A soluble recombinant multimeric anti-Rh(D) single-chain Fv/CR1 molecule restores the immune complex binding ability of CR1-deficient erythrocytes.
CR1 (CD35, the C3b/C4b receptor) is a widely distributed membrane glycoprotein with a unique cluster conformation on the surface of erythrocytes (E). CR1 on E is responsible for the transport of immune complexes (IC) to liver and spleen. As a cofactor of the C3b cleavage by factor I, CR1 is also a potent inhibitor of C activation and inflammation. In some diseases (systemic lupus erythematosus, hemolytic anemia, AIDS, etc.) an acquired low level of CR1 on E has been observed, leading to an impaired clearance of IC. The aim of this study was to design a heterofunctional molecule that will bind to E and restore a normal or a supranormal CR1 density on E that could mimic the unique distribution pattern of CR1 on normal E. For that purpose a new multimerizing system based on the properties of the C-terminal part of the alpha-chain of the C4 binding protein (C4bp) was used. We first produced a multimeric soluble CR1 that proved to be a better inhibitor of in vitro C activation than the monomeric form of CR1, then a heteromultimeric molecule made of CR1 and single-chain Fv anti-Rh(D) valences able to attach E and providing E with as much as a 10-fold increase in CR1 density with the same CR1 distribution pattern as native E. CR1/single-chain Fv anti-Rh(D)-treated E were able in vitro to attach as many opsonized IC as native E. These data open the way for future use of multimeric and heteromultimeric forms of soluble recombinant CR1 as therapy of IC diseases.
Oudin
S
S
Centre Hospitalier Universitaire Reims, Laboratoire d'Immunologie, Unité de Formation et de Recherche Médecine Université de Reims Champugne Ardennes, Pôle Biomolécules IFR53 Reims, France.
Libyh
M T
MT
Goossens
D
D
Dervillez
X
X
Philbert
F
F
Réveil
B
B
Bougy
F
F
Tabary
T
T
Rouger
P
P
Klatzmann
D
D
Cohen
J H
JH
D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D016466:CHO Cells / Q000378:metabolism; D002461:Cell Line, Transformed; D003169:Complement Inactivator Proteins / Q000494:pharmacology; D006224:Cricetinae; D004912:Erythrocytes / Q000276:immunology* / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D007518:Isoantibodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D017463:Receptors, Complement 3b / Q000037:antagonists & inhibitors / Q000172:deficiency* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D012204:Rh-Hr Blood-Group System / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D018029:Rho(D) Immune Globulin; D012995:Solubility
D000936:Antigen-Antibody Complex; D003169:Complement Inactivator Proteins; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D007518:Isoantibodies; C061961:RHO(D) antibody; D017463:Receptors, Complement 3b; D011994:Recombinant Proteins; D012204:Rh-Hr Blood-Group System; D018029:Rho(D) Immune Globulin; C026410:Rho(D) antigen; C055966:immunoglobulin Fv