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Juvenile idiopathic arthritis: lymphocyte activation gene-3 is a central immune receptor in children with oligoarticular subtypes

Abstract

Background

We investigated the role of inhibitory receptors (IRs) and especially lymphocyte activation gene-3 (LAG-3) in the pathogenesis of oligoarticular juvenile idiopathic arthritis (o-JIA).

Methods

Paired samples of synovial fluid (SF) and plasma and peripheral blood (PBMCs) and synovial fluid mononuclear cells (SFMCs) were collected from o-JIA patients along with their clinical data (n = 24). Plasma from healthy controls (n = 14) and paired SF and plasma samples from five non-arthritic juvenile orthopedic patients (n = 5) served as controls. Spontaneously differentiated fibroblast-like synoviocytes (FLSs) from SFMCs were co-cultured with autologous PBMCs/SFMCs and used as ex vivo disease model. Soluble levels and cellular expressions of IRs together with their functional properties in the ex vivo model were analyzed.

Results

In patients with o-JIA, soluble levels of LAG-3 and expression of LAG-3 and T cell immunoglobulin mucin03 (TIM-3) on CD3+CD4+CD45RO+ T cells were increased, especially in SF. Major histocompatibility complex (MHC) class II expression was induced on FLSs when these were co-cultured with autologous PBMCs/SFMCs, together with an increased monocyte chemoattractant protein-1 (MCP-1) production. In PBMC and FLS + PBMC co-cultures, neutralizing antibodies to IRs were added. Only anti-LAG-3 antibodies significantly increased MCP-1 secretion. The addition of agonistic LAG-3 antibody resulted in decreased effector cytokine secretion.

Conclusions

This is the first report comparing the effects of different IRs in o-JIA and suggests that LAG-3 might contribute to the pathogenesis of this disease.

Impact

  • This is the first study addressing the role of different co-IRs in o-JIA.

  • We showed that LAG-3 and TIM-3 seem more important in juvenile arthritis in contrast to adult rheumatoid arthritis, where cytotoxic T-lymphocyte-associated antigen-4 and programmed cell death-1 were reported to be more important.

  • We designed an ex vivo disease model for o-JIA, examined the effects of co-IRs in this model, and demonstrated that they might contribute to the pathogenesis of the disease.

  • LAG-3 might play a role in o-JIA pathogenesis and might be a potential therapeutic option for o-JIA patients.

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Fig. 1: Soluble (s) levels of PD-1, LAG-3, and TIM-3 in plasma and SF in oligoarticular JIA, non-arthritis juvenile orthopedic controls, and healthy controls.
Fig. 2: Surface expressions of IRs, CTLA-4, PD-1, LAG-3, and TIM-3, in JIA PBMC and SFMC (n = 11).
Fig. 3: MHC class II expressions on CD45-PDPN + FLS (n = 4).
Fig. 4: MCP-1 levels in the co-culture supernatants of ex vivo arthritis model (n = 6).
Fig. 5: Different cytokine levels in the FLS + PBMC co-cultures (n = 5) after 48 h.

Data availability

All data generated or analyzed during this study are included in this paper and in its Supplementary information files.

References

  1. Prakken, B., Albani, S. & Martini, A. Juvenile idiopathic arthritis. Lancet 377, 2138–2149 (2011).

    Article  Google Scholar 

  2. Petty, R. E. et al. International League of Associations for Rheumatology classification of juvenile idiopathic arthritis: second revision, Edmonton, 2001. J. Rheumatol. 31, 390–392 (2004).

    PubMed  Google Scholar 

  3. Ozen, S. et al. Prevalence of juvenile chronic arthritis and familial Mediterranean fever in Turkey: a field study. J. Rheumatol. 25, 2445–2449 (1998).

    CAS  PubMed  Google Scholar 

  4. Cimaz, R., Marino, A. & Martini, A. How I treat juvenile idiopathic arthritis: a state of the art review. Autoimmun. Rev. 16, 1008–1015 (2017).

    Article  Google Scholar 

  5. Hinks, A. et al. Fine-mapping the MHC locus in juvenile idiopathic arthritis (JIA) reveals genetic heterogeneity corresponding to distinct adult inflammatory arthritic diseases. Ann. Rheum. Dis. 76, 765–772 (2017).

    CAS  Article  Google Scholar 

  6. McIntosh, L. A. et al. Genome-wide association meta-analysis reveals novel juvenile idiopathic arthritis susceptibility loci. Arthritis Rheumatol. 69, 2222–2232 (2017).

    CAS  Article  Google Scholar 

  7. Hinks, A. et al. Dense genotyping of immune-related disease regions identifies 14 new susceptibility loci for juvenile idiopathic arthritis. Nat. Genet. 45, 664–669 (2013).

