α4-integrin deficiency in B cells does not affect disease in a T-cell–mediated EAE disease model

Abstract

ObjectiveThe goal of this study was to investigate the role of CD 19+ B cells within the brain and spinal cord during CNS autoimmunity in a peptide-induced, primarily T-cell–mediated experimental autoimmune encephalomyelitis (EAE) model of MS. We hypothesized that CD19+ B cells outside the CNS drive inflammation in EAE.MethodsWe generated CD19.Cre+/− α4-integrinfl/fl mice. EAE was induced by active immunization with myelin oligodendrocyte glycoprotein peptide (MOGp35-55). Multiparameter flow cytometry was used to phenotype leukocyte subsets in primary and secondary lymphoid organs and the CNS. Serum cytokine levels and Ig levels were assessed by bead array. B-cell adoptive transfer was used to determine the compartment-specific pathogenic role of antigen-specific and non–antigen-specific B cells.ResultsA genetic ablation of α4-integrin in CD19+/− B cells significantly reduced the number of CD19+ B cells in the CNS but does not affect EAE disease activity in active MOGp35-55-induced disease. The composition of B-cell subsets in the brain, primary lymphoid organs, and secondary lymphoid organs of CD19.Cre+/− α4-integrinfl/fl mice was unchanged during MOGp35-55-induced EAE. Adoptive transfer of purified CD19+ B cells from CD19.Cre+/− α4-integrinfl/fl mice or C57BL/6 wild-type (WT) control mice immunized with recombinant rMOG1-125 or ovalbumin323-339 into MOGp35-55-immunized CD19.Cre+/− α4-integrinfl/fl mice caused worse clinical EAE than was observed in MOGp35-55-immunized C57BL/6 WT control mice that did not receive adoptively transferred CD19+ B cells.ConclusionsObservations made in CD19.Cre+/− α4-integrinfl/fl mice in active MOGp35-55-induced EAE suggest a compartment-specific pathogenic role of CD19+ B cells mostly outside of the CNS that is not necessarily antigen specific.

CD11c+CD88+CD317+myeloid cells are critical mediators of persistent CNS autoimmunity

“…Cells from lymph nodes and spleen were isolated by pressing through a 70-µm nylon mesh cell retainer as described previously ( 8 – 10 ). Cells were treated with red blood cell (RBC) lysis buffer (Sigma-Aldrich), washed twice with cold PBS, and resuspended in PBS for counting by hemocytometer.…”

Skin-Homing Regulatory B Cells Required for Suppression of Cutaneous Inflammation

“…In support of potential skin-resident B cells are recent studies demonstrating non-recirculatory spleen-resident T-bet hi B cells (Johnson et al, 2020). Several studies reported that IL-10 + Bregs exert their function by acting in secondary lymphoid organs (Hussain et al, 2019, Matsushita et al, 2008, and constitutive lack of α4β1integrin in B cells has been linked to a reduction of IL-10 + Bregs in spleen and exacerbated CNS inflammation in experimental autoimmune encephalitis (Glatigny et al, 2016). However, in our study, increased inflammation in both CHS-like and psoriasiform skin inflammation was paralleled by a decrease in cutaneous IL-10 + Breg numbers and a concomitant increase of their counterparts in spleen.…”

CD11c+CD88+CD317+myeloid cells are critical mediators of persistent CNS autoimmunity

“…Cells from lymph nodes and spleen were isolated by pressing through a 70 µm nylon mesh cell retainer as described [8][9][10] . Cells were treated with red blood cell (RBC) lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), washed twice with cold phosphate buffered saline (PBS), and re-suspended in PBS for counting by hemocytometer.…”