类风湿关节炎患者滑膜CCP表位表达与HLA—DRB1等位基因、PADI4单核苷酸多态性的相关性分析  

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

作　　者：李鸿斌[1] 铁宁[1] 贾永峰[2] 施琳[2] 苏燕[3] 张桂芝[1] 王勇[1] 白丽杰[1] 赵静[1] 王静[1] 肖镇[1] 

机构地区：[1]内蒙古医学院附属医院风湿免疫科,呼和浩特010050 [2]内蒙古医学院附属医院病理科,呼和浩特010050 [3]内蒙古科技大学包头医学院生物化学与分子生物学教研室

出　　处：《中华医学杂志》2012年第23期1607-1611,共5页National Medical Journal of China

基　　金：基金项目：国家自然科学基金（30760235）;教育部春晖计划（Z2007-1-01008）;内蒙古自然科学基金（200711020909）

摘　　要：目的分析类风湿关节炎（RA）患者滑膜环瓜氨酸肽（CCP）表位表达情况与HIA—DRB1共同表位（SE）等位基因、肽酰基精氨酸脱亚氨酶4（PADI4）基因单核苷酸多态性（SNPs）的相关性。方法对象为2008年10月至2011年12月对内蒙古医学院附属医院风湿免疫科和骨科的53例RA患者和42例对照组患者（外伤者26例，骨关节炎16例）。抗CCP单链抗体（ScFv）通过噬菌体蛋白展示技术构建。以6×His·Tag免疫组化法检测RA关节滑膜的CCP表达情况。PADI4基因SNP通过逆转录合成cDNA测序，杂合子TA载体克隆测序。序列特异性引物PCR（SSP—PCR）法检测HLA—DRB1等位基因型。结果CCP表位在RA组与对照组表达率分别为77．4％和9．5％，差异有统计学意义（P〈0．01）。RA组2个PADI4SNP基因型（PADI4_89G^＋和PADI4_104T^＋）的频率与对照组相比差异有统计学意义（P〈0．05）。携带这2个少见基因型者较常见基因型发生RA的危险性为3．67和2．53倍。携带SE^＋等位基因型者较SE^-者发生RA的危险性为5．57倍。RA滑膜CCP表位表达仅与SE^＋等位基因型和PADI4_89G^＋、PADI4_104T^＋以及两者同时阳性相关。血清CCP滴度与SE^＋、PADI4_104T^＋G^＋，SE^＋／PADI4_89G^＋和SE^＋／PADI4_104T^＋等基因型相关。结论RA患者滑膜CCP表位表达和抗CCP抗体的产生与特定的SE等位基因型和PADI4基因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 ＋ ） variedsignificantly 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.

关 键 词：关节炎 类风湿 HLA抗原 精氨酸酶 

分 类 号：R593.22[医药卫生—内科学]