Intrathecal antibodies cross-react with EBV BRRF2 and human antigens in multiple sclerosis

Schulte-Frankenfeld, P. M.; Decker, T.; Bünger, I.; Bamberg, S.; Thakar, M.; Morgenlander, W. R.; Schindler, P.; Otto, C.; Sperber, P. S.; Schmitz-Hübsch, T.; Kornau, H.-C.; Schmitz, D.; Jarius, S.; Longbrake, E. E.; Yandamuri, S.; OConnor, K. C.; Schwake, C.; Ayzenberg, I.; Pardo, C. A.; Paul, F.; Calabresi, P. A.; Ruprecht, K.; Larman, H. B.; Kreye, J.

doi:10.64898/2026.08.14.744882

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bioRxiv · 10.64898/2026.08.14.744882

Intrathecal antibodies cross-react with EBV BRRF2 and human antigens in multiple sclerosis

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Abstract

Intrathecal antibody synthesis is a hallmark of multiple sclerosis (MS). Although some intrathecally synthesized antibodies in MS are known to target viral antigens, the spectrum of their antigenic specificities remains incompletely defined. We combined proteome-wide antibody profiling by Phage ImmunoPrecipitation Sequencing (PhIP-Seq) with cross-compartment analytics (MICAR) to study intrathecal antibody synthesis at peptide resolution in paired CSF and serum samples from individuals with MS (n = 40) and non-MS controls (n = 83). While intrathecal antibody responses in MS were polyspecific and included reactivities to various viruses, we identified a subset of individuals with a convergent intrathecal antibody reactivity to a previously described motif within the Epstein-Barr virus (EBV) protein BRRF2 (BRRF2408-415). This motif-directed response showed co-reactivity with multiple CNS-expressed human antigens. In an independent cohort of n = 909 individuals with MS and n = 311 controls, including individuals with NMOSD and MOGAD, serum antibodies to BRRF2408-415 were detected in 8.36% of MS individuals and in 0.64% of non-MS controls, corresponding to an odds ratio for MS of 14.1 (95% CI: 4.4-86.04). Within MS individuals, BRRF2408-415 seropositivity was associated with increased intrathecal IgG synthesis. Cross-reactivity of antibodies to BRRF2408-415 with human targets, including TRIM71 and RTN2, was confirmed by competition ELISA and cell-based assays. Together, these data define an intrathecal EBV BRRF2-linked antibody signature with human target cross-reactivity in a subset of MS individuals. This signature identifies a highly specific serological marker in MS and may support future stratification of the heterogeneous MS spectrum.

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Schulte-Frankenfeld, P. M., Decker, T., Bünger, I., Bamberg, S., Thakar, M., Morgenlander, W. R., Schindler, P., Otto, C., Sperber, P. S., Schmitz-Hübsch, T., Kornau, H.-C., Schmitz, D., Jarius, S., Longbrake, E. E., Yandamuri, S., OConnor, K. C., Schwake, C., Ayzenberg, I., Pardo, C. A., Paul, F., Calabresi, P. A., Ruprecht, K., Larman, H. B., Kreye, J.. 2026-08-20. Intrathecal antibodies cross-react with EBV BRRF2 and human antigens in multiple sclerosis. https://doi.org/10.64898/2026.08.14.744882

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