Pathological effects of plasma from patients with thrombotic thrombocytopenic purpura on platelets and cultured vascular endothelial cells

Edward R. Burns, D. Zucker-Franklin

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

The pathological hallmarks of thrombotic thrombocytopenic purpura (TTP) include endothelial cell proliferation and subendothelial hyalin deposits in the microvasculature leading to symptomatic thrombotic occlusions. Plasma or sera from three consecutive patients with TTP were subjected to determine whether they induced endothelial injury and/or platelet activation, two pathogenic mechanisms that may account for this disorder. Sera were utilized in a microcytotoxicity assay against cultured human umbilical vein endothelial cells (EC). These cells were assessed ultrastructurally and with immunofluorescence techniques to ascertain the nature of inflicted cell damage. Control plasmas were obtained from healthy volunteers as well as patients with immune complex disease and the adult hemolytic uremic syndrome. In the presence of TTP serum, cell kill of 3H-proline-labeled EC averaged 42% versus 8.6% for control sera. Cytotoxicity induced by an IgG fraction of TTP sera averaged 70% versus 16.8% for control IgG. Removal of IgG by immune precipitation diminished cytotoxicity by 70%. Using indirect immunofluorescence IgG was detected in EC incubated with TTP serum but not on EC treated with control serum. Ultrastructural changes became apparent within 30m min after exposure of cultured EC to TTP serum. Virtually every cell developed numerous cytoplasmic inclusions rarely seen in EC in the presence of normal serum. Prolonged incubation with the TTP serum led to progressive cytolysis, terminating with complete cytoplasmic and nuclear degeneration. Plasma from all three patients with TTP caused spontaneous aggregation of normal washed platelets as monitored by aggregometry. No spontaneous aggregation occurred in response to control plasmas. These results indicate that the sera of the three TTP patients studied were able to mediate time-dependent immune destruction of human cultured endothelial cells and that their plasmas were capable of causing spontaneous aggregation of normal human platelets in vitro. It would seem likely that these mechanisms are also operative in vivo to produce the endothelial destruction as well as the thrombotic vascular occlusions seen in this disorder.

Original languageEnglish (US)
Pages (from-to)1030-1037
Number of pages8
JournalBlood
Volume60
Issue number4
StatePublished - 1982
Externally publishedYes

Fingerprint

Thrombotic Thrombocytopenic Purpura
Endothelial cells
Platelets
Blood Platelets
Endothelial Cells
Plasmas
Serum
Immunoglobulin G
Agglomeration
Cytotoxicity
Cultured Cells
Cell proliferation
Immune Complex Diseases
Antigen-Antibody Complex
Proline
Hemolytic-Uremic Syndrome
Hyalin
Assays
Inclusion Bodies
Human Umbilical Vein Endothelial Cells

ASJC Scopus subject areas

  • Immunology
  • Biochemistry
  • Hematology
  • Cell Biology

Cite this

Pathological effects of plasma from patients with thrombotic thrombocytopenic purpura on platelets and cultured vascular endothelial cells. / Burns, Edward R.; Zucker-Franklin, D.

In: Blood, Vol. 60, No. 4, 1982, p. 1030-1037.

Research output: Contribution to journalArticle

@article{2425c0e8641a4f11914da2b8faa73971,
title = "Pathological effects of plasma from patients with thrombotic thrombocytopenic purpura on platelets and cultured vascular endothelial cells",
abstract = "The pathological hallmarks of thrombotic thrombocytopenic purpura (TTP) include endothelial cell proliferation and subendothelial hyalin deposits in the microvasculature leading to symptomatic thrombotic occlusions. Plasma or sera from three consecutive patients with TTP were subjected to determine whether they induced endothelial injury and/or platelet activation, two pathogenic mechanisms that may account for this disorder. Sera were utilized in a microcytotoxicity assay against cultured human umbilical vein endothelial cells (EC). These cells were assessed ultrastructurally and with immunofluorescence techniques to ascertain the nature of inflicted cell damage. Control plasmas were obtained from healthy volunteers as well as patients with immune complex disease and the adult hemolytic uremic syndrome. In the presence of TTP serum, cell kill of 3H-proline-labeled EC averaged 42{\%} versus 8.6{\%} for control sera. Cytotoxicity induced by an IgG fraction of TTP sera averaged 70{\%} versus 16.8{\%} for control IgG. Removal of IgG by immune precipitation diminished cytotoxicity by 70{\%}. Using indirect immunofluorescence IgG was detected in EC incubated with TTP serum but not on EC treated with control serum. Ultrastructural changes became apparent within 30m min after exposure of cultured EC to TTP serum. Virtually every cell developed numerous cytoplasmic inclusions rarely seen in EC in the presence of normal serum. Prolonged incubation with the TTP serum led to progressive cytolysis, terminating with complete cytoplasmic and nuclear degeneration. Plasma from all three patients with TTP caused spontaneous aggregation of normal washed platelets as monitored by aggregometry. No spontaneous aggregation occurred in response to control plasmas. These results indicate that the sera of the three TTP patients studied were able to mediate time-dependent immune destruction of human cultured endothelial cells and that their plasmas were capable of causing spontaneous aggregation of normal human platelets in vitro. It would seem likely that these mechanisms are also operative in vivo to produce the endothelial destruction as well as the thrombotic vascular occlusions seen in this disorder.",
author = "Burns, {Edward R.} and D. Zucker-Franklin",
year = "1982",
language = "English (US)",
volume = "60",
pages = "1030--1037",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "4",

}

TY - JOUR

T1 - Pathological effects of plasma from patients with thrombotic thrombocytopenic purpura on platelets and cultured vascular endothelial cells

AU - Burns, Edward R.