    CAS  Article  Google Scholar 

  8. Amariglio, N. et al. T-cell compartment in synovial fluid of pediatric patients with JIA correlates with disease phenotype. J. Clin. Immunol. 31, 1021–1028 (2011).

    Article  Google Scholar 

  9. Wedderburn, L. R., Patel, A., Varsani, H. & Woo, P. Divergence in the degree of clonal expansions in inflammatory T cell subpopulations mirrors HLA-associated risk alleles in genetically and clinically distinct subtypes of childhood arthritis. Int. Immunol. 13, 1541–1550 (2001).

    CAS  Article  Google Scholar 

  10. Nussing, S., Trapani, J. A. & Parish, I. A. Revisiting T cell tolerance as a checkpoint target for cancer immunotherapy. Front. Immunol. 11, 589641 (2020).

    CAS  Article  Google Scholar 

  11. Zhang, Q. & Vignali, D. A. Co-stimulatory and co-inhibitory pathways in autoimmunity. Immunity 44, 1034–1051 (2016).

    CAS  Article  Google Scholar 

  12. Schnell, A., Bod, L., Madi, A. & Kuchroo, V. K. The yin and yang of co-inhibitory receptors: toward anti-tumor immunity without autoimmunity. Cell Res. 30, 285–299 (2020).

    CAS  Article  Google Scholar 

  13. Wherry, E. J. & Kurachi, M. Molecular and cellular insights into T cell exhaustion. Nat. Rev. Immunol. 15, 486–499 (2015).

    CAS  Article  Google Scholar 

  14. McKinney, E. F. & Smith, K. G. C. Metabolic exhaustion in infection, cancer and autoimmunity. Nat. Immunol. 19, 213–221 (2018).

    CAS  Article  Google Scholar 

  15. Greisen, S. R. et al. Increased soluble programmed death-1 (sPD-1) is associated with disease activity and radiographic progression in early rheumatoid arthritis. Scand. J. Rheumatol. 43, 101–108 (2014).

    CAS  Article  Google Scholar 

  16. Moseley, K. F. et al. Immune-related adverse events with immune checkpoint inhibitors affecting the skeleton: a seminal case series. J. Immunother. Cancer 6, 104 (2018).

    Article  Google Scholar 

  17. Mizoguchi, F. et al. Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis. Nat. Commun. 9, 789 (2018).

    Article  Google Scholar 

  18. Dakin, S. G. et al. Pathogenic stromal cells as therapeutic targets in joint inflammation. Nat. Rev. Rheumatol. 14, 714–726 (2018).

    Article  Google Scholar 

  19. Greisen, S. R. et al. Spontaneous generation of functional osteoclasts from synovial fluid mononuclear cells as a model of inflammatory osteoclastogenesis. APMIS 123, 779–786 (2015).

    CAS  Article  Google Scholar 

  20. Schmidt, E. M. et al. Selective blockade of tumor necrosis factor receptor I inhibits proinflammatory cytokine and chemokine production in human rheumatoid arthritis synovial membrane cell cultures. Arthritis Rheum. 65, 2262–2273 (2013).

    CAS  Article  Google Scholar 

  21. Kragstrup, T. W., Vorup-Jensen, T., Deleuran, B. & Hvid, M. A simple set of validation steps identifies and removes false results in a sandwich enzyme-linked immunosorbent assay caused by anti-animal IgG antibodies in plasma from arthritis patients. SpringerPlus 2, 263 (2013).

    Article  Google Scholar 

  22. Qu, D. & Green, M. Folding and assembly of a human MHC class II molecule in a cell-free system. DNA Cell Biol. 14, 741–751 (1995).

    CAS  Article  Google Scholar 

  23. Huard, B., Gaulard, P., Faure, F., Hercend, T. & Triebel, F. Cellular expression and tissue distribution of the human LAG-3-encoded protein, an MHC class II ligand. Immunogenetics 39, 213–217 (1994).

    CAS  Article  Google Scholar 

  24. Kisielow, M., Kisielow, J., Capoferri-Sollami, G. & Karjalainen, K. Expression of lymphocyte activation gene 3 (LAG-3) on B cells is induced by T cells. Eur. J. Immunol. 35, 2081–2088 (2005).

    CAS  Article  Google Scholar 

  25. Workman, C. J. et al. LAG-3 regulates plasmacytoid dendritic cell homeostasis. J. Immunol. 182, 1885–1891 (2009).

    CAS  Article  Google Scholar 

  26. Ellis, J. et al. Depletion of LAG-3(+) T cells translated to pharmacology and improvement in psoriasis disease activity: a Phase I Randomized Study of mAb GSK2831781. Clin. Pharmacol. Ther. 109, 1293–1303 (2021).

