Antibody to human leukocyte antigen triggers endothelial exocytosis

Munekazu Yamakuchi, Nancy C. Kirkiles-Smith, Marcella Ferlito, Scott J. Cameron, Clare Bao, Karen Fox-Talbot, Barbara A. Wasowska, William M. Baldwin, Jordan S. Pober, Charles J. Lowenstein

Research output: Contribution to journalArticle

Abstract

Although antibodies to HLA play a role in the pathogenesis of diseases processes such as rejection of transplanted organs, the precise mechanisms by which antibodies cause tissue injury are not completely understood. We hypothesized that antibodies to host tissues cause inflammation in part by activating endothelial exocytosis of granules that contain prothrombotic mediators such as von Willebrand Factor (VWF) and proinflammatory mediators such as P-selectin. To test this hypothesis, we treated human endothelial cells with murine monoclonal antibody W6/32 to HLA class I and then measured exocytosis by the release of VWF and the externalization of P-selectin. Antibody to MLA activates endothelial exocytosis in a dose-dependent manner over time. The biologically active complement split product, C5a, adds a slight but significant increase to antibody induction of exocytosis. Antibody to HLA alone or with C5a did not damage the cells. Cross-linking of HLA appears to play a role in the ability of antibody to activate exocytosis, because the W6/32 monovalent Fab fragment did not activate VWF release, but the bivalent F(ab′)2 was effective in triggering exocytosis. To explore the in vivo effects of antibody upon graft injury, we infused W6/32 F(ab′)2 antibody to human HLA into severe combined immunodeficient/beige mice that had been transplanted with human skin grafts. Antibody to HLA activated exocytosis and inflammation in human skin grafts. Our data show that antibody to host antigens can activate human endothelial cell exocytosis and leukocyte trafficking. By triggering vascular inflammation, antibody activation of exocytosis may play a role in transplant rejection.

Original languageEnglish (US)
Pages (from-to)1301-1306
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number4
DOIs
StatePublished - Jan 23 2007

Fingerprint

Exocytosis
HLA Antigens
Antibodies
von Willebrand Factor
P-Selectin
Inflammation
Transplants
Endothelial Cells
Skin
Immunoglobulin Fab Fragments
SCID Mice
Wounds and Injuries
Graft Rejection
Blood Vessels
Leukocytes
Monoclonal Antibodies

Keywords

  • Nitric oxide
  • Rejection
  • Transplant
  • Vasculopathy
  • Weibel-Palade body

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Yamakuchi, M., Kirkiles-Smith, N. C., Ferlito, M., Cameron, S. J., Bao, C., Fox-Talbot, K., ... Lowenstein, C. J. (2007). Antibody to human leukocyte antigen triggers endothelial exocytosis. Proceedings of the National Academy of Sciences of the United States of America, 104(4), 1301-1306. https://doi.org/10.1073/pnas.0602035104

Antibody to human leukocyte antigen triggers endothelial exocytosis. / Yamakuchi, Munekazu; Kirkiles-Smith, Nancy C.; Ferlito, Marcella; Cameron, Scott J.; Bao, Clare; Fox-Talbot, Karen; Wasowska, Barbara A.; Baldwin, William M.; Pober, Jordan S.; Lowenstein, Charles J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 4, 23.01.2007, p. 1301-1306.

Research output: Contribution to journalArticle

Yamakuchi, M, Kirkiles-Smith, NC, Ferlito, M, Cameron, SJ, Bao, C, Fox-Talbot, K, Wasowska, BA, Baldwin, WM, Pober, JS & Lowenstein, CJ 2007, 'Antibody to human leukocyte antigen triggers endothelial exocytosis', Proceedings of the National Academy of Sciences of the United States of America, vol. 104, no. 4, pp. 1301-1306. https://doi.org/10.1073/pnas.0602035104
Yamakuchi, Munekazu ; Kirkiles-Smith, Nancy C. ; Ferlito, Marcella ; Cameron, Scott J. ; Bao, Clare ; Fox-Talbot, Karen ; Wasowska, Barbara A. ; Baldwin, William M. ; Pober, Jordan S. ; Lowenstein, Charles J. / Antibody to human leukocyte antigen triggers endothelial exocytosis. In: Proceedings of the National Academy of Sciences of the United States of America. 2007 ; Vol. 104, No. 4. pp. 1301-1306.
@article{03c8ea727259468b96e9cfcec2d47bb5,
title = "Antibody to human leukocyte antigen triggers endothelial exocytosis",
abstract = "Although antibodies to HLA play a role in the pathogenesis of diseases processes such as rejection of transplanted organs, the precise mechanisms by which antibodies cause tissue injury are not completely understood. We hypothesized that antibodies to host tissues cause inflammation in part by activating endothelial exocytosis of granules that contain prothrombotic mediators such as von Willebrand Factor (VWF) and proinflammatory mediators such as P-selectin. To test this hypothesis, we treated human endothelial cells with murine monoclonal antibody W6/32 to HLA class I and then measured exocytosis by the release of VWF and the externalization of P-selectin. Antibody to MLA activates endothelial exocytosis in a dose-dependent manner over time. The biologically active complement split product, C5a, adds a slight but significant increase to antibody induction of exocytosis. Antibody to HLA alone or with C5a did not damage the cells. Cross-linking of HLA appears to play a role in the ability of antibody to activate exocytosis, because the W6/32 monovalent Fab fragment did not activate VWF release, but the bivalent F(ab′)2 was effective in triggering exocytosis. To explore the in vivo effects of antibody upon graft injury, we infused W6/32 F(ab′)2 antibody to human HLA into severe combined immunodeficient/beige mice that had been transplanted with human skin grafts. Antibody to HLA activated exocytosis and inflammation in human skin grafts. Our data show that antibody to host antigens can activate human endothelial cell exocytosis and leukocyte trafficking. By triggering vascular inflammation, antibody activation of exocytosis may play a role in transplant rejection.",
keywords = "Nitric oxide, Rejection, Transplant, Vasculopathy, Weibel-Palade body",
author = "Munekazu Yamakuchi and Kirkiles-Smith, {Nancy C.} and Marcella Ferlito and Cameron, {Scott J.} and Clare Bao and Karen Fox-Talbot and Wasowska, {Barbara A.} and Baldwin, {William M.} and Pober, {Jordan S.} and Lowenstein, {Charles J.}",
year = "2007",
month = "1",
day = "23",
doi = "10.1073/pnas.0602035104",
language = "English (US)",
volume = "104",
pages = "1301--1306",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "4",