AU - Zucker-Franklin, D.

PY - 1982

Y1 - 1982

N2 - The pathological hallmarks of thrombotic thrombocytopenic purpura (TTP) include endothelial cell proliferation and subendothelial hyalin deposits in the microvasculature leading to symptomatic thrombotic occlusions. Plasma or sera from three consecutive patients with TTP were subjected to determine whether they induced endothelial injury and/or platelet activation, two pathogenic mechanisms that may account for this disorder. Sera were utilized in a microcytotoxicity assay against cultured human umbilical vein endothelial cells (EC). These cells were assessed ultrastructurally and with immunofluorescence techniques to ascertain the nature of inflicted cell damage. Control plasmas were obtained from healthy volunteers as well as patients with immune complex disease and the adult hemolytic uremic syndrome. In the presence of TTP serum, cell kill of 3H-proline-labeled EC averaged 42% versus 8.6% for control sera. Cytotoxicity induced by an IgG fraction of TTP sera averaged 70% versus 16.8% for control IgG. Removal of IgG by immune precipitation diminished cytotoxicity by 70%. Using indirect immunofluorescence IgG was detected in EC incubated with TTP serum but not on EC treated with control serum. Ultrastructural changes became apparent within 30m min after exposure of cultured EC to TTP serum. Virtually every cell developed numerous cytoplasmic inclusions rarely seen in EC in the presence of normal serum. Prolonged incubation with the TTP serum led to progressive cytolysis, terminating with complete cytoplasmic and nuclear degeneration. Plasma from all three patients with TTP caused spontaneous aggregation of normal washed platelets as monitored by aggregometry. No spontaneous aggregation occurred in response to control plasmas. These results indicate that the sera of the three TTP patients studied were able to mediate time-dependent immune destruction of human cultured endothelial cells and that their plasmas were capable of causing spontaneous aggregation of normal human platelets in vitro. It would seem likely that these mechanisms are also operative in vivo to produce the endothelial destruction as well as the thrombotic vascular occlusions seen in this disorder.

AB - The pathological hallmarks of thrombotic thrombocytopenic purpura (TTP) include endothelial cell proliferation and subendothelial hyalin deposits in the microvasculature leading to symptomatic thrombotic occlusions. Plasma or sera from three consecutive patients with TTP were subjected to determine whether they induced endothelial injury and/or platelet activation, two pathogenic mechanisms that may account for this disorder. Sera were utilized in a microcytotoxicity assay against cultured human umbilical vein endothelial cells (EC). These cells were assessed ultrastructurally and with immunofluorescence techniques to ascertain the nature of inflicted cell damage. Control plasmas were obtained from healthy volunteers as well as patients with immune complex disease and the adult hemolytic uremic syndrome. In the presence of TTP serum, cell kill of 3H-proline-labeled EC averaged 42% versus 8.6% for control sera. Cytotoxicity induced by an IgG fraction of TTP sera averaged 70% versus 16.8% for control IgG. Removal of IgG by immune precipitation diminished cytotoxicity by 70%. Using indirect immunofluorescence IgG was detected in EC incubated with TTP serum but not on EC treated with control serum. Ultrastructural changes became apparent within 30m min after exposure of cultured EC to TTP serum. Virtually every cell developed numerous cytoplasmic inclusions rarely seen in EC in the presence of normal serum. Prolonged incubation with the TTP serum led to progressive cytolysis, terminating with complete cytoplasmic and nuclear degeneration. Plasma from all three patients with TTP caused spontaneous aggregation of normal washed platelets as monitored by aggregometry. No spontaneous aggregation occurred in response to control plasmas. These results indicate that the sera of the three TTP patients studied were able to mediate time-dependent immune destruction of human cultured endothelial cells and that their plasmas were capable of causing spontaneous aggregation of normal human platelets in vitro. It would seem likely that these mechanisms are also operative in vivo to produce the endothelial destruction as well as the thrombotic vascular occlusions seen in this disorder.

UR - http://www.scopus.com/inward/record.url?scp=0020000118&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020000118&partnerID=8YFLogxK

M3 - Article

C2 - 7052160

AN - SCOPUS:0020000118

VL - 60

SP - 1030

EP - 1037

JO - Blood

JF - Blood

SN - 0006-4971

IS - 4

ER -