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01.12.2018 | Research article | Ausgabe 1/2018 Open Access

Arthritis Research & Therapy 1/2018

Synovial IL-9 facilitates neutrophil survival, function and differentiation of Th17 cells in rheumatoid arthritis

Arthritis Research & Therapy > Ausgabe 1/2018
Kaustav Chowdhury, Uma Kumar, Soumabha Das, Jaydeep Chaudhuri, Prabin Kumar, Maumita Kanjilal, Parashar Ghosh, Geetabali Sircar, Ravi Kiran Basyal, Uma Kanga, Santu Bandyopadhaya, Dipendra Kumar Mitra
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Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s13075-017-1505-8) contains supplementary material, which is available to authorized users.



Role of Th9 cells and interleukin-9 (IL-9) in human autoimmune diseases such as psoriasis and ulcerative colitis has been explored only very recently. However, their involvement in human rheumatoid arthritis (RA) is not conclusive. Pathogenesis of RA is complex and involves various T cell subsets and neutrophils. Here, we aimed at understanding the impact of IL-9 on infiltrating immune cells and their eventual role in synovial inflammation in RA.


In vitro stimulation of T cells was performed by engagement of anti-CD3 and anti-CD28 monoclonal antibodies. Flow cytometry was employed for measuring intracellular cytokine, RORγt in T cells, evaluating apoptosis of neutrophils. ELISA was used for measuring soluble cytokine, Western blot analysis and confocal microscopy were used for STAT3 phosphorylation and nuclear translocation.


We demonstrated synovial enrichment of Th9 cells and their positive correlation with disease activity (DAS28-ESR) in RA. Synovial IL-9 prolonged the survival of neutrophils, increased their matrix metalloprotienase-9 production and facilitated Th17 cell differentiation evidenced by induction of transcription factor RORγt and STAT3 phosphorylation. IL-9 also augmented the function of IFN-γ + and TNF-α + synovial T cells.


We provide evidences for critical role of IL-9 in disease pathogenesis and propose that targeting IL-9 may be an effective strategy to ameliorate synovial inflammation in RA. Inhibiting IL-9 may have wider impact on the production of pathogenic cytokines involved in autoimmune diseases including RA and may offer better control over the disease.
Additional file 1: Table S1. Demographic and clinical characteristics of RA patients (n = 28). (DOCX 12 kb)
Additional file 2: Figure S1. Identification and survival of synovial fluid neutrophils in presence of IL-9. Representative gating strategy for FACS plots are showing neutrophils from RA SF. Isolated neutrophils from RA SF were identified with positive staining for CD15 (dark area of FACS histogram, light-shaded area is isotype control, right upper panel FACS plot shows apoptosis of RA SF neutrophils at 0 hour). Apoptosis of neutrophils was measured with Annexin V staining in different culture conditions (media, SF, rIL-9 and anti-IL-9, FACS dot plots of lower panel). (TIF 360 kb)
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