Serum Galectin-9 and TIM-3 Expression in Relapse-Remitting Multiple Sclerosis: Assessment of Immune Checkpoint Biomarkers
Abstract
Objective: An immune checkpoint molecule, T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), is an important effector of T cell exhaustion and tolerance by interacting with its ligand, Galectin-9 (Gal-9). This study investigated whether serum TIM-3/Gal-9 levels could reflect immune regulatory dysfunction in relapsing-remitting multiple sclerosis (RRMS).
Material and Methods: Serum TIM-3 and Gal-9 were evaluated intwo main groups of the study (RRMS patients and control groups) with enzyme-linked immunosorbent assay (ELISA). For the evaluation of diagnostic utility, statistical studies were performed using the Mann-Whitney U test, Spearman’s correlation, and receiver operating characteristic (ROC) curve analysis.
Results: TIM-3 levels were substantially higher in patients than in controls. Gal-9 levels revealed no significant difference, but were markedly heterogeneous in patients. There was a significant positive correlation between study markers in both groups. ROC analysis revealed that TIM-3 had modest diagnostic efficacy, but Gal-9 lacked discriminatory value. A subgroup analysis revealed no significant difference in biomarker levels between individuals on disease-modifying medications and those not on treatment.
Conclusion: This study represents the first evidence that serum TIM-3 is considerably higher in RRMS, indicating immune checkpoint dysregulation, which is fundamental to MS pathogenesis. Although Gal-9 did not have direct diagnostic value, its strong, significant correlation with TIM-3 suggests dysregulation of immune tolerance mechanisms. These findings require validation in large cohorts with clinical correlation, but identify TIM-3 as a candidate immunological exhaustion biomarker, and Gal-9/TIM-3 axis as a potential therapeutic target.
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Rodríguez Murúa S, Farez MF, Quintana FJ. The immune response in multiple sclerosis. Annu Rev Pathol 2022;17:121–39.
Del Negro I, Pez S, Gigli GL, Valente M. Disease activity and progression in multiple sclerosis: new evidences and future perspectives. J Clin Med 2022;11:6643.
Ghasemi N, Razavi S, Nikzad E. Multiple sclerosis: pathogenesis, symptoms, diagnoses and cell-based therapy. Cell J 2017;19:1-10.
Khan Z, Mehan S, Gupta GD, Narula AS. Immune system dysregulation in the progression of multiple sclerosis: molecular insights and therapeutic implications. Neuroscience 2024;548:9-26.
Rezazadeh-Gavgani E, Majidazar R, Lotfinejad P, Kazemi T, Shamekh A. Immune checkpoint molecules: a review on pathways and immunotherapy implications. Immun Inflamm Dis 2025;13:e70196.
Chen H, Zha J, Tang R, Chen G. T-cell immunoglobulin and mucin-domain containing-3 (TIM-3): solving a key puzzle in autoimmune diseases. Int Immunopharmacol 2023;121:110418.
Hastings WD, Anderson DE, Kassam N, Koguchi K, Greenfield EA, Kent SC, et al. TIM-3 is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines. Eur J Immunol 2009;39:2492-501.
Tang R, Rangachari M, Kuchroo VK. Tim-3: a co-receptor with diverse roles in T cell exhaustion and tolerance. Semin Immunol 2019;42:101302.
Zhang M, Liu C, Li Y, Li H, Zhang W, Liu J, et al. Galectin-9 in cancer therapy: from immune checkpoint ligand to promising therapeutic target. Front Cell Dev Biol 2024;11:1332205.
Yang R, Sun L, Li CF, Li CF, Zhao Y, Li X, et al. Galectin-9 interacts with PD-1 and TIM-3 to regulate T cell death and is a target for cancer immunotherapy. Nat Commun 2021;12:832.
Joller N, Anderson AC, Kuchroo VK. LAG-3, TIM-3, and TIGIT: distinct functions in immune regulation. Immunity 2024;57:206-22.
Steelman AJ, Li J. Astrocyte galectin-9 potentiates microglial TNF secretion. J Neuroinflammation 2014;11:144.
Tang L, Li G, Zheng Y, Hou C, Gao Y, Hao Y, et al. Tim-3 relieves experimental autoimmune encephalomyelitis by suppressing MHC-II. Front Immunol 2022;12:770402.
Chertcoff A, Schneider R, Azevedo CJ, Sicotte N, Oh J. Recent advances in diagnostic, prognostic, and disease-monitoring biomarkers in multiple sclerosis. Neurol Clin 2024;42:15-38.
Yang W, Liu C, Li Z, Li Y, Wang Y, Zhang Y. Multi-omic biomarkers associated with multiple sclerosis: from Mendelian randomization to drug prediction. Sci Rep 2025;15:9421.
Confavreux C, Compston A. The natural history of multiple sclerosis. In: Compston A, editor. McAlpine’s Multiple Sclerosis. 4th ed. Philadelphia: Churchill Livingstone Elsevier; 2006;p. 183-272.
Chiba M, Kojima R, Tsuchiya N, Iizuka J, Ishibashi K, Amano H, et al. Soluble TIM-3 levels are increased in patients with diffuse cutaneous systemic sclerosis. J Dermatol Sci 2017;86:4–10.
Kojima R, Ohno T, Iikura M, Niki T, Hirashima M, Iwaya K, et al. Galectin-9 enhances cytokine secretion but suppresses survival and degranulation in a human mast cell line. PLoS One 2014;9:e86106.
So CY, Li Y, Chow KT. New insights on galectin-9 expression in cancer prognosis: an updated systematic review and meta-analysis. PLoS One 2025;20:e0320441.
Tang R, Rangachari M, Kuchroo VK. Tim-3: a co-receptor with diverse roles in T cell exhaustion and tolerance. Semin Immunol 2019;42:101302.
Minnie SA, Kuns RD, Gartlan KH, Zhang P, Wilkinson AN, Samson L, et al. TIM-3+ CD8 T cells with a terminally exhausted phenotype retain functional capacity in hematological malignancies. Sci Immunol 2024;9:eadg1094.
Hu H, Li H, Li R, Fu R, Xing Z, Zhang X, et al. Re-establishing immune tolerance in multiple sclerosis: focusing on novel mechanisms of mesenchymal stem cell regulation of Th17/Treg balance. J Transl Med 2024;22:663.
Feng X, Feng J. Clinical significance of Tim3-positive T cell subsets in patients with multiple sclerosis. J Clin Neurosci 2016;34:193-7.
Li G, Huang R, Fan W, Wang D, Wu F, Zeng F, et al. Galectin-9/TIM-3 as a key regulator of immune response in gliomas with chromosome 1p/19q codeletion. Front Immunol 2021;12:800928.
Weinstock-Guttman B, Sormani MP, Repovic P. Predicting long-term disability in multiple sclerosis: a narrative review of current evidence and future directions. Int J MS Care 2022;24:184-8.
Hung M, Bounsanga J, Voss MW. Interpretation of correlations in clinical research. Postgrad Med 2017;129:902-6.
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