In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of β2-glycoprotein I

Proof of concept

Yiannis Ioannou, Z. Romay-Penabad, C. Pericleous, I. Giles, E. Papalardo, Gracie Vargas, T. Shilagard, D. S. Latchman, D. A. Isenberg, A. Rahman, S. Pierangeli

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Objectives: In the antiphospholipid syndrome (APS), the immunodominant epitope for the majority of circulating pathogenic antiphospholipid antibodies (aPLs) is the N-terminal domain I (DI) of β2- glycoprotein I. We have previously shown that recombinant DI inhibits the binding of aPLs in fluid phase to immobilized native antigen, and that this inhibition is greater with the DI(D8S/D9G) mutant and absent with the DI(R39S) mutant. Hence, we hypothesized that DI and DI(D8S/D9G) would inhibit aPL-induced pathogenicity in vivo. Methods: C57BL/6 mice (n = 5, each group) were injected with purified IgG derived from APS patients (IgG-APS, 500 μg) or IgG from normal healthy serum (IgG-NHS) and either recombinant DI, DI(R39S), DI(D8S/ D9G), or an irrelevant control peptide (at 10-40 μg). Outcome variables measured were femoral vein thrombus dynamics in treated and control groups following standardized vessel injury, expression of vascular cell adhesion molecule-1 (VCAM-1) on the aortic endothelial surface, and tissue factor (TF) activity in murine macrophages. Results: IgG-APS significantly increased thrombus size as compared with IgG-NHS. The IgG-APS thrombus enhancement effect was abolished in mice pretreated with recombinant DI (P ≤ 0.0001) and DI(D8S/D9G) (P ≤ 0.0001), but not in those treated with DI(R39S) or control peptide. This inhibitory effect by DI was dose-dependent, and at lower doses DI(D8S/ D9G) was a more potent inhibitor of thrombosis than wild-type DI (P ≤ 0.01). DI also inhibited IgG-APS induction of VCAM-1 on the aortic endothelial surface and TF production by murine macrophages. Conclusion: Our findings in this proof-of-concept study support the development of recombinant DI or the novel variant DI(D8S/D9G) as a potential future therapeutic agent for APS.

Original languageEnglish (US)
Pages (from-to)833-842
Number of pages10
JournalJournal of Thrombosis and Haemostasis
Volume7
Issue number5
DOIs
StatePublished - 2009

Fingerprint

Antiphospholipid Antibodies
Antiphospholipid Syndrome
Virulence
Glycoproteins
Immunoglobulin G
Thrombosis
Vascular Cell Adhesion Molecule-1
Thromboplastin
Endothelium
Macrophages
Immunodominant Epitopes
Peptides
Femoral Vein
Protein Domains
Serum
Inbred C57BL Mouse
Antigens
Control Groups
Wounds and Injuries

Keywords

  • β-glycoprotein I
  • Antiphospholipid antibodies
  • Antiphospholipid syndrome
  • thrombosis
  • Tissue factor
  • Vascular cell adhesion molecule-1

ASJC Scopus subject areas

  • Hematology

Cite this

In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of β2-glycoprotein I : Proof of concept. / Ioannou, Yiannis; Romay-Penabad, Z.; Pericleous, C.; Giles, I.; Papalardo, E.; Vargas, Gracie; Shilagard, T.; Latchman, D. S.; Isenberg, D. A.; Rahman, A.; Pierangeli, S.

In: Journal of Thrombosis and Haemostasis, Vol. 7, No. 5, 2009, p. 833-842.

Research output: Contribution to journalArticle

Ioannou, Y, Romay-Penabad, Z, Pericleous, C, Giles, I, Papalardo, E, Vargas, G, Shilagard, T, Latchman, DS, Isenberg, DA, Rahman, A & Pierangeli, S 2009, 'In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of β2-glycoprotein I: Proof of concept', Journal of Thrombosis and Haemostasis, vol. 7, no. 5, pp. 833-842. https://doi.org/10.1111/j.1538-7836.2009.03316.x
Ioannou, Yiannis ; Romay-Penabad, Z. ; Pericleous, C. ; Giles, I. ; Papalardo, E. ; Vargas, Gracie ; Shilagard, T. ; Latchman, D. S. ; Isenberg, D. A. ; Rahman, A. ; Pierangeli, S. / In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of β2-glycoprotein I : Proof of concept. In: Journal of Thrombosis and Haemostasis. 2009 ; Vol. 7, No. 5. pp. 833-842.
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T1 - In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of β2-glycoprotein I

T2 - Proof of concept

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AU - Romay-Penabad, Z.

AU - Pericleous, C.

AU - Giles, I.

AU - Papalardo, E.

AU - Vargas, Gracie

AU - Shilagard, T.

AU - Latchman, D. S.

AU - Isenberg, D. A.

AU - Rahman, A.

AU - Pierangeli, S.

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AB - Objectives: In the antiphospholipid syndrome (APS), the immunodominant epitope for the majority of circulating pathogenic antiphospholipid antibodies (aPLs) is the N-terminal domain I (DI) of β2- glycoprotein I. We have previously shown that recombinant DI inhibits the binding of aPLs in fluid phase to immobilized native antigen, and that this inhibition is greater with the DI(D8S/D9G) mutant and absent with the DI(R39S) mutant. Hence, we hypothesized that DI and DI(D8S/D9G) would inhibit aPL-induced pathogenicity in vivo. Methods: C57BL/6 mice (n = 5, each group) were injected with purified IgG derived from APS patients (IgG-APS, 500 μg) or IgG from normal healthy serum (IgG-NHS) and either recombinant DI, DI(R39S), DI(D8S/ D9G), or an irrelevant control peptide (at 10-40 μg). Outcome variables measured were femoral vein thrombus dynamics in treated and control groups following standardized vessel injury, expression of vascular cell adhesion molecule-1 (VCAM-1) on the aortic endothelial surface, and tissue factor (TF) activity in murine macrophages. Results: IgG-APS significantly increased thrombus size as compared with IgG-NHS. The IgG-APS thrombus enhancement effect was abolished in mice pretreated with recombinant DI (P ≤ 0.0001) and DI(D8S/D9G) (P ≤ 0.0001), but not in those treated with DI(R39S) or control peptide. This inhibitory effect by DI was dose-dependent, and at lower doses DI(D8S/ D9G) was a more potent inhibitor of thrombosis than wild-type DI (P ≤ 0.01). DI also inhibited IgG-APS induction of VCAM-1 on the aortic endothelial surface and TF production by murine macrophages. Conclusion: Our findings in this proof-of-concept study support the development of recombinant DI or the novel variant DI(D8S/D9G) as a potential future therapeutic agent for APS.

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