Distinct contribution of autoreactive B cell Bruton’s tyrosine kinase signaling to neuroinflammation

In multiple sclerosis (MS), autoreactive B cells play a central role in driving CD4 T cell-mediated inflammatory damage to myelin (1). Here we investigated how disrupting Bruton’s tyrosine kinase (BTK) signaling exclusively in B cells shapes the course of experimental autoimmune encephalomyelitis (EAE), a model for MS, through alterations in B cell development and activity. B cell-specific BTK deletion significantly ameliorated both human MOG (hMOG) induced EAE ( p = 0.0087) as well as spontaneous disease in 2D2 + IgH MOG mice ( p = 0.0004). Additionally, MOG-specific cells were found to be more sensitive to loss of BTK than tolerant clones ( p = 0.0002) and production of anti-MOG immunoglobulins was also found to be diminished ( p < 0.004) while overall IgG was unchanged ( p = 0.44). B cells isolated from conditional knockout mice did not upregulate expression of co-stimulatory receptors or MHC II to the same extent as controls when cultured alongside MOG-specific CD4 T cells ( p < 0.005) and were inferior at driving T cell proliferation ( p < 0.0001) in vitro . Lastly, while BTK deletion diminished the proliferative and survival response of B cells following mitogen stimulation, B cell trafficking to the leptomeninges and organization into ectopic lymphoid tissues (ELTs) in 2D2 + IgH MOG mice continued unabated. We identified that BTK signaling regulates several features adopted by autoreactive B cells that contribute to EAE pathogenesis. This study provides mechanistic insights into the therapeutic benefits of BTK inhibitors observed in clinical trials exploring BTK as a therapeutic target in the context of MS. Significance statement Autoreactive B cells contribute to the neuroinflammation that drives multiple sclerosis (MS) and related diseases, yet the molecular mechanisms enabling their pathogenicity remain incompletely understood. This study demonstrates that B cell-specific deletion of Bruton’s tyrosine kinase (BTK) markedly reduces disease severity in two complementary versions of experimental autoimmune encephalomyelitis (EAE), a widely used animal model for MS. Loss of BTK impairs autoreactive B cell survival, antibody production, antigen presentation to encephalitogenic T cells, and T cell activation, while leaving meningeal ectopic lymphoid tissue formation intact. These findings provide direct mechanistic evidence that BTK signaling in B cells promotes neuroinflammatory damage and supports the therapeutic targeting of BTK to limit B cell-driven pathology in MS.