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Calcitriol attenuates TLR2/IL-33 signaling pathway to repress Th9 cell differentiation and potentially limits the pathophysiology of rheumatoid arthritis

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Abstract

There is limited information regarding the TLR2 signaling pathway involved in Th9 cell differentiation. The role of calcitriol in regulating TLR2-mediated Th9 cell development is unknown. Thus, we aimed to unravel the TLR2 signaling pathway in Th9 cells and its regulation by calcitriol. We have used n = 5–6 animals for each murine experiment. Human studies involved five healthy volunteers. Moreover, ten healthy individuals and ten RA patients were included in the study. Murine and human Th9 cells were treated with Calcitriol (100 nM) and Pam3CSK4 (2 µg/mL). The number of IL-9+ve cells was determined by flow cytometry. Real-time PCR was used to assess the gene expression. Serum 25(OH)D3 levels were determined by HPLC. We observed that TLR2 signals via IL-33/ST2 in Th9 cells. Increased TLR2 expression associated with increased IL9 expression and augmented disease severity in RA patients. Calcitriol attenuated TLR2 signaling in murine and human Th9 cells. Low serum vitamin D3 level negatively associated with increased IL-9 and TLR2 expression and disease severity in RA patients. Our data suggest a potential role of calcitriol to ameliorate the disease severity of RA patients.

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Abbreviations

TLRs:

Toll-like receptor

RA:

Rheumatoid arthritis

ESR:

Erythrocyte sedimentation rate

CRP:

C-reactive protein

RF:

Rheumatoid factor

25(OH)D3 :

25-Hydroxyvitamin D3; cholecalciferol

1,25(OH)2D3 :

1,25-Dihydroxyvitamin D3; calcitriol

Pam3CSK4 :

Pam3CysSerLys4

TCR:

T cell receptor

IL:

Interleukin

TGF-β:

Transforming growth factor-β

DAS28:

Disease Activity Score of 28 joints

Sfpi1:

Spleen focus forming virus proviral integration site 1

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Acknowledgements

SPV designed and performed the experiments, analyzed the data and wrote the manuscript. RG analyzed the data and wrote the manuscript. RNS provided the healthy and patients’ serum samples. RG also acknowledges Sudeepti Ratan Srivastava, King George’s Medical University, Lucknow for her assistance with the clinical samples. The flow cytometer and HPLC of the Central Research Facility, IIT Kharagpur is also acknowledged.

Funding

SPV acknowledges CSIR-UGC, Government of India Senior Research Fellowship. RG acknowledges DST-INNO-INDIGO (DST/IMRCD/EU/Inno Indigo/RA Detect/2015 (G)) and SERB (YSS/2015/001147), Government of India for financial support.

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Authors and Affiliations

  1. School of Bioscience, IIT Kharagpur, Kharagpur, 721302, West Bengal, India

    Shachi Pranjal Vyas & Ritobrata Goswami

  2. Department of Orthopedic Surgery, King George’s Medical University, Lucknow, Uttar Pradesh, 226003, India

    Rajeshwar Nath Srivastava

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  1. Shachi Pranjal Vyas
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Vyas, S.P., Srivastava, R.N. & Goswami, R. Calcitriol attenuates TLR2/IL-33 signaling pathway to repress Th9 cell differentiation and potentially limits the pathophysiology of rheumatoid arthritis. Mol Cell Biochem 476, 369–384 (2021). https://doi.org/10.1007/s11010-020-03914-4

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