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1. Anti-phospholipid IgG antibodies detected by line immunoassay differentiate patients with anti-phospholipid syndrome and other autoimmune diseases.

Author

Nalli, Cecilia, Somma, Valentina, Andreoli, Laura, Büttner, Thomas, Schierack, Peter, Mahler, Michael, Roggenbuck, Dirk, and Tincani, Angela

Subjects
*IMMUNOGLOBULIN G, *AUTOIMMUNE disease treatment, *ANTIPHOSPHOLIPID syndrome treatment, *GLYCOPROTEINS, *PHOSPHATIDYLGLYCEROL, *IMMUNOASSAY
Abstract

Purpose: Anti-phospholipid antibodies (aPL) analyzed by line immunoassay (LIA) can recognize beta2-glycoprotein I (β2GPI) domain 1 (D1) epitopes depending on β2GPI binding to distinct phospholipids. The aPL LIA was compared with consensus ELISA to investigate whether both techniques can discriminate anti-phospholipid syndrome (APS) patients from aPL-positive, systemic autoimmune rheumatic diseases (SARD) patients without clinical symptoms of APS and controls.Methods: Thirty-four APS patients (14 arterial/venous thrombosis, 16 pregnancy morbidity, and 4 both), 41 patients with SARD lacking clinical APS criteria but demonstrating positivity for anti-β2GPI (aβ2GPI) IgG, and 20 healthy subjects (HS) were tested for aPL to cardiolipin (aCL), phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol (aPG), phosphatidylinositol, phosphatidylserine, β2GPI, prothrombin, and annexin V by LIA. Samples were also tested for aCL, aβ2GPI, aβ2GPI-domain 1 (aD1), and aβ2GPI-domains 4-5 (aD4-5) by ELISA and for lupus anti-coagulant.Results: Comparison of LIA with ELISA revealed a good agreement for the consensus criteria aPL aβ2GPI and aCL IgG (kappa = 0.69, 0.68, respectively) and a moderate agreement for IgM (kappa = 0.52, 0.49, respectively). Regarding ELISA, aD1/aD4-5 demonstrated the best performance of differentiating APS from asymptomatic SARD [area under the curve (AUC): 0.76]. aPG IgG had the best performance by LIA (AUC: 0.72) not significantly different from aD1/aD4-5. There was a good agreement for aPG IgG with aD1/aD4-5 (kappa = 0.71).Conclusions: aD1/aD4-5 (ELISA) and aPG IgG (LIA) differentiate APS from SARD patients. PG appears to interact with β2GPI of APS patients and exposes D1 thereof for disease-specific aPL binding in LIA. [ABSTRACT FROM AUTHOR]

Published
2018
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2. Antiphospholipid antibodies detected by line immunoassay differentiate among patients with antiphospholipid syndrome, with infections and asymptomatic carriers

Author

Philipp von Landenberg, Claudia Grossi, Maria Orietta Borghi, Caterina Bodio, Valentina Somma, Michael Mahler, Pier Luigi Meroni, Dirk Roggenbuck, Thomas Büttner, Francesco Boccellato, Peter Schierack, Katja Hanack, Paolo Macor, Roggenbuck, Dirk, Borghi, Maria Orietta, Somma, Valentina, Büttner, Thoma, Schierack, Peter, Hanack, Katja, Grossi, Claudia, Bodio, Caterina, Macor, Paolo, von Landenberg, Philipp, Boccellato, Francesco, Mahler, Michael, and Meroni, Pier Luigi

Subjects
Adult, Male, medicine.drug_class, Immunology, 030204 cardiovascular system & hematology, Monoclonal antibody, Infections, Asymptomatic, Diagnosis, Differential, Line immunoassay, 03 medical and health sciences, Young Adult, 0302 clinical medicine, Rheumatology, Antiphospholipid syndrome, medicine, Humans, Immunology and Allergy, ddc:610, Institut für Biochemie und Biologie, Aged, 030203 arthritis & rheumatology, Immunoassay, Lupus anticoagulant, Antiphospholipid antibody, medicine.diagnostic_test, biology, business.industry, Beta2 - glycoprotein I, Phospholipid binding proteins, Middle Aged, medicine.disease, Antiphospholipid Syndrome, Phospholipid binding protein, IgG binding, biology.protein, Antibodies, Antiphospholipid, Female, medicine.symptom, Antibody, Mathematisch-Naturwissenschaftliche Fakultät, business, Asymptomatic carrier, Research Article
Abstract

Background Antiphospholipid antibodies (aPL) can be detected in asymptomatic carriers and infectious patients. The aim was to investigate whether a novel line immunoassay (LIA) differentiates between antiphospholipid syndrome (APS) and asymptomatic aPL+ carriers or patients with infectious diseases (infectious diseases controls (IDC)). Methods Sixty-one patients with APS (56 primary, 22/56 with obstetric events only, and 5 secondary), 146 controls including 24 aPL+ asymptomatic carriers and 73 IDC were tested on a novel hydrophobic solid phase coated with cardiolipin (CL), phosphatic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, beta2-glycoprotein I (β2GPI), prothrombin, and annexin V. Samples were also tested by anti-CL and anti-β2GPI ELISAs and for lupus anticoagulant activity. Human monoclonal antibodies (humoAbs) against human β2GPI or PL alone were tested on the same LIA substrates in the absence or presence of human serum, purified human β2GPI or after CL-micelle absorption. Results Comparison of LIA with the aPL-classification assays revealed good agreement for IgG/IgM aß2GPI and aCL. Anti-CL and anti-ß2GPI IgG/IgM reactivity assessed by LIA was significantly higher in patients with APS versus healthy controls and IDCs, as detected by ELISA. IgG binding to CL and ß2GPI in the LIA was significantly lower in aPL+ carriers and Venereal Disease Research Laboratory test (VDRL) + samples than in patients with APS. HumoAb against domain 1 recognized β2GPI bound to the LIA-matrix and in anionic phospholipid (PL) complexes. Absorption with CL micelles abolished the reactivity of a PL-specific humoAb but did not affect the binding of anti-β2GPI humoAbs. Conclusions The LIA and ELISA have good agreement in detecting aPL in APS, but the LIA differentiates patients with APS from infectious patients and asymptomatic carriers, likely through the exposure of domain 1. Electronic supplementary material The online version of this article (doi:10.1186/s13075-016-1018-x) contains supplementary material, which is available to authorized users.

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