Development and characterization of mouse monoclonal antibodies to eight human complement components: analysis of reactivity with orthologs of nine mammalian genera
Comparative Immunology, Microbiology and Infectious Diseases publica este artículo de investigación
3 de mayo de 2019
Abstract
To study complement function in mammalian leishmanioses, we developed mouse monoclonal antibodies to the human complement system components C1q, C4, factor D, factor H, factor B, properdin, C5 and C9. Antibody specificity was determined by indirect and capture ELISA and by Western blot. In flow cytometry analysis, seven antibodies recognized the cognate component on human serum-opsonized Leishmania promastigotes. Antibody reactivity was screened against promastigotes opsonized with sera of nine mammalian genera: pig, guinea pig, goat, rabbit, cat, dog, hamster, jird and rat. No antibody recognized jird epitopes on promastigotes. Anti-C4, -properdin, and -C5b reacted with the orthologous protein of all other mammals tested except cat (anti-properdin) and hamster (anti-C5b); anti-C9 only recognized the rabbit ortholog, and anti-C1q, -factor B and -factor H did not react with any of the nine orthologs. Such interspecies crossreactive antibodies can be valuable tools for analysis of mammalian complement function in infectious diseases.
Abbreviations
CPclassical complement pathwayLPlectin complement pathwayAPalternative complement pathwayc-ELISAcapture ELISAfBfactor BfDfactor DfHfactor HfIfactor Ii-ELISAindirect ELISAmAbmonoclonal antibodyMBLmannan binding lectin
Keywords
monoclonal antibodiesmammalian complementantibody specificityLeishmania promastigote
Monedero M., Infantes-Lorenzo JA., Moreno I., Martin AB., Macias RIR., Torano A. y Dominguez M.
Servicio de Inmunología. Centro Nacional de Microbiología (CNM). Instituto de Salud Carlos III (ISCIII). | |
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD). Instituto de Salud Carlos III (ISCIII). | |
Instituto de Investigación Biomédica de Salamanca (IBSAL). | |
Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM). Facultad de Farmacia. Universidad de Salamanca (USAL). | |
Servicio de Micobacterias (MYC). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM). | |