The anti-beta2-glycoprotein I activity in human anti-phospholipid syndrome sera is due to monoreactive low-affinity autoantibodies directed to epitopes located on native beta2-glycoprotein I and preserved during species' evolution.
To characterize the reactivity pattern of Abs directed to beta2-glycoprotein I (anti-beta2GPI) in patients with anti-phospholipid syndrome, we have purified anti-beta2GPI Abs by affinity chromatography using the IgG fractions from sera of five different anti-phospholipid syndrome patients. Affinity-purified anti-beta2GPI were shown to be representative of Abs found in human sera because their activity could be virtually abolished from the IgG preparations after repeated absorptions on immobilized human beta2GPI column. Our results show that affinity-purified anti-beta2GPI: 1) do react with beta2GPI in the absence of any phospholipid, as demonstrated by the lack of phosphorus contaminant in the employed reagents, as well as by their comparable binding activity before and after extensive delipidation procedure; 2) can recognize beta2GPI regardless of its origin from different animal species; 3) are able to bind soluble beta2GPI with a mean Kd value of 4.65 x 10(-6) M (range 3, 4-7, 2 x 10(-6) M); 4) significantly enhance their binding avidity when beta2GPI is linked to a solid support; and 5) appear to be mainly monoreactive autoantibodies. In conclusion, we have shown that human polyclonal anti-beta2GPI are low affinity, mainly monoreactive autoantibodies directed to an epitope located on native beta2GPI, preserved along the species evolution.
Tincani
A
A
Clinical Immunology Unit, Spedali Civili, Brescia, Italy.
Spatola
L
L
Prati
E
E
Allegri
F
F
Ferremi
P
P
Cattaneo
R
R
Meroni
P
P
Balestrieri
G
G
D000293:Adolescent; D000328:Adult; D000915:Antibody Affinity; D000918:Antibody Specificity; D016736:Antiphospholipid Syndrome / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D018604:Epitope Mapping; D005260:Female; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D013045:Species Specificity; D053482:beta 2-Glycoprotein I
D001323:Autoantibodies; D006023:Glycoproteins; D053482:beta 2-Glycoprotein I