Naturally occurring anti-band 3 antibodies bind to protein rather than to carbohydrate on band 3.
ABSTRACT Naturally occurring anti-band 3 antibodies were affinity purified from pooled human IgG (Sandoglobulin) (Lutz, H. U., Flepp, R., and Stringaro-Wipf, G. (1984) J. Immunol. 133, 2610-2618). They bound to the major integral membrane protein of human red blood cells and its 55-kDa NH2-terminal chymotryptic fragment but not to the carbohydrate-rich 38-kDa fragment on blots. Likewise, neither an endo-beta-galactosidase nor a neuraminidase treatment of band 3 on intact red cells reduced their binding to the blotted antigen. Lactoferrin (10 micrograms/ml) had no significant effect on their binding to band 3 and to its 55-kDa chymotryptic fragment. Even in the presence of 20 micrograms/ml lactoferrin anti-band 3 antibodies bound specifically to chymotrypsin-pretreated and oxidatively stressed red cells. Thus, naturally occurring anti-band 3 antibodies bind to protein rather than carbohydrate within band 3 protein, irrespectively of whether the antibodies were depleted of anti-idiotypic and other IgG-reactive antibodies or not.
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Article: Naturally occurring anti-band 3 antibodies bind to protein rather than to carbohydrate on band 3.
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ABSTRACT: Naturally occurring anti-band 3 antibodies (anti-band 3 NAbs) are directed against the 55-kDa chymotryptic fragment of the anion transport protein (band 3) of red blood cells (RBCs). They bind to senescent and oxidatively stressed RBCs and induce their selective clearance. These IgG NAbs exist at low concentrations, and have a weak affinity that prevents them from actively recruiting second binding sites. Cellular senescence or oxidative damage induces a cascade of biochemical events that results in the detachment of band 3 from the cytoskeleton and in clustering of band 3 protein by bound hemichromes and Syk kinase. Clustered band 3 proteins allow bivalent binding of anti-band 3 NAbs. Bivalently bound anti-band 3 NAbs have the unique capacity to stimulate C3b deposition by preferentially generating C3b2-IgG complexes, which act as potent C3 convertase precursors of the alternative complement pathway. Antibody binding not only to clustered, but also to oligomerized band 3 protein further increases if the human plasma also contains induced anti-lactoferrin antibodies. These bind to the polylactosaminyl oligosaccharide, a carbohydrate that exists in lactoferrin and in the 38-kDa fragment of band 3 protein. Anti-lactoferrin antibodies are found primarily in plasma of patients with autoimmune diseases and who have anti-neutrophil cytoplasmic antibodies (ANCA).Transfusion Medicine and Hemotherapy 10/2012; 39(5):321-327. DOI:10.1159/000342171 · 2.01 Impact Factor
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ABSTRACT: This review focuses on the analysis and evaluation of the diverse senescence markers suggested to prime red blood cells (RBC) for clearance in humans. These tags develop in the course of biochemical and structural alterations accompanying RBC aging, as the decrease of activities of multiple enzymes, the gradual accumulation of oxidative damage, the loss of membrane in form of microvesicles, the redistribution of ions and alterations in cell volume, density, and deformability. The actual tags represent the penultimate galactosyl residues, revealed by desialylation of glycophorins, or the aggregates of the anion exchanger (band 3 protein) to which anti-galactose antibodies bind in the first and anti-band 3 naturally occurring antibodies (NAbs) in the second case. While anti-band 3 NAbs bind to the carbohydrate-free portion of band 3 aggregates in healthy humans, induced anti-lactoferrin antibodies bind to the carbohydrate-containing portion of band 3 and along with anti-band 3 NAbs may accelerated clearance of senescent RBC in patients with anti-neutrophil cytoplasmic antibodies (ANCA). Exoplasmically accessible phosphatidylserine (PS) and the alterations in the interplay between CD47 on RBC and its receptor on macrophages, signal regulatory protein alpha (SIRPalpha protein), were also reported to induce erythrocyte clearance. We discuss the relevance of each mechanism and analyze the strength of the data.Frontiers in Physiology 12/2013; 4:387. DOI:10.3389/fphys.2013.00387
Article: Naturally occurring antibodies.[Show abstract] [Hide abstract]
ABSTRACT: Germline-encoded naturally occurring autoantibodies (NAbs) developed about 400 to 450 million years ago to provide specificity for clearance ofbody waste in animals with 3 germ layers. Such NAbs became a necessity to selectively clear aged red blood cells (RBC) surviving 60 to 120 d in higher vertebrates. IgG NAbs to senescent RBC are directed to the most abundant integral membrane protein, the anion-transport protein or band 3 protein, but only bind firmly upon its oligomerization, which facilitates bivalent binding. The main constituent of RBC, the oxygen-carrying hemoglobin, is susceptible to oxidative damage. Oxidized hemoglobin forms hemichromes (a form of aggregates) that bind to the cytoplasmic portion of band 3 protein, induces their clustering on the cytoplasmic, as well as the exoplasmic side and thereby provides the prerequisites for the low affinity IgG anti-band 3 NAbs to bind bivalently. Bound anti-band 3 NAbs overcome their low numbers per RBC by stimulating complement amplification. An affinity for C3 outside the antigen binding region is responsible for a preferential formation of C3b(2)-IgG complexes from anti-band 3 NAbs. These complexes first bind oligomeric properdin, which enhances their affinity for factor B in assembling an alternative C3 convertase.Advances in Experimental Medicine and Biology 01/2012; 750:76-90. DOI:10.1007/978-1-4614-3461-0_6 · 2.01 Impact Factor