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Membrane-Bound Toll-Like Receptors are Overexpressed in Peripheral Blood and Synovial Fluid Mononuclear Cells of Enthesitis-Related Arthritis Category of Juvenile Idiopathic Arthritis (JIA–ERA) Patients and Lead to Secretion of Inflammatory Mediators

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Abstract

We examined expression and function of TLRs in enthesitis-related arthritis (ERA) patients. RNA levels of TLR1, TLR3, and TLRs 5–8 were measured in 24 ERA peripheral blood mononuclear cells (PBMC), 18 synovial fluid mononuclear cells (SFMC), and IRAK1, IRAK4, TRIF, TRAF3, and TRAF6 in 18 PBMC and 10 SFMC. IL-6 and IL-8 were measured in supernatants from ERA PBMC (n = 7), SFMC (n = 3), and healthy PBMC (n = 5) cultured with ligands for TLR1/2 (Pam 3-cys), TLR3 (poly I:C), TLR5 (flagellin), and TLR2/6 (zymosan). TLRs 1, 3, 5, and 6 were measured in whole blood (n = 20 ERA, seven healthy) and SFMC (n = 2) by flow cytometry. ERA PBMC compared to healthy PBMC and SFMC compared to ERA PBMC had higher RNA expression of TLR1, TLR3, TLR5, TLR6, IRAK1, IRAK4, TRIF, TRAF3, and TRAF6. TLR7 and TLR8 RNA expression was similar in all study groups. IL-6 and IL-8 levels were higher in stimulated ERA SFMC compared to ERA PBMC and in ERA PBMC compared to control PBMC. TLRs 1, 3, and 6 were also overexpressed at the protein level.

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Acknowledgments

This study was supported by a grant from the Indian Council of Medical Research and Department of Biotechnology, Government of India to A.A. A.M. is supported by Council of Scientific and Industrial Research–Senior Research Fellowship.

Conflict of Interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India

    Arpita Myles, Mujeeb T. Rahman & Amita Aggarwal

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  1. Arpita Myles
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  2. Mujeeb T. Rahman
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  3. Amita Aggarwal
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Corresponding author

Correspondence to Amita Aggarwal.

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Supplementary Fig. S1

RNA levels of TLRs normalized to 18S rRNA. a Fold change in patient PBMC (n = 24) relative to healthy PBMC. b Fold change in patient (n = 7) SFMC relative to corresponding PBMC (JPEG 58 kb)

Supplementary Fig. S2

RNA levels of adaptors normalized to GAPDH. a Fold change in patient PBMC (n = 18) relative to healthy PBMC. b Fold change in patient (n = 10) SFMC relative to corresponding PBMC (JPEG 60 kb)

Supplementary Fig. S3

Flow cytometry histograms comparing expression of various toll-like receptors in whole blood upon blocking Fc receptors with 10% human serum. Samples were treated with either HI Serum heat inactivated serum, UT Serum untreated serum (used without activation), or left untreated (no serum) (JPEG 214 kb)

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Myles, A., Rahman, M.T. & Aggarwal, A. Membrane-Bound Toll-Like Receptors are Overexpressed in Peripheral Blood and Synovial Fluid Mononuclear Cells of Enthesitis-Related Arthritis Category of Juvenile Idiopathic Arthritis (JIA–ERA) Patients and Lead to Secretion of Inflammatory Mediators. J Clin Immunol 32, 488–496 (2012). https://doi.org/10.1007/s10875-011-9640-5

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  • DOI: https://doi.org/10.1007/s10875-011-9640-5

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