9e1dad322bca9bbbe64a29843eb3dcd5 ijms-26-06898.pdf 27ca04815d9dce12290afdf256f0ee87016e30fb ijms-26-06898.pdf a291fa803e7eb5455f35bfb7b4245b3a5fc45eecea189fc9379f0e7005517c26 ijms-26-06898.pdf Title: Combining CSF and Serum Biomarkers to Differentiate Mechanisms of Disability Worsening in Multiple Sclerosis Subject: The combined use of serum and CSF biomarkers for prognostic stratification in multiple sclerosis (MS) remains underexplored. This multicenter observational study investigated associations between serum neurofilament light chain (sNfL), glial fibrillary acidic protein (sGFAP), and CSF lipid-specific IgM oligoclonal bands (LS-OCMB) with different forms of disability worsening, such as relapse-associated worsening (RAW), active progression independent of relapse activity (aPIRA), and non-active PIRA (naPIRA). A total of 535 patients with MS were included, all sampled within one year of disease onset. Biomarkers were quantified using single-molecule array and immunoblotting techniques, and CSF cell subsets were analyzed by flow cytometry. High sNfL z-scores and LS-OCMB positivity were independently associated with increased risk of RAW and aPIRA, collectively termed inflammatory-associated worsening (IAW), while elevated sGFAP levels predicted naPIRA. Patients with both high sNfL and LS-OCMB positivity had the highest risk of IAW. Among LS-OCMB–positive patients, higher regulatory T cell percentages were associated with lower sNfL levels, suggesting a protective role. Conversely, in LS-OCMB–negative patients, sNfL levels correlated with CSF C3 concentrations. These findings support the complementary role of sNfL, sGFAP, and LS-OCMB in identifying distinct mechanisms of disease worsening and may inform early personalized management strategies in MS. Keywords: multiple sclerosis; neurofilament light chain; glial fibrillary acidic protein; intrathecal IgM synthesis; progression Author: Enric Monreal, José Ignacio Fernández-Velasco, Susana Sainz de la Maza, Mercedes Espiño, Noelia Villarrubia, Ernesto Roldán-Santiago, Yolanda Aladro, Juan Pablo Cuello, Lucía Ayuso-Peralta, Alexander Rodero-Romero, Juan Luís Chico-García, Fernando Rodríguez-Jorge, Ana Quiroga-Varela, Eulalia Rodríguez-Martín, Belén Pilo de la Fuente, Guillermo Martín-Ávila, María Luisa Martínez-Ginés, José Manuel García-Domínguez, Lluïsa Rubio, Sara Llufriu, Manuel Comabella, Xavier Montalban, Gary Álvarez-Bravo, José Luís Veiga-González, Jaime Masjuan, Lucienne Costa-Frossard and Luisa María Villar Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Jul 18 10:37:39 2025 CEST ModDate: Fri Jul 18 10:40:29 2025 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1715850 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PKMTHI+URWPalladioL-Ital Type 1 Custom yes yes yes 124 0 XFDGRF+URWPalladioL-Bold Type 1 Custom yes yes yes 125 0 SSXPJJ+VnURWPalladioL Type 1 Custom yes yes yes 126 0 BVRFOY+URWPalladioL-Roma Type 1 Custom yes yes yes 127 0 SIDQKI+EURM10 Type 1 Builtin yes yes yes 192 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 193 0 HYYHEA+CMBSY10 Type 1 Builtin yes yes yes 244 0 NAHPLP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 209 0 NAHPLN+PalatinoLinotype TrueType WinAnsi yes yes no 210 0 VORBJF+CMSY10 Type 1 Builtin yes yes yes 293 0 NAHPLP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 228 0 NAHPLN+PalatinoLinotype TrueType WinAnsi yes yes no 229 0 CRJHCU+PazoMath-Italic Type 1 Builtin yes yes yes 338 0 NAHPLN+PalatinoLinotype TrueType WinAnsi yes yes no 358 0 NAHPLN+PalatinoLinotype TrueType WinAnsi yes yes no 385 0 NAHPLN+PalatinoLinotype TrueType WinAnsi yes yes no 592 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-05-22 03:08:49 CEST RepresentationInformation: ijms-26-06898.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-05-21 18:33:54 CEST Size: 1715850 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 723 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Results Children: Item: Title: Baseline Characteristics Item: Title: Risk of RAW, Active, and Non-Active PIRA Item: Title: Risk of IAW and naPIRA Across Different Biomarker Groups Item: Title: CSF Cells and Soluble Factors Associated with Biomarkers Item: Title: Discussion Item: Title: Materials and Methods Children: Item: Title: Study Design Item: Title: Data Collection Item: Title: CSF Analyses Item: Title: Soluble Factors Item: Title: MRI Protocols Item: Title: Definitions Item: Title: Statistical Analyses Item: Title: Appendix A Item: Title: References Info: Title: Combining CSF and Serum Biomarkers to Differentiate Mechanisms of Disability Worsening in Multiple Sclerosis Author: Enric Monreal, José Ignacio Fernández-Velasco, Susana Sainz de la Maza, Mercedes Espiño, Noelia Villarrubia, Ernesto Roldán-Santiago, Yolanda Aladro, Juan Pablo Cuello, Lucía Ayuso-Peralta, Alexander Rodero-Romero, Juan Luís Chico-García, Fernando Rodríguez-Jorge, Ana Quiroga-Varela, Eulalia Rodríguez-Martín, Belén Pilo de la Fuente, Guillermo Martín-Ávila, María Luisa Martínez-Ginés, José Manuel García-Domínguez, Lluïsa Rubio, Sara Llufriu, Manuel Comabella, Xavier Montalban, Gary Álvarez-Bravo, José Luís Veiga-González, Jaime Masjuan, Lucienne Costa-Frossard and Luisa María Villar Subject: The combined use of serum and CSF biomarkers for prognostic stratification in multiple sclerosis (MS) remains underexplored. This multicenter observational study investigated associations between serum neurofilament light chain (sNfL), glial fibrillary acidic protein (sGFAP), and CSF lipid-specific IgM oligoclonal bands (LS-OCMB) with different forms of disability worsening, such as relapse-associated worsening (RAW), active progression independent of relapse activity (aPIRA), and non-active PIRA (naPIRA). A total of 535 patients with MS were included, all sampled within one year of disease onset. Biomarkers were quantified using single-molecule array and immunoblotting techniques, and CSF cell subsets were analyzed by flow cytometry. High sNfL z-scores and LS-OCMB positivity were independently associated with increased risk of RAW and aPIRA, collectively termed inflammatory-associated worsening (IAW), while elevated sGFAP levels predicted naPIRA. Patients with both high sNfL and LS-OCMB positivity had the highest risk of IAW. Among LS-OCMB–positive patients, higher regulatory T cell percentages were associated with lower sNfL levels, suggesting a protective role. Conversely, in LS-OCMB–negative patients, sNfL levels correlated with CSF C3 concentrations. These findings support the complementary role of sNfL, sGFAP, and LS-OCMB in identifying distinct mechanisms of disease worsening and may inform early personalized management strategies in MS. 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