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Antibodies Against Endothelial Cells

Antibodies Against Endothelial Cells. Peter Stastny Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas. C4d staining in peritubular capilaries in acute humoral rejection. Preparation of Umbilical Vein Endothelial Cells.

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Antibodies Against Endothelial Cells

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  1. Antibodies Against Endothelial Cells Peter Stastny Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas

  2. C4d staining in peritubular capilaries in acute humoral rejection

  3. Preparation of Umbilical Vein Endothelial Cells

  4. Antibodies to Endothelial Cells (E) and Matching Cord Blood Lymphocytes (L) From Moraes and Stastny, Histocompatibility Testing 1975.

  5. Flow Cytometry with Endothelial Cells • Two patterns of reactions with a panel: • All positive - Probable auto-antibody • Some positive and others negative – • Probable allo-antibody

  6. Serum DF Reactive with Vimentin: All Positive

  7. Serum AJ Reactive with KRT1: Some Positive and some Negative

  8. Immunoprecipitation and Protein Identification Endothelial surfaceaffinity-purified patient IgG Reactive endothelialcell lysate Immunoprecipitate Eluate Isolate bands from SDS-PAGE Mass spectrometry Protein identification

  9. Antibodies found in Patient Sera after Transplant Rejection Auto-Antibodies Pt. Size Alpha-tubulin SW 53 kD Nucleolin WW 100 kD Vimentin DF 55 kD Allo-Antibodies KRT-1 AJ 67 kD MICA MD 53 kD Possible Allo-Antibodies EIF2A DM 67 kD ErbB3 protein 1 EH 44 kD ERLIN2 WW 35 kD

  10. Surface Expression on HUVEC Nucleolin Vimentin Keratin-1 MICA

  11. Frequency of MORA Alleles in Four Populations

  12. Summary • Our first paper about endothelial cell antibodies was published in 1975, with Roberto Moraes. • Cytotoxic endothelial-specific antibodies, defined as E+L- reactions, were found in over half of recipients after rejection. • Flow-cytometry showed monomorphic and polymorphic patterns of panel cell staining. • Immuno-precipitation and protein identification was used to determine target antigens on the surface of endothelial cells. • MORA (KRT1) antigens are polymorphic and antibodies can be determined by ELISA. • Thank you!

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