title: Immunization of a rabbit with beta 2-glycoprotein I induces charge-dependent crossreactive antibodies that bind anionic phospholipids and have similar reactivity as autoimmune anti-phospholipid antibodies.
abstract: A rabbit immunized with beta 2-glycoprotein I (beta 2-GPI) produced Abs that bind to negatively charged phospholipids and to beta 2-GPI. After affinity purification of the Abs to beta 2-GPI, the dual reactivity could still be detected. Adsorption studies with a phosphatidylserine affinity column depleted phospholipid-reactive Abs, but beta 2-GPI reactivity was retained. The same pattern of reactivity was found with culture supernatants from rabbit anti-beta 2-GPI splenocytes fused with an immortalized rabbit cell line. The reactivity to negatively charged phospholipids is likely to involve ionic interactions, as high ionic strength buffers eliminated binding to anionic phospholipids, but not to beta 2-GPI. Affinity-purified anti-phospholipid (aPL) Abs from four of seven autoimmune patients bound anionic phospholipids in the absence of beta 2-GPI. However, in high ionic strength buffer, this binding was abolished in three patients and significantly reduced in the fourth. In contrast, affinity-purified aPL Abs from seven autoimmune patients bound to beta 2-GPI-coated plates, and binding in high ionic strength buffer was reduced only moderately in three patients. Therefore, autoimmune-type aPL Abs display anti-beta 2-GPI reactivity and charge-dependent binding to anionic phospholipids similar to affinity-purified rabbit anti-beta 2-GPI Abs.
authors.affiliation: Department of Immunology, Allergy and Infectious Disease, University of New South Wales, St. George Hospital, Kogarah, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Kouts
authors.forename: M X
authors.initials: MX
authors.lastname: Wang
authors.forename: S
authors.initials: S
authors.lastname: Adelstein
authors.forename: S A
authors.initials: SA
authors.lastname: Krilis
mesh_terms: D000818:Animals; D000838:Anions / Q000276:immunology*; D017152:Antibodies, Antiphospholipid / Q000276:immunology*; D018033:Antibodies, Bispecific / Q000276:immunology*; D000917:Antibody Formation; D002459:Cell Fusion; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003429:Cross Reactions / Q000276:immunology; D006023:Glycoproteins / Q000008:administration & dosage / Q000276:immunology* / Q000302:isolation & purification; D006825:Hybridomas; D007114:Immunization; D007525:Isoelectric Focusing; D008081:Liposomes / Q000276:immunology; D009994:Osmolar Concentration; D010718:Phosphatidylserines / Q000276:immunology; D010743:Phospholipids / Q000276:immunology*; D011817:Rabbits; D053482:beta 2-Glycoprotein I
chemical_list: D000838:Anions; D017152:Antibodies, Antiphospholipid; D018033:Antibodies, Bispecific; D006023:Glycoproteins; D008081:Liposomes; D010718:Phosphatidylserines; D010743:Phospholipids; D053482:beta 2-Glycoprotein I