Yu Jong W, Hoffman Sandy, Beal Allison M, Dykon Angela, Ringenberg Michael A, Hughes Anna C, Dare Lauren, Anderson Amber D, Finger Joshua, Kasparcova Viera, Rickard David, Berger Scott B, Ramanjulu Joshi, Emery John G, Gough Peter J, Bertin John, Foley Kevin P
Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, United States of America.
Department of Safety Assessment, GlaxoSmithKline, King of Prussia, United States of America.
PLoS One. 2015 May 12;10(5):e0127083. doi: 10.1371/journal.pone.0127083. eCollection 2015.
CARMA-BCL10-MALT1 signalosomes play important roles in antigen receptor signaling and other pathways. Previous studies have suggested that as part of this complex, MALT1 functions as both a scaffolding protein to activate NF-κB through recruitment of ubiquitin ligases, and as a protease to cleave and inactivate downstream inhibitory signaling proteins. However, our understanding of the relative importance of these two distinct MALT1 activities has been hampered by a lack of selective MALT1 protease inhibitors with suitable pharmacologic properties. To fully investigate the role of MALT1 protease activity, we generated mice homozygous for a protease-dead mutation in MALT1. We found that some, but not all, MALT1 functions in immune cells were dependent upon its protease activity. Protease-dead mice had defects in the generation of splenic marginal zone and peritoneal B1 B cells. CD4+ and CD8+ T cells displayed decreased T cell receptor-stimulated proliferation and IL-2 production while B cell receptor-stimulated proliferation was partially dependent on protease activity. In dendritic cells, stimulation of cytokine production through the Dectin-1, Dectin-2, and Mincle C-type lectin receptors was also found to be partially dependent upon protease activity. In vivo, protease-dead mice had reduced basal immunoglobulin levels, and showed defective responses to immunization with T-dependent and T-independent antigens. Surprisingly, despite these decreased responses, MALT1 protease-dead mice, but not MALT1 null mice, developed mixed inflammatory cell infiltrates in multiple organs, suggesting MALT1 protease activity plays a role in immune homeostasis. These findings highlight the importance of MALT1 protease activity in multiple immune cell types, and in integrating immune responses in vivo.
CARMA-BCL10-MALT1信号小体在抗原受体信号传导及其他信号通路中发挥重要作用。先前的研究表明,作为该复合体的一部分,MALT1既作为一种支架蛋白,通过募集泛素连接酶来激活核因子κB,又作为一种蛋白酶,裂解并失活下游抑制性信号蛋白。然而,由于缺乏具有合适药理特性的选择性MALT1蛋白酶抑制剂,我们对这两种不同的MALT1活性的相对重要性的理解受到了阻碍。为了全面研究MALT1蛋白酶活性的作用,我们构建了MALT1蛋白酶死亡突变的纯合小鼠。我们发现,免疫细胞中部分而非全部的MALT1功能依赖于其蛋白酶活性。蛋白酶死亡小鼠在脾边缘区和腹膜B1 B细胞的生成方面存在缺陷。CD4+和CD8+ T细胞在受到T细胞受体刺激时增殖和白细胞介素-2产生减少,而B细胞受体刺激的增殖部分依赖于蛋白酶活性。在树突状细胞中,通过Dectin-1、Dectin-2和Mincle C型凝集素受体刺激细胞因子产生也部分依赖于蛋白酶活性。在体内,蛋白酶死亡小鼠的基础免疫球蛋白水平降低,并且对依赖T细胞和不依赖T细胞的抗原免疫反应存在缺陷。令人惊讶的是,尽管这些反应有所降低,但MALT1蛋白酶死亡小鼠而非MALT1基因敲除小鼠在多个器官出现混合性炎性细胞浸润,这表明MALT1蛋白酶活性在免疫稳态中发挥作用。这些发现突出了MALT1蛋白酶活性在多种免疫细胞类型以及整合体内免疫反应中的重要性。