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Kidney Week

Abstract: FR-PO860

MHC Class II Ligation on Glomerular Endothelial Cells Upregulates Adhesion Molecules and Chemoattractants

Session Information

  • Transplantation: Basic
    October 26, 2018 | Location: Exhibit Hall, San Diego Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Transplantation

  • 1801 Transplantation: Basic

Authors

  • Panzer, Sarah E., University of Wisconsin Madison, Madison, Wisconsin, United States
  • Wilson, Nancy A., University of Wisconsin Madison, Madison, Wisconsin, United States
  • Dylewski, James F., University of Colorado Hospital, Aurora, Colorado, United States
  • Djamali, Arjang, University of Wisconsin Madison, Madison, Wisconsin, United States
  • Blaine, Judith, University of Colorado Denver , Aurora, Colorado, United States
Background

Microvascular injury and leukocyte margination across endothelial cells are key pathologic processes in kidney allograft rejection. The mechanisms of glomerular endothelial cell injury and leukocyte infiltration remain unclear. We hypothesize MHC class II ligation on glomerular endothelial cells will result in upregulation of adhesion molecules and chemoattractants.

Methods

A Bl/6 mouse glomerular endothelial cell (MGEC) line was stimulated with IFNγ to upregulate MHC class I and II; then incubated with anti-MHC class II antibodies or plasma from balb/c mice sensitized with Bl/6 splenocytes. Expression of adhesion molecules were assessed by flow cytometry. Gene expression was measured by RT2 profiler and Taqman RT PCR. Cytokine expression was analyzed in culture supernatants with LegendPlex.

Results

MGEC stimulated with IFNγ upregulated gene expression of TRAIL, RANTES, MCP-1, caspase1, beta2microglobulin, Vcam1, IL-6 and Icam1 (A). IFNγ stimulation of MGEC increased surface expression of MHC class I, MHC class II, VCAM1, and ICAM1; but not P-selectin (B). Subsequent ligation with MHC class II antibody transiently increased surface expression of P-selectin at 6 hours; while sensitized plasma resulted in a greater increase in P-selectin, which persisted at 24 hours (C). MGEC secretion of chemoattractants (MCP-1 and RANTES) increased with IFNγ stimulation and was further enhanced with MHC II ligation (D). Adhesion molecules and MHC co-localize on the cell surface (E).

Conclusion

MHC II crosslinking on MGEC resulted in an upregulation of P-selectin as well as chemoattractants. MHC molecules and adhesion molecules are co-localized on the endothelial cell surface. Future studies will examine therapeutic targets, such as inhibitors of mTOR (inhibition of ICAM1 clustering) and Nox4 (TRAIL pathway).

Funding

  • Other NIH Support