CXCL16-mediated cell recruitment to rheumatoid arthritis synovial tissue and murine lymph nodes is dependent upon the MAPK pathway

Authors

  • Jeffrey H. Ruth,

    1. University of Michigan Medical School, Ann Arbor, and Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Christian S. Haas,

    1. University of Michigan Medical School, Ann Arbor, and Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Christy C. Park,

    1. Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • M. Asif Amin,

    1. University of Michigan Medical School, Ann Arbor, and Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Rita J. Martinez,

    1. University of Michigan Medical School, Ann Arbor, and Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • G. Kenneth Haines III,

    1. Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Shiva Shahrara,

    1. Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Phillip L. Campbell,

    1. University of Michigan Medical School, Ann Arbor, and Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Alisa E. Koch

    Corresponding author
    1. University of Michigan Medical School, Ann Arbor, Northwestern University Feinberg School of Medicine, Chicago, Illinois, Veterans Administration Chicago Health Care Medical Center, Chicago, Illinois, and Ann Arbor Veterans Administration, Ann Arbor, Michigan
    • University of Michigan Medical School, Department of Medicine, Rheumatology Division, 1150 West Medical Center Drive, Ann Arbor, MI 48109-0680
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Abstract

Objective

Rheumatoid arthritis (RA) is characterized by profound mononuclear cell (MNC) recruitment into synovial tissue (ST), thought to be due in part to tumor necrosis factor α (TNFα), a therapeutic target for RA. Although chemokines may also be involved, the mechanisms remain unclear. We undertook this study to examine the participation of CXCL16, a novel chemokine, in recruitment of MNCs to RA ST in vivo and to determine the signal transduction pathways mediating this process.

Methods

Using a human RA ST–SCID mouse chimera, immunohistochemistry, enzyme-linked immunosorbent assay, real-time reverse transcription–polymerase chain reaction, flow cytometry, and in vitro chemotaxis assays, we defined the expression and function of CXCL16 and its receptor, CXCR6, as well as the signal transduction pathways utilized by them for MNC homing in vitro and in vivo.

Results

CXCL16 was markedly elevated in RA synovial fluid (SF) samples, being as high as 145 ng/ml. Intense macrophage and lining cell staining for CXCL16 in RA ST correlated with increased CXCL16 messenger RNA levels in RA ST compared with those in osteoarthritis and normal ST. By fluorescence-activated cell sorting analysis, one-half of RA SF monocytes and one-third of memory lymphocytes expressed CXCR6. In vivo recruitment of human MNCs to RA ST implanted in SCID mice occurred in response to intragraft injection of human CXCL16, a response similar to that induced by TNFα. Lipofection of MNCs with antisense oligodeoxynucleotides for ERK-1/2 resulted in a 50% decline in recruitment to engrafted RA ST and a 5-fold decline in recruitment to regional lymph nodes. Interestingly, RA ST fibroblasts did not produce CXCL16 in response to TNFα in vitro, suggesting that CXCL16 protein may function in large part independently of TNFα.

Conclusion

Taken together, these results point to a unique role for CXCL16 as a premier MNC recruiter in RA and suggest additional therapeutic possibilities, targeting CXCL16, its receptor, or its signaling pathways.

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