Chen Ding, Ireland Sara J, Remington Gina, Alvarez Enrique, Racke Michael K, Greenberg Benjamin, Frohman Elliot M, Monson Nancy L
Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical Center, Dallas, TX 75390.
Department of Neurology, University of Colorado, Aurora, CO 80045.
J Immunol. 2016 Dec 1;197(11):4257-4265. doi: 10.4049/jimmunol.1600782. Epub 2016 Oct 26.
CD40 interacts with CD40L and plays an essential role in immune regulation and homeostasis. Recent research findings, however, support a pathogenic role of CD40 in a number of autoimmune diseases. We previously showed that memory B cells from relapsing-remitting multiple sclerosis (RRMS) patients exhibited enhanced proliferation with CD40 stimulation compared with healthy donors. In this study, we used a multiparameter phosflow approach to analyze the phosphorylation status of NF-κB and three major MAPKs (P38, ERK, and JNK), the essential components of signaling pathways downstream of CD40 engagement in B cells from MS patients. We found that memory and naive B cells from RRMS and secondary progressive MS patients exhibited a significantly elevated level of phosphorylated NF-κB (p-P65) following CD40 stimulation compared with healthy donor controls. Combination therapy with IFN-β-1a (Avonex) and mycophenolate mofetil (Cellcept) modulated the hyperphosphorylation of P65 in B cells of RRMS patients at levels similar to healthy donor controls. Lower disease activity after the combination therapy correlated with the reduced phosphorylation of P65 following CD40 stimulation in treated patients. Additionally, glatiramer acetate treatment also significantly reduced CD40-mediated P65 phosphorylation in RRMS patients, suggesting that reducing CD40-mediated p-P65 induction may be a general mechanism by which some current therapies modulate MS disease.
CD40与CD40配体相互作用，在免疫调节和体内平衡中发挥重要作用。然而，最近的研究结果支持CD40在多种自身免疫性疾病中的致病作用。我们之前表明，与健康供体相比，复发缓解型多发性硬化症（RRMS）患者的记忆B细胞在CD40刺激下增殖增强。在本研究中，我们使用多参数磷酸化流式细胞术方法分析NF-κB和三种主要丝裂原活化蛋白激酶（P38、ERK和JNK）的磷酸化状态，这些是MS患者B细胞中CD40激活下游信号通路的关键组成部分。我们发现，与健康供体对照相比，RRMS和继发进展型MS患者的记忆B细胞和幼稚B细胞在CD40刺激后磷酸化NF-κB（p-P65）水平显著升高。干扰素β-1a（阿沃尼单抗）和霉酚酸酯（骁悉）联合治疗可调节RRMS患者B细胞中P65的过度磷酸化，使其水平与健康供体对照相似。联合治疗后较低的疾病活动度与治疗患者CD40刺激后P65磷酸化减少相关。此外，醋酸格拉替雷治疗也显著降低了RRMS患者中CD40介导的P65磷酸化，这表明减少CD40介导的p-P65诱导可能是目前一些疗法调节MS疾病的普遍机制。
