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CpG oligonucleotide-mediated co-stimulation of mouse invariant natural killer T cells negatively regulates their activation status

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Abstract

Invariant natural killer T (iNKT) cells play important roles in antimicrobial defense and immune-regulation. We have previously shown that iNKT cells express certain toll-like receptors (TLR), and that TLR co-stimulation of iNKT cells in the presence of suboptimal concentrations of T cell receptor (TCR) agonists enhances cellular activation. In the present study, we investigated the regulatory effects of CpG oligonucleotides in mouse primary hepatic and splenic iNKT cells and in DN32.D3 iNKT cells. We show that CpG treatment of iNKT cells in the presence of higher concentrations of TCR agonists (α-GalCer or anti-CD3 mAb) results in the up-regulation of TLR9 in iNKT cells with a concurrent reduction in their cellular activation, as assessed by their production of IL-2, IL-4 and IFN-γ compared with controls. CpG-mediated down-regulation of iNKT cell activation has been found to depend, at least in part, on signaling by MyD88, a critical adapter moiety downstream of TLR9 signaling. Mechanistically, iNKT cells treated with CpG in the presence of TCR agonists show inhibition of MAPK signaling as determined by the levels of ERK1/2 and p38 MAPKs. Furthermore, CpG treatment leads to an increased induction of phosphatases, DUSP1 and SHP-1, that seem to impede MAPK and TCR signaling, resulting in the negative regulation of iNKT cell activation. Our findings therefore suggest a novel regulatory role for CpG in iNKT cells in the mediation of a negative feedback mechanism to control overactive iNKT cell responses and hence to avoid undesirable excessive immunopathology.

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Acknowledgements

The authors thank the NIH core tetramer facility at Emory University, Atlanta, Ga., USA for providing CD1d tetramers. They are also grateful to Emory University and University of Guelph Flow Cytometry Core Facility for help with flow cytometry. This work was partially supported by the National Institutes of Health (NIH) grants R01AI086133 and U19AI083019 to K.M.-K.

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  1. Alexander Ian Villanueva

    Present address: Sanofi Pasteur, North York, ON, Canada

  2. Jennifer T. Brisbin

    Present address: Ceva Animal Health, Guelph, ON, Canada

Authors and Affiliations

  1. Department of Pathobiology, University of Guelph, Guelph, ON, N1G2W1, Canada

    Raveendra R. Kulkarni, Alexander Ian Villanueva, Leah R. Read, Jennifer T. Brisbin & Shayan Sharif

  2. Department of Pediatrics and Emory Vaccine Center, Emory University School of Medicine, Atlanta, Ga., USA

    Siddhartha Kumar Bhaumik & Kaja Murali-Krishna

  3. ICGEB-Emory Vaccine Center, International Center for Genetic Engineering and Biotechnology, New Delhi, India

    Siddhartha Kumar Bhaumik & Kaja Murali-Krishna

  4. Department of Biomedical Sciences, University of Guelph, Guelph, ON, Canada

    Jonathan LaMarre

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  1. Raveendra R. Kulkarni
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The authors state that all applicable international, national, and/or institutional guidelines for the care and use of animals were followed, and all procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Kulkarni, R.R., Villanueva, A.I., Read, L.R. et al. CpG oligonucleotide-mediated co-stimulation of mouse invariant natural killer T cells negatively regulates their activation status. Cell Tissue Res 369, 541–554 (2017). https://doi.org/10.1007/s00441-017-2631-y

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