[Association between the synovial expression of cyclic citrullinated peptide and susceptibility variants of HLA-DRB1 shared epitope alleles and PADI 4 gene single nucleotide polymorphisms in patients with rheumatoid arthritis].
[Association between the synovial expression of cyclic citrullinated peptide and susceptibility variants of HLA-DRB1 shared epitope alleles and PADI 4 gene single nucleotide polymorphisms in patients with rheumatoid arthritis].
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DOI：
10.3760/cma.j.issn.0376-2491.2012.23.007
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发表时间：
2012-06
影响因子：
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通讯作者：
Hong-bin Li;Ning Tie;Yong-feng Jia;Lin Shi;Yan Su;Guizhi Zhang;Yong Wang;L. Bai;J. Zhao;Jing Wang;Zhen Xiao
中科院分区：
文献类型：
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作者：
Hong-bin Li;Ning Tie;Yong-feng Jia;Lin Shi;Yan Su;Guizhi Zhang;Yong Wang;L. Bai;J. Zhao;Jing Wang;Zhen Xiao
目的分析类风湿关节炎（RA）患者滑膜环瓜氨酸肽（CCP）表达与HLA-DRB 1共享表位（SE）等位基因和/或肽基精氨酸脱亚胺酶4（PADI 4）基因单核苷酸多态性（SNP）易感性的关系。方法2008年10月至2011年12月，入组53例RA患者和42例对照。用pHEN 2噬菌粒重组抗体展示系统制备抗CCP单链抗体（6×His taged single chain fragment V，ScFv），免疫组化法检测RA滑膜组织中CCP的表达。采用逆转录cDNA测序法对PADI 4 SNP进行基因分型，TA克隆测序法定位杂合子DNA单倍型。采用序列特异性引物-聚合酶链反应（SSP-PCR）分析HLA-DRB 1 SE等位基因。结果RA组滑膜CCP阳性表达率为76.9%，对照组为11.4%，两组比较差异有显著性（P < 0.01）。PADI 4_89 G+和PADI 4_104 T+两个单核苷酸多态性位点的频率在各组间差异有统计学意义（P < 0.05）。与主要单倍型相比，只有这两个次要等位基因与RA易感性增加相关（OR = 3.67和2.53，P < 0.05）。SE+等位基因与RA易感性密切相关（OR = 5.57，P < 0.01）。RA滑膜中CCP的表达与SE+等位基因、仅2个次要SNP（PADI 4_89 G+和PADI 4_104 T+）以及组合强烈相关。血清抗CCP滴度与SE+等位基因、PADI 4_104T+、SE+/PADI 4_89 G+和SE+/PADI 4_104T+单倍型显著相关。结论RA滑膜组织CCP表达及抗CCP抗体的产生与SE等位基因和/或PADI 4基因的某些SNP密切相关。
OBJECTIVE To analyze the association between the synovial expression of cyclic citrullinated peptide (CCP) and susceptibility variants of HLA-DRB1 shared epitope (SE) alleles and/or peptidylarginine deiminase 4 (PADI4) gene single nucleotide polymorphisms (SNP) in patients with rheumatoid arthritis (RA). METHODS From October 2008 to December 2011, 53 RA patients and 42 controls were enrolled. The expression of CCP in RA synovial tissues was detected by immunohistochemical assay with 6×His tagged anti-CCP single chain fragment V (ScFv) antibodies generated by pHEN2 phagemid recombinant antibodies display system. PADI4 SNP was genotyped by reverse transcription cDNA sequencing and heterozygote was DNA haplotype was mapped by TA clone sequencing. HLA-DRB1 SE alleles were analyzed by sequence specific primer-polymerase chain reaction (SSP-PCR). RESULTS The prevalence of synovial CCP expression was significantly different between RA group and the control (76.9% and 11.4% respectively, P < 0.01). The frequencies of 2 SNPs (PADI4_89 G+ and PADI4_104 T+) varied significantly between the groups(P < 0.05). Compared with the major haplotypes, only these two minor alleles were associated with the increased RA susceptibility (OR = 3.67 and 2.53, P < 0.05). SE+ alleles was strongly associated with RA susceptibility (OR = 5.57, P < 0.01). The synovial expression of CCP in RA was strongly associated with SE+ alleles, only 2 minor SNPs (PADI4_89 G+ and PADI4_104T+) and the combination. Serum anti-CCP titers were significantly associated with SE+ alleles, PADI4_104T+, SE+/PADI4_89 G+ and SE+/PADI4_104T+ haplotype. CONCLUSION The synovial expression of CCP and the generation of anti-CCP antibodies are strongly associated with SE alleles and/or certain PADI4 gene SNP in RA.