A need for standardization of the anti-endothelial-cell antibody test

Youinou, Pierre; Meroni, Pier Luigi; Khamashta, Munther A.; Shoenfeld, Yehuda

doi:10.1016/0167-5699(95)80001-8

Cited by 57 publications

(38 citation statements)

References 7 publications

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“…However, these methods favor the detection of cytosol proteins (16). Flow cytometry, a method which detects antigens on the cell surface, is more informative for examining the role of anti-EC Abs in chronic rejection (13).…”

Section: Introductionmentioning

confidence: 99%

Non-HLA-Type Endothelial Cell Reactive Alloantibodies in Pre-Transplant Sera of Kidney Recipients Trigger Apoptosis

Bas‐Bernardet

Hourmant

Coupel

et al. 2003

American Journal of Transplantation

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The vascular endothelium of transplanted organs represents an important target for allograft-directed immune responses. Although HLA antigens expressed on graft endothelial cells (EC) can become targets of the host immune response, the role of other, non-HLA-encoded EC antigens has been proposed but is still unclear. The aim of this study was to investigate the presence of and to characterize anti-EC antibodies (AECA) in 57 kidney transplant recipients according to their HLAimmunization status. Flow cytometry in pretransplant sera was used to detect AECA reactive with surface antigens on ABO and HLA-typed primary cultures of arterial ECs, stimulated or not with tumor necrosis factor-a (TNFa) or interferon-g (IFNg). FACS analysis revealed the presence of AECA in 47% of HLA-sensitized (PRA = 10%: mostly IgG) vs. 16.0% in nonsensitized patients (PRA < 10%) (p < 0.02). No significant correlation was found between the presence of AECA and acute rejection occurrence and graft outcome. Non-HLA reactive AECA are directed against TNFa-and IFNg-inducible membrane molecule(s), and react with two predominant antigens of~35 kDa and~50 kDa expressed on ECs but not on B cells. Binding of AECA decreases in vitro EC viability by 50-60% by promoting EC apoptosis, as demonstrated by DNA fragmentation assays.

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Section: Introductionmentioning

confidence: 99%

Non-HLA-Type Endothelial Cell Reactive Alloantibodies in Pre-Transplant Sera of Kidney Recipients Trigger Apoptosis

Bas‐Bernardet

Hourmant

Coupel

et al. 2003

American Journal of Transplantation

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“…These autoantibodies (Ab) have since been reported in a variety of clinical settings associated with vasculitis (2), such as systemic sclerosis (SS). Although the results obtained in a given disease vary from one study to another (3), the AECA test shows promise as a sensitive indicator of endothelial cell (EC) injury (4). In fact, AECAs represent an extremely heterogeneous family of Ab reacting with different structures on EC (5), the effects of which remain to be defined.…”

Section: Introductionmentioning

confidence: 99%

The binding of some human antiendothelial cell antibodies induces endothelial cell apoptosis.

Bordron¹,

Dueymes²,

Levy³

et al. 1998

J. Clin. Invest.

202

130

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The pathogenic role of antiendothelial cell antibodies (AECA) remains unclear. They are frequently associated with antibodies to anionic phospholipids (PL), such as phosphatidylserine (PS), which is difficult to reconcile with the distribution of PL molecular species within the plasma membrane. Since it is already known that PS is transferred to the outer face of the membrane as a preclude to apoptosis, the possibility exists that apoptosis is initiated by AECA. AECA-positive/anti-PL antibody-negative sera from eight patients with systemic sclerosis (SS) and 21 control patients were evaluated. Endothelial cells (EC) were incubated with AECA and the exposure of PS was established through the binding of annexin V. Hypoploid cell enumeration, DNA fragmentation, and optical and ultrastructural analyses of EC were used to confirm apoptosis. Incubation of EC with AECA derived from six of eight patients with SS led to the expression of PS on the surface of the cells. This phenomenon was significantly more frequent in SS ( P Ͻ 0.04) than in control diseases. The redistribution of plasma membrane PS preceded other events associated with apoptosis: hypoploidy, DNA fragmentation, and morphology characteristic for apoptosis. Apoptosis-inducing AECA did not recognize the Fas receptor. We conclude that AECA may be pathogenic by inducing apoptosis. (

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“…line normalized for protein and DNA contents could allow better interlaboratory agreement in the measurement of AECA titers by ELISA and allow the data obtained for the same sera processed by different assays to be compared, as recommended by Youinou et al (24). In addition, this could contribute to a reduction in the important discrepancies in associations between AECA titers and clinical conditions ascribed to antigen variability (13,2,9).…”

Section: Discussionmentioning

confidence: 99%

“…Spurious increases in AECA titers may occur, e.g., due to anti-DNA autoantibodies, depending on an important cross-reactivity against endothelial cells (4); anti-heparan sulfate antibodies (16); or heterophile antibodies to the bovine serum proteins involved in the assay (18). One of the main problems in this field is the lack of agreement on a standardized method for detection of AECAs, with subsequent difficulty with interlaboratory comparisons (13,24). In addition, in rheumatoid arthritis and Felty's syndrome, rheumatoid factor (RF) has been shown to increase nonspecific immunoglobulin binding to endothelial cells, with subsequent pitfalls in assays with RF-containing sera developed with endothelial cells (15).…”

mentioning

confidence: 99%

Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors

Drouet

Nissou²,

Ponard³

et al. 2003

Clin Vaccine Immunol

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A need for standardization of the anti-endothelial-cell antibody test

Cited by 57 publications

References 7 publications

Non-HLA-Type Endothelial Cell Reactive Alloantibodies in Pre-Transplant Sera of Kidney Recipients Trigger Apoptosis

Non-HLA-Type Endothelial Cell Reactive Alloantibodies in Pre-Transplant Sera of Kidney Recipients Trigger Apoptosis

The binding of some human antiendothelial cell antibodies induces endothelial cell apoptosis.

Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors

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