Centre for Immunodeficiency, Molecular Immunology Unit, UCL Institute of Child Health, London, United Kingdom.
J Allergy Clin Immunol. 2011 May;127(5):1253-9.e13. doi: 10.1016/j.jaci.2011.02.037. Epub 2011 Mar 31.
Mutations in TNFRSF13B, the gene encoding transmembrane activator and calcium modulator cyclophilin ligand interactor (TACI), are found in 10% of patients with common variable immunodeficiency. However, the most commonly detected mutation is the heterozygous change C104R, which is also found in 0.5% to 1% of healthy subjects. The contribution of the C104R mutation to the B-cell defects observed in patients with common variable immunodeficiency therefore remains unclear.
We sought to define the functional consequences of the C104R mutation on B-cell function.
We performed in vitro studies of TACI C104R expression and signaling. A knock-in mouse with the equivalent mutation murine TACI (mTACI) C76R was generated as a physiologically relevant model of human disease. We examined homozygous and heterozygous C76R mutant mice alongside wild-type littermates and studied specific B-cell lineages and antibody responses to T cell-independent and T cell-dependent challenge.
C104R expression and ligand binding are significantly diminished when the mutant protein is expressed in 293T cells or in patients' cell lines. This leads to defective nuclear factor κB activation, which is proportionally restored by reintroduction of wild-type TACI. Mice heterozygous and homozygous for mTACI C76R exhibit significant B-cell dysfunction with splenomegaly, marginal zone B-cell expansion, diminished immunoglobulin production and serological responses to T cell-independent antigen, and abnormal immunoglobulin synthesis.
These data show that the C104R mutation and its murine equivalent, C76R, can significantly disrupt TACI function, probably through haploinsufficiency. Furthermore, the heterozygous C76R mutation alone is sufficient to disturb B-cell function with lymphoproliferation and immunoglobulin production defects.
TNFRSF13B 基因突变可导致 10%的普通变异性免疫缺陷患者发病,该基因编码跨膜激活物和钙调磷酸酶配体相互作用因子(TACI)。然而,最常见的突变是杂合性改变 C104R,在 0.5%至 1%的健康人群中也可发现该突变。因此,普通变异性免疫缺陷患者中观察到的 B 细胞缺陷与 C104R 突变的关系仍不清楚。
我们旨在明确 C104R 突变对 B 细胞功能的影响。
我们进行了体外 TACI C104R 表达和信号转导研究。构建了携带人源疾病中 C104R 突变的鼠 TACI(mTACI)C76R 基因敲入小鼠,作为研究人类疾病的生理相关模型。我们对 C76R 纯合和杂合突变小鼠及其野生型同窝仔鼠进行了研究,并检测了特定 B 细胞谱系和对 T 细胞非依赖性和 T 细胞依赖性抗原的抗体反应。
当突变蛋白在 293T 细胞或患者细胞系中表达时,C104R 的表达和配体结合明显减少,导致核因子 κB 激活缺陷,野生型 TACI 的重新导入可使缺陷得到比例性恢复。mTACI C76R 杂合和纯合小鼠表现出显著的 B 细胞功能障碍,包括脾肿大、边缘区 B 细胞扩增、免疫球蛋白产生减少以及对 T 细胞非依赖性抗原的血清学反应异常和异常的免疫球蛋白合成。
这些数据表明,C104R 突变及其鼠源等效突变 C76R 可显著破坏 TACI 功能,可能通过杂合性缺失。此外,杂合 C76R 突变本身就足以导致 B 细胞功能障碍,引起淋巴细胞增殖和免疫球蛋白产生缺陷。