Persistent Classical and Atypical Memory B Cells Underlie Heterogeneous Vaccine Responses in Ocrelizumab-Treated Multiple Sclerosis

Curtin, R.; Velmurugu, Y.; Dibba, F.; Hao, Y.; Sreenivasaiah, C.; Khodadadi-Jamayran, A.; Nyovanie, S. T.; Kim, A.; Samanovic-Golden, M.; Mulligan, M. J.; Priest, J.; Cabatingan, M.; Winger, R.; Patskovsky, Y.; Kister, I.; Silverman, G. J.; Krogsgaard, M.

doi:10.1101/2025.11.03.686372

bioRxiv · 10.1101/2025.11.03.686372

Persistent Classical and Atypical Memory B Cells Underlie Heterogeneous Vaccine Responses in Ocrelizumab-Treated Multiple Sclerosis

Curtin, R.·Velmurugu, Y.·Dibba, F.·Hao, Y.·Sreenivasaiah, C.·Khodadadi-Jamayran, A.·Nyovanie, S. T.·Kim, A.·Samanovic-Golden, M.·Mulligan, M. J.·Priest, J.·Cabatingan, M.·Winger, R.·Patskovsky, Y.·Kister, I.·Silverman, G. J.·Krogsgaard, M.

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Abstract

Background and ObjectivesPatients with multiple sclerosis (pwMS) treated with ocrelizumab (OCR), a B-cell-depleting therapy, exhibit heterogeneous humoral responses to SARS-CoV-2 mRNA vaccination. The mechanisms underlying this variability remain incompletely understood. We performed a longitudinal analysis of B-cell subset dynamics and antigen-specific T cell responses in OCR-treated pwMS and healthy controls to determine how immune cell composition, timing of OCR infusion, and lymphocyte dynamics influence humoral response outcomes. MethodsBased on post-vaccination anti-Spike IgG titers measured by multiplex bead immunoassay, pwMS were categorized as super-responders (SR), responders (R), or non-responders (NR). A 35-marker spectral flow cytometry panel was used to characterize T- and B-cell subsets longitudinally, at baseline and following stimulation with a SARS-CoV-2 peptide pool. ResultsCD4+ and CD8+ T cell populations were preserved across OCR-treated pwMS, and SARS-CoV-2-specific T cells remained detectable for more than 6 months after vaccination. In contrast, residual B-cell subset composition distinguished responders from non-responders. DN2-like B cells (CD19+CD27-IgD-T-bet+CD11c+CXCR5-) persisted despite repeated OCR infusions and were enriched in SR compared with NR. Repletion of mature naive B cells in peripheral blood correlated with time since last OCR infusion and with stronger humoral immune responses. ConclusionsB-cell subsets that resist OCR-mediated depletion may contribute to vaccine responsiveness in OCR-treated pwMS. Altered repletion kinetics of naive B cell subsets in non-responders suggest that specific B-cell populations may serve as predictive biomarkers of vaccine-induced humoral immunity, despite preserved T-cell responses.

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Curtin, R., Velmurugu, Y., Dibba, F., Hao, Y., Sreenivasaiah, C., Khodadadi-Jamayran, A., Nyovanie, S. T., Kim, A., Samanovic-Golden, M., Mulligan, M. J., Priest, J., Cabatingan, M., Winger, R., Patskovsky, Y., Kister, I., Silverman, G. J., Krogsgaard, M.. 2025-11-04. Persistent Classical and Atypical Memory B Cells Underlie Heterogeneous Vaccine Responses in Ocrelizumab-Treated Multiple Sclerosis. https://doi.org/10.1101/2025.11.03.686372

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