also revealed that the Behçet’s disease patients with IκBα -826T/T have higher prevalence of skin lesions than those without IκBα -826T/T. In summary, the IκBα -826T allele, IκBα -881A -826T -550A -519C -297C and IκBα -881A -826T -550A -519T -297C haplotypes might be associated with susceptibility to Behçet’s disease. The IκBα -826T/T genotype was related to the development of skin lesions in the patients with Behçet’s disease.

IkBα promoter polymorphisms in patients with ankylosing spondylitis

The purpose of this study is to investigate the association of IκBα with the development of ankylosing spondylitis (AS) in Taiwan. One hundred and fifty-four patients with AS and 112 unrelated healthy controls were enrolled in this study. The IκBα-881A/G, -826C/T, -550A/T, -519C/T, and -297C/T polymorphisms were determined by the polymerase chain reaction/restriction fragment length polymorphism method. This study demonstrated that the genotype frequencies of IκBα-826C/T and -826T/T, and allele frequencies of IκBα-826T were significantly higher in the patients with AS than in the controls. We also found that the estimated haplotype frequencies of IκBα-881A -826T -550A -519C -297C and IκBα-881A -826C -550A -519T -297C were significantly increased in the patient with AS in comparison with that of the controls. In contrast, the estimated haplotype frequency of IκBα-881A -826C -550A -519C -297C was significantly decreased in the patients with AS. This study demonstrates that IκBα-826T is associated with the development of AS. Furthermore, the IκBα-881A -826T -550A -519C -297C and IκBα-881A -826C -550A -519T -297C haplotypes are related to susceptibility to AS in Taiwan.

Solute Carrier Family 11 Member A1 Gene Polymorphisms in Reactive Arthritis

To investigate the role of SLC 11A1 polymorphisms in the development of reactive arthritis, 91 patients with reactive arthritis and 163 healthy controls were enrolled in this study. The SLC 11A1 polymorphisms were determined by the method of polymerase chain reaction/restriction fragment length polymorphism. The genotype distributions of SLC 11A1 274, 823, 1703, and 1729+ 55 del 4 were significantly different between the patients with reactive arthritis and controls. The genotype frequency of SLC 11A1 274C/C was significantly decreased in the patients with reactive arthritis when compared with that of the controls. In contrast, the SLC 11A1 274C/T showed a significant association with reactive arthritis. The patients with reactive arthritis have a significantly higher frequency of SLC 11A1 823C/C than the controls. However, SLC 11A1 823T/T was resistant to the development of reactive arthritis. The allele frequencies of SLC 11A1 274T and 823C were significantly increased in the patients with reactive arthritis in comparison with those of the controls, independent of HLA-B27. On the contrary, the allele frequencies of SLC 11A1 274C and 823T were significantly decreased in the patients with reactive arthritis. The estimated haplotype frequency of SLC 11A1 274C 823T 1703G 1729+55del 4 TGTG+ was significantly decreased in the patients with reactive arthritis when compared with that of the controls. In contrast, the estimated haplotype frequency of SLC 11A1 274T 823C 1703G 1729+55 del 4 TGTG+ was significantly increased in the patients with reactive arthritis. This study shows that the SLC 11A1 274T and 823C alleles are associated with susceptibility to reactive arthritis independently of HLA-B27 in Taiwan. The SLC 11A1 274T 823C 1703G 1729+55 del 4 TGTG+ haplotype is associated with the development of reactive arthritis in Taiwan. In contrast, the SLC 11A1 274C 823T 1703G 1729+55 del 4 TGTG+ haplotype may be a protective factor.