Immunologic characterization and functional properties of murine antibodies raised against deleted mutants of human beta 2-glycoprotein I.
beta 2-Glycoprotein I (beta 2GPI) is a 50 kDa molecule proposed as a principal target of 'autoimmune' antiphospholipid antibodies (aPL). We have used deleted mutants (DM) representing different domains of beta 2GPI (I-IV, IV-V and V) for immunization of naive mice and studied the characteristics of the respective murine IgG preparations in comparison with affinity-purified IgG from two patients with primary antiphospholipid syndrome. Immunization with beta 2GPI and with the DM produced anti-beta 2GPI antibodies, part of which reacted with negatively charged phospholipids (PL), whereas reactivity with cardiolipin was evident only in the IgG from mice immunized with beta 2GPI. These results are consistent with the presumption that aPL are induced following the in vivo association of beta 2GPI (used for immunization) with resident negatively charged PL. Accordingly, DM which either lack the PL binding site or aPL attachment locus did not elicit, upon immunization, antibodies reactive with PL. Further, murine anti-beta 2GPI IgG and human 'autoimmune' aPL were similar, albeit not identical, in terms of DM requirement for PL binding and charge dependency. Murine antibodies and human aPL, regardless of their binding characteristics, were found to bind significantly to platelets upon their activation with thrombin and to promote platelet activation. The results of the current study emphasize the dissimilarities between human 'autoimmune' aPL and murine anti-beta 2GPI. Thus, anti-beta 2GPI antibodies to different DM as well as human aPL are capable of binding and activating human platelets provided beta 2GPI is present.
George
J
J
Department of Medicine B, Sheba Medical Center, Tel Hashomer, Sackler Faculty of Medicine, Tel-Aviv University, Israel.
Blank
M
M
Gilburd
B
B
Hojnik
M
M
Shenkman
B
B
Tamarin
I
I
Varon
D
D
Matsuura
E
E
Koike
T
T
Shoenfeld
Y
Y
D000818:Animals; D017152:Antibodies, Antiphospholipid / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D016736:Antiphospholipid Syndrome / Q000276:immunology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D001792:Blood Platelets / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D009994:Osmolar Concentration; D019007:P-Selectin / Q000096:biosynthesis; D015539:Platelet Activation / Q000276:immunology; D017384:Sequence Deletion / Q000235:genetics* / Q000276:immunology*; D013917:Thrombin / Q000494:pharmacology; D053482:beta 2-Glycoprotein I
D017152:Antibodies, Antiphospholipid; D006023:Glycoproteins; D007074:Immunoglobulin G; D019007:P-Selectin; D053482:beta 2-Glycoprotein I; D013917:Thrombin