}

TY - JOUR

T1 - Antibody to human leukocyte antigen triggers endothelial exocytosis

AU - Yamakuchi, Munekazu

AU - Kirkiles-Smith, Nancy C.

AU - Ferlito, Marcella

AU - Cameron, Scott J.

AU - Bao, Clare

AU - Fox-Talbot, Karen

AU - Wasowska, Barbara A.

AU - Baldwin, William M.

AU - Pober, Jordan S.

AU - Lowenstein, Charles J.

PY - 2007/1/23

Y1 - 2007/1/23

N2 - Although antibodies to HLA play a role in the pathogenesis of diseases processes such as rejection of transplanted organs, the precise mechanisms by which antibodies cause tissue injury are not completely understood. We hypothesized that antibodies to host tissues cause inflammation in part by activating endothelial exocytosis of granules that contain prothrombotic mediators such as von Willebrand Factor (VWF) and proinflammatory mediators such as P-selectin. To test this hypothesis, we treated human endothelial cells with murine monoclonal antibody W6/32 to HLA class I and then measured exocytosis by the release of VWF and the externalization of P-selectin. Antibody to MLA activates endothelial exocytosis in a dose-dependent manner over time. The biologically active complement split product, C5a, adds a slight but significant increase to antibody induction of exocytosis. Antibody to HLA alone or with C5a did not damage the cells. Cross-linking of HLA appears to play a role in the ability of antibody to activate exocytosis, because the W6/32 monovalent Fab fragment did not activate VWF release, but the bivalent F(ab′)2 was effective in triggering exocytosis. To explore the in vivo effects of antibody upon graft injury, we infused W6/32 F(ab′)2 antibody to human HLA into severe combined immunodeficient/beige mice that had been transplanted with human skin grafts. Antibody to HLA activated exocytosis and inflammation in human skin grafts. Our data show that antibody to host antigens can activate human endothelial cell exocytosis and leukocyte trafficking. By triggering vascular inflammation, antibody activation of exocytosis may play a role in transplant rejection.

AB - Although antibodies to HLA play a role in the pathogenesis of diseases processes such as rejection of transplanted organs, the precise mechanisms by which antibodies cause tissue injury are not completely understood. We hypothesized that antibodies to host tissues cause inflammation in part by activating endothelial exocytosis of granules that contain prothrombotic mediators such as von Willebrand Factor (VWF) and proinflammatory mediators such as P-selectin. To test this hypothesis, we treated human endothelial cells with murine monoclonal antibody W6/32 to HLA class I and then measured exocytosis by the release of VWF and the externalization of P-selectin. Antibody to MLA activates endothelial exocytosis in a dose-dependent manner over time. The biologically active complement split product, C5a, adds a slight but significant increase to antibody induction of exocytosis. Antibody to HLA alone or with C5a did not damage the cells. Cross-linking of HLA appears to play a role in the ability of antibody to activate exocytosis, because the W6/32 monovalent Fab fragment did not activate VWF release, but the bivalent F(ab′)2 was effective in triggering exocytosis. To explore the in vivo effects of antibody upon graft injury, we infused W6/32 F(ab′)2 antibody to human HLA into severe combined immunodeficient/beige mice that had been transplanted with human skin grafts. Antibody to HLA activated exocytosis and inflammation in human skin grafts. Our data show that antibody to host antigens can activate human endothelial cell exocytosis and leukocyte trafficking. By triggering vascular inflammation, antibody activation of exocytosis may play a role in transplant rejection.

KW - Nitric oxide

KW - Rejection

KW - Transplant

KW - Vasculopathy

KW - Weibel-Palade body

UR - http://www.scopus.com/inward/record.url?scp=33846586525&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846586525&partnerID=8YFLogxK

U2 - 10.1073/pnas.0602035104

DO - 10.1073/pnas.0602035104

M3 - Article

C2 - 17229850

AN - SCOPUS:33846586525

VL - 104

SP - 1301

EP - 1306

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 4

ER -