Article

The Act1 D10N missense variant impairs CD40 signaling in human B-cells

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Abstract

The TRAF3IP2 gene resides within one of at least 63 psoriasis susceptibility loci and encodes Act1, an adapter protein involved in IL-17 receptor and CD40 signaling pathways. TRAF3IP2 is distinctive (among <10% of candidate susceptibility genes) in that a strongly disease-associated variant encodes a missense SNP predicted to be functionally relevant (SNP rs33980500 C/T encoding Act1 pD10N). As assessed by flow cytometry, Act1 protein was expressed at the highest levels in monocytes, with lower levels in T-cells and B-cells. However, monocytes, T-cells and B-cells failed to respond to IL-17A stimulation of PBMC, as measured by flow cytometric determination of NF-κB phospho-p65. As an alternative stimulus, we treated PBMCs with trimerized recombinant human CD40L and assessed p65, p38 and Erk phosphorylation in CD19+ B-cells as a function of D10N genotype. The increase of phosphorylated p65, p38, and Erk was well-correlated across individuals, and CD40L-induced phosphorylation of p65, p38, and Erk was significantly attenuated in B-cells from Act1 D10N homozygotes, compared to heterozygotes and nullizygotes. Our results indicate that the Act1 D10N variant is a relevant genetic determinant of CD40L responsiveness in human B-cells, with the risk allele being associated with lower B-cell responses in an acute signaling context.

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Acknowledgements

We thank the volunteers who provided blood samples for this study, and Dr. Andrew Johnston for advice on flow cytometry. This research was supported by National Institutes of Health (NIH) R01 grants AR042742, AR050511, AR054966, AR062382, and AR065183 to JTE. We also acknowledge generous support from the Dawn and Dudley Holmes Memorial Fund and the Babcock Endowment Fund to the Department of Dermatology at the University of Michigan. JTE is supported by the Ann Arbor Veterans Affairs Hospital.

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Author notes

  1. Ning Yu and Sylviane Lambert contributed equally to this work

Affiliations

  1. Department of Dermatology, University of Michigan, Ann Arbor, MI, USA

    • Ning Yu
    • , Sylviane Lambert
    • , Joshua Bornstein
    • , Rajan P. Nair
    • , Charlotta Enerbäck
    •  & James T. Elder
  2. Department of Dermatology, Shanghai Skin Disease Hospital, Shanghai, China

    • Ning Yu
  3. Department of Dermatology, Linköping University, Linköping, Sweden

    • Charlotta Enerbäck
  4. Ann Arbor Veterans Affairs Health System, Ann Arbor, MI, USA

    • James T. Elder

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Conflict of interest

The authors declare that they have no competing interests.

Corresponding authors

Correspondence to Ning Yu or James T. Elder.