  27. Petrelli, A. et al. PD-1+CD8+ T cells are clonally expanding effectors in human chronic inflammation. J. Clin. Invest. 128, 4669–4681 (2018).

    Article  Google Scholar 

  28. Ronaghy, A. et al. Vaccination leads to an aberrant FOXP3 T-cell response in non-remitting juvenile idiopathic arthritis. Ann. Rheum. Dis. 70, 2037–2043 (2011).

    CAS  Article  Google Scholar 

  29. Frenz, T. et al. CD4(+) T cells in patients with chronic inflammatory rheumatic disorders show distinct levels of exhaustion. J. Allergy Clin. Immunol. 138, 586–589 (2016).

    CAS  Article  Google Scholar 

  30. Huang, C. T. et al. Role of LAG-3 in regulatory T cells. Immunity 21, 503–513 (2004).

    CAS  Article  Google Scholar 

  31. Henderson, L. A. et al. Next-generation sequencing reveals restriction and clonotypic expansion of Treg cells in juvenile idiopathic arthritis. Arthritis Rheumatol. 68, 1758–1768 (2016).

    CAS  Article  Google Scholar 

  32. Rosenzwajg, M. et al. Immunological and clinical effects of low-dose interleukin-2 across 11 autoimmune diseases in a single, open clinical trial. Ann. Rheum. Dis. 78, 209–217 (2019).

    CAS  Article  Google Scholar 

  33. Ruperto, N. et al. Long-term safety and efficacy of abatacept in children with juvenile idiopathic arthritis. Arthritis Rheumatol. 62, 1792–1802 (2010).

    CAS  Article  Google Scholar 

  34. Brunner, H. et al. Subcutaneous abatacept in patients with polyarticular-course juvenile idiopathic arthritis: results from a Phase III Open-Label Study. Arthritis Rheumatol. 70, 144–1154 (2018).

    Article  Google Scholar 

  35. Maggi, L. et al. Immunosuppressive activity of abatacept on circulating T helper lymphocytes from juvenile idiopathic arthritis patients. Int. Arch. Allergy Immunol. 171, 45–53 (2016).

    CAS  Article  Google Scholar 

  36. Fleury, M. et al. Increased expression and modulated regulatory activity of coinhibitory receptors PD-1, TIGIT, and TIM-3 in lymphocytes from patients with systemic sclerosis. Arthritis Rheumatol. 70, 566–577 (2018).

    CAS  Article  Google Scholar 

  37. Postow, M. A., Sidlow, R. & Hellmann, M. D. Immune-related adverse events associated with immune checkpoint blockade. N. Engl. J. Med. 378, 158–168 (2018).

    CAS  Article  Google Scholar 

  38. Boutros, C. et al. Safety profiles of anti-CTLA-4 and anti-PD-1 antibodies alone and in combination. Nat. Rev. Clin. Oncol. 13, 473–486 (2016).

    CAS  Article  Google Scholar 

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Acknowledgements

We would like to thank Karin Skovgaard Sørensen for her technical assistance during this study; to Aarhus University FACS core facility for their advice on the flow cytometry analyses, to Bahar Guclu, the patient/parent research partner, for her contribution. LAG-3 agonist (IMP761) was generously provided by Immutep and Dr. Triebel, CSO and CMO—per clause 10.2 of the MTA. This study was supported by a research grant from FOREUM Foundation for Research in Rheumatology. This study was also supported by the Danish Rheumatoid Association, Aarhus University, The Scientific and Technological Research Council of Turkey (TUBITAK), and the Hacettepe University Scientific Research Unit.

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Authors

Contributions

E.S., S.O., and B.D. designed the study, collected the data, contributed to the data interpretation, and wrote the manuscript; E.S., S.D., E.T., Y.B., and S.O. collected patients’ data and samples, contributed the data interpretation, revised the manuscript; E.S., M.A., M.A.N., C.S., M.H., S.G., and B.D. designed and performed the experiments, contributed the data interpretation, and revised the manuscript. All authors made substantial contributions to either the conception and design of this study or to the generation, analysis, and/or interpretation of data. They agree to be accountable for the integrity of the work herein. All authors reviewed the manuscript and approved the submitted version.

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Correspondence to Erdal Sag.

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The authors declare no competing interests.

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Written informed written consent for publication was obtained from the parents and patients.

Ethical approval and consent to participate

This study was approved by the Hacettepe University Ethics Committee for Non-Interventional Clinical Trials (GO 18/277-35), and informed written consent to participate in the study was obtained from the parents and patients.

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Sag, E., Demir, S., Aspari, M. et al. Juvenile idiopathic arthritis: lymphocyte activation gene-3 is a central immune receptor in children with oligoarticular subtypes. Pediatr Res 90, 744–751 (2021). https://doi.org/10.1038/s41390-021-01588-2

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