Ruiz-Noa Yeniley, Hernández-Bello Jorge, Llamas-Covarrubias Mara A, Palafox-Sánchez Claudia A, Oregon-Romero Edith, Sánchez-Hernández Pedro Ernesto, Ramírez-Dueñas Maria Guadalupe, Parra-Rojas Isela, Muñoz-Valle Jose Francisco
Instituto de Investigación en Ciencias Biomédicas, CUCS, Universidad de Guadalajara, Guadalajara, México.
Departamento de Fisiología, Laboratorio de Inmunología, CUCS, Universidad de Guadalajara, México.
J Clin Lab Anal. 2019 Mar;33(3):e22710. doi: 10.1002/jcla.22710. Epub 2018 Nov 6.
CD40 is a costimulatory molecule for B cells, and CD154 is a marker of CD4+ T cells activation. CD40-CD154 interaction promotes pro-inflammatory cytokines secretion and autoantibodies production. PTPN22 gene encodes LYP protein, an inhibitor of T- and B-cell activation. PTPN22 1858C>T polymorphism confers rheumatoid arthritis (RA) susceptibility. Hence, we evaluate the relationship between 1858C>T polymorphism with CD40 and CD154 expression and IFN-γ secretion in RA patients.
PTPN22 1858C>T polymorphism was genotyped in 315 RA patients and 315 control subjects (CS) using PCR-RFLP method. Later, we selected only ten anti-CCP-positive RA patients, naïve to disease-modifying antirheumatic drugs and ten CS, all with known 1858C>T PTPN22 genotype. The CD40 and CD154 membrane expressions were determined by flow cytometry in peripheral B and T cells, correspondingly.
The B cells percentage and mCD40 expression were similar between RA and CS (P > 0.05) and we did not find an association between these variables and the 1858C>T polymorphism. The CD4+ T cells percentage was higher in RA patients than CS (P = 0.003), and in the RA group, the CD4+ T cells percentage and mCD154 expression were higher in the 1858 T allele carriers (P = 0.008 and P = 0.032, respectively). The IFN-γ levels were lower in RA patients carrying the PTPN22 risk allele (P = 0.032).
The PTPN22 1858 T risk allele is associated with increased CD4+ T cells percentage and high mCD154 expression in RA patients, which could favor the pro-inflammatory cytokine release and the establishment of the inflammatory response at the seropositive RA.
CD40是B细胞的共刺激分子，CD154是CD4+T细胞活化的标志物。CD40-CD154相互作用促进促炎细胞因子分泌和自身抗体产生。PTPN22基因编码LYP蛋白，一种T和B细胞活化的抑制剂。PTPN22 1858C>T多态性赋予类风湿关节炎（RA）易感性。因此，我们评估RA患者中1858C>T多态性与CD40和CD154表达以及IFN-γ分泌之间的关系。
采用PCR-RFLP方法对315例RA患者和315例对照受试者（CS）进行PTPN22 1858C>T多态性基因分型。随后，我们仅选择了10例抗CCP阳性的RA患者，这些患者未使用过改善病情的抗风湿药物，以及10例CS，所有患者均已知PTPN22 1858C>T基因型。通过流式细胞术分别在外周血B细胞和T细胞中测定CD40和CD154膜表达。
RA患者和CS之间的B细胞百分比和mCD40表达相似（P>0.05），并且我们未发现这些变量与1858C>T多态性之间存在关联。RA患者的CD4+T细胞百分比高于CS（P=0.003） 在RA组中，1858T等位基因携带者的CD4+T细胞百分比和mCD154表达更高（分别为P=0.008和P=0.032）。携带PTPN22风险等位基因的RA患者的IFN-γ水平较低（P=0.032）。
PTPN22 1858T风险等位基因与RA患者中CD4+T细胞百分比增加和高mCD154表达相关，这可能有利于促炎细胞因子释放以及血清学阳性RA炎症反应的建立。
