Soraphen A effect on B cells post-stroke
Description
Introduction: Immunomodulatory approaches could advance post-stroke therapy. Previous work demonstrated that Soraphen A (SorA) improves functional recovery, in part, by modulating T cell responses. Here, we demonstrate that SorA also directly or indirectly modulates B cell responses. Methods: Male Nur77GFP mice (12–14 weeks) underwent tMCAO. At reperfusion, mice received intraperitoneal LPS or vehicle; SorA or vehicle was given 2 h later and daily thereafter. B cell activation and differentiation were analyzed by flow cytometry in lungs, spleen, blood, and lymph nodes at 16 h, 2, 3, and 7 d, with GFP indicating antigen-specific activation. Results: While both SorA and lipopolysaccharide (LPS) replenished splenic B cell numbers depleted by stroke, their effects on B cell subsets diverge. SorA also reversed LPS-induced reductions in lung B cell counts and decreased plasma cell counts. Both treatments increased B cell activation and modulated MHC-II expression. In contrast to T cells, the mode of B cell activation was not affected by SorA treatment. Effects of LPS and SorA on B cell populations were most distinct in the lungs. Discussion: These findings demonstrate that the beneficial effects of SorA after stroke are not solely mediated by T cell modulation. Further investigation into SorA’s organ-specific impact on different B cell subtypes is crucial to fully elucidate its mechanisms of action.
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Steps to reproduce
-MCAO (45 min), i.p. injection of LPS (40 µg/kg) or vehicle (PBS) 2h at reperfusion, i.p. injection of Soraphen A (SorA; 500 µM in PEG buffer [PBS with 5% PEG400, 2.5% Tween80, and 0.5% ethanol], 5 ml/kg, corresponding to 2.5 µmol/kg) 2h after surgery and daily thereafter. -experimental time points (16 h, 2 d, 3 d, and 7 d post-MCAO): blood collection via cardiac puncture, transcardiac perfusion, collection of spleens, lungs, and lymph nodes (cervical, inguinal, mesenteric) -preparation of single cell suspensions and flow cytometric analysis -data analysis using FlowJo (v. 10.8.1) and excel -statistical analysis using R