Development and characterization of mouse monoclonal antibodies to eight human complement components: Analysis of reactivity with orthologs of nine mammalian genera |
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| Institution: | 1. Unidad de Inmunología Microbiana, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain;2. Experimental Hepatology and Drug Targeting (HEVEFARM), University of Salamanca, IBSAL, CIBERehd, Salamanca, Spain;3. VISAVET Health Surveillance Centre, Universidad Complutense de Madrid, Madrid, Spain;1. Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA). Paraje Arroyo Seco S/N, Tandil, Buenos Aires, Argentina;2. Centro de Investigación Veterinaria de Tandil (CIVETAN)-CONICET, Paraje Arroyo Seco S/N, Tandil, Buenos Aires, Argentina;3. Laboratorio de Virología, Departamento de Producción Animal, Instituto Nacional de Tecnología Agropecuaria (INTA). Ruta 226, Km 73.5, Balcarce, Buenos Aires, Argentina;4. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Rivadavia 1917, Buenos Aires, Argentina;1. Department of Animal Management, College of Veterinary Science and Medicine, Central Luzon State University, Science City of Muñoz 3120, Nueva Ecija, Philippines;2. Biosafety and Environment Section, Philippine Carabao Center National Headquarters and Gene Pool, Science City of Muñoz 3120, Nueva Ecija, Philippines;3. Department of Biological Sciences, College of Arts and Sciences, Central Luzon State University, Science City of Muñoz 3120, Nueva Ecija, Philippines;4. Department of Animal Science, College of Agriculture, Central Luzon State University, Science City of Muñoz 3120, Nueva Ecija, Philippines;1. School of Agriculture and Biology, Shanghai Jiaotong University, 200240, Shanghai, PR China;2. Department of Emergency, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 200240, Shanghai, PR China;3. Tongren Hospital, Shanghai Jiaotong University School of Medicine, 200240, Shanghai, PR China;4. Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai Jiaotong University, 200240, Shanghai, PR China |
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| 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. |
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| Keywords: | Monoclonal antibodies Mammalian complement Antibody specificity |
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