Department of Clinical Immunology, Sapienza University of Rome, Rome, Italy.
Department of Molecular Medicine, Sapienza University of Rome, Rome, Italy.
J Allergy Clin Immunol. 2014 Aug;134(2):401-10. doi: 10.1016/j.jaci.2014.03.017. Epub 2014 May 1.
Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by B-cell dysfunction and, in a subgroup, by expansion of CD21(low) B cells. The CD21(low) B cells display defects in early B-cell receptor (BCR) signaling resembling those of anergic B cells.
We sought to investigate whether B cells from patients with CVID, like anergic B cells, have defects in extracellular signal-regulated kinase (ERK) phosphorylation and in endocytic trafficking of the BCR.
Using flow cytometry, we evaluated phosphorylated ERK (pERK) expression and internalization of cross-linked BCR in B-cell subsets. The localization of internalized BCR to lysosome-associated membrane protein 1-positive late endosomes was evaluated with confocal microscopy.
Constitutive pERK levels were increased in naive and IgM(+) memory B cells of patients with CVID compared with those of healthy donors, whereas the pERK increment induced by BCR cross-linking was relatively reduced. Intravenous immunoglobulin administration enhanced these anomalies, but they appeared to be intrinsic to B cells from patients with CVID. Cross-linking-induced BCR endocytosis was decreased in the IgM(+) memory B cells, especially in those with a CD21(low) phenotype, but not in the naive B cells of patients with CVID with CD21(low) expansion. Internalized BCR localized normally to late endosomes. Pharmacologic inhibition of ERK phosphorylation suppressed BCR endocytosis in B cells of healthy patients and those with CVID.
The B cells of patients with CVID with CD21(low) B-cell expansion resemble anergic B cells based on high constitutive pERK expression. The IgM(+) memory B cells of these patients, especially those that are CD21(low), have a defect in BCR endocytosis seemingly caused by dysregulated ERK signaling.
普通变异型免疫缺陷(CVID)是一种以 B 细胞功能障碍为特征的异质性疾病,在亚组中,以 CD21(low)B 细胞扩增为特征。CD21(low)B 细胞表现出类似于无反应性 B 细胞的早期 B 细胞受体(BCR)信号传导缺陷。
我们试图研究 CVID 患者的 B 细胞是否像无反应性 B 细胞一样,在细胞外信号调节激酶(ERK)磷酸化和 BCR 的内吞作用方面存在缺陷。
我们使用流式细胞术评估 B 细胞亚群中磷酸化 ERK(pERK)表达和交联 BCR 的内化。通过共聚焦显微镜评估内化的 BCR 到溶酶体相关膜蛋白 1 阳性晚期内体的定位。
与健康供体相比,CVID 患者的幼稚和 IgM(+)记忆 B 细胞中的组成型 pERK 水平增加,而 BCR 交联诱导的 pERK 增加相对减少。静脉注射免疫球蛋白给药增强了这些异常,但它们似乎是 CVID 患者 B 细胞的固有特性。交联诱导的 BCR 内吞作用在 IgM(+)记忆 B 细胞中减少,特别是在具有 CD21(low)表型的细胞中,但在 CVID 患者的幼稚 B 细胞中没有减少,这些患者的 CD21(low)扩展。内化的 BCR 正常定位于晚期内体。ERK 磷酸化的药理学抑制抑制了健康患者和 CVID 患者的 B 细胞中的 BCR 内吞作用。
具有 CD21(low)B 细胞扩增的 CVID 患者的 B 细胞基于高组成型 pERK 表达类似于无反应性 B 细胞。这些患者的 IgM(+)记忆 B 细胞,特别是那些 CD21(low)的细胞,在 BCR 内吞作用中存在缺陷,这似乎是由 ERK 信号传导失调引起的。