Table_1_Adaptive and innate immune responses in multiple sclerosis with anti-CD20 therapy: Gene expression and protein profiles.XLSX

BackgroundAnti-CD20 is a highly effective therapy for multiple sclerosis (MS), a disease with multiple abnormalities in function of B and T cells and innate immune cells. Anti-CD20 therapy depletes B cells, which alters antibody production and has diverse effects on B cell immunity. These changes potentially affect immunity beyond B cells in MS.
ObjectiveDetermine if anti-CD20 therapy effects non-B cell, as well as B cell, gene expression, and serum protein levels.
MethodsSamples were collected from 10 healthy controls and from clinically stable relapsing–remitting MS – 10 untreated, 9 interferon-β-treated, and 15 ocrelizumab-treated patients were studied before, and 2  weeks and 6  months after, the first anti-CD20 infusion. Peripheral blood mononuclear cells (PBMC) were analyzed with sensitive, 135,000-transcript RNA expression microarrays, using stringent criteria. Gene expression was compared to 43 MS-relevant serum immune and neurotrophic proteins, using multiplex protein assays.
ResultsAnti-CD20 therapy reduced expression of 413 total genes and 185 B-cell-regulated genes at 2  weeks vs. pre-therapy. Expression of 19 (15%) of these B cell genes returned toward baseline by 6  months, including genes for the B cell activation protein, CD79A, and for immunoglobulin A, D, and G heavy chains. Expression pathways for Th17 and CD4 regulatory T-cell (Treg) development, differentiation, and proliferation also quieted. In contrast, expression increased in Th1 and myeloid cell antiviral, pro-inflammatory, and toll-like receptor (TLR) gene pathways.
ConclusionThese findings have clinical implications. B cell gene expression diminishes 2  weeks after anti-CD20 antibody infusion, but begins to rebound by 6  months. This suggests that the optimum time for vaccination is soon before reinfusion of anti-CD20 therapy. In addition, at 6  months, there is enhanced Th1 cell gene expression and induction of innate immune response genes and TLR expression, which can enhance anti-viral and anti-tumor immunity. This may compensate for diminished B cell gene expression after therapy. These data suggest that anti-CD20 therapy has dynamic effect on B cells and causes a compensatory rise in Th1 and myeloid immunity.

【背景】抗CD20（Anti-CD20）疗法是治疗多发性硬化（multiple sclerosis, MS）的高效手段。MS患者的B细胞（B cell）、T细胞（T cell）及固有免疫细胞（innate immune cells）功能均存在多种异常。抗CD20疗法可耗竭B细胞，进而改变抗体生成过程，并对B细胞免疫功能产生多方面影响。此类变化或可影响MS患者体内非B细胞相关的免疫状态。
【目的】明确抗CD20疗法是否会对MS患者的B细胞及非B细胞的基因表达，以及血清蛋白水平产生影响。
【方法】本研究纳入10名健康对照者，以及临床稳定的复发缓解型MS患者：其中未接受治疗者10例、接受干扰素-β（interferon-β）治疗者9例、接受奥瑞珠单抗（ocrelizumab）治疗者15例。分别在首次抗CD20输注前、输注后2周及6个月采集样本。采用严格的实验标准，通过可检测135000个转录本的高灵敏度RNA表达微阵列（RNA expression microarrays）分析外周血单个核细胞（peripheral blood mononuclear cells, PBMC）的基因表达情况；同时采用多重蛋白检测（multiplex protein assays）技术，将基因表达结果与43种与MS相关的血清免疫蛋白及神经营养蛋白进行比对分析。
【结果】与治疗前相比，抗CD20疗法在输注后2周可使413个总基因及185个B细胞调控基因的表达水平下调。其中19个（占比15%）B细胞相关基因的表达在6个月时恢复至基线水平，包括B细胞活化蛋白CD79A编码基因，以及免疫球蛋白（immunoglobulin）A、D、G重链编码基因。Th17细胞及CD4+调节性T细胞（CD4 regulatory T-cell, Treg）的发育、分化与增殖相关表达通路也被抑制。与之相反，Th1细胞、髓系细胞（myeloid cell）的抗病毒、促炎相关通路以及toll样受体（toll-like receptor, TLR）基因通路的表达水平则出现上调。
【结论】本研究结果具有临床指导意义。抗CD20单抗输注后2周，患者体内B细胞基因表达水平显著降低，但在6个月时开始出现回升。这提示疫苗接种的最佳时机应为抗CD20疗法再次输注前不久。此外，在输注后6个月，患者体内Th1细胞基因表达、固有免疫应答基因及TLR表达均出现上调，可增强抗病毒与抗肿瘤免疫能力，这或许能够弥补疗法实施后B细胞基因表达下调带来的免疫缺陷。本研究数据表明，抗CD20疗法对B细胞具有动态调控作用，并可代偿性上调Th1细胞与髓系细胞的免疫功能。