Production and characterization of monoclonal antibodies raised against BoLA class I antigens |
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| Authors: | J J Letesson P Coppe N Lostrie R Greimers A Depelchin |
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| Affiliation: | 1. Départ. Microbiologie, Fac. Univ. Notre Dame de la Paix, 5000 Namur Belgium;2. Centre d''Immunologie Appliquée à l''Elevage, F.N.D.P., 5000 Namur Belgium;3. Départ. Histologie, F.N.D.P., 5000 Namur Belgium;4. Départ. Anat. Pathologie, Univ. de Liège, 4000 Liège Belgium;1. Department of Obstetrics and Gynecology, Hôpital Antoine-Béclère, Assistance Publique-Hôpitaux de Paris, Clamart, France;2. Université Paris-Saclay, Gif-sur-Yvette, France;3. Département d’Anatomopathologie, Hôpital Necker, Assistance Publique-Hôpitaux de Paris, Paris, France;4. University of Paris, Institut National de la Santé et de la Recherche Médicale (INSERM) Unité Mixte de Recherche (UMR) 1151, Paris, France;5. Département de Foetopathologie, Hôpital Antoine-Béclère, Assistance Publique-Hôpitaux de Paris, Clamart, France;6. Département d’Obstétrique et Gynécologie, Hôpital Kremlin-Bicêtre, Assistance Publique-Hôpitaux de Paris, Le Kremlin-Bicêtre, France;7. University of Paris, INSERM UMR 1163, Paris, France;8. Centre de Recherche des Cordeliers, INSERM, Sorbonne University, Université de Paris, Paris, France;9. Laboratoire d’Histocompatibilité, Hôpital de Nouméa, Nouméa, New Caledonia;10. Laboratoire d’Immunologie et Histocompatibilité, Hôpital Saint Louis, Assistance Publique-Hôpitaux de Paris, Paris, France;11. Université de Paris, INSERM UMR 976, Institut de Recherche Saint Louis, Paris, France;12. Département de Néphrologie et Transplantation Rénale, Hôpital Necker, Assistance Publique-Hôpitaux de Paris, Paris, France;1. CHU de Bordeaux, Laboratoire d’Immunologie et Immunogénétique, Hôpital Pellegrin, Place Amélie Raba Léon, Bordeaux, France;2. Immuno ConcEpT, UMR CNRS 5164, 146 rue Léo Saignat, Bordeaux, France;3. Université de Bordeaux, 146 rue Léo Saignat, Bordeaux, France;4. Leiden University Medical Center, Department of Immunohaematology and Blood transfusion, Albinusdreef 2, 2300RC Leiden, the Netherlands;5. One Lambda, Inc., 21001 Kittridge, Canoga Park, CA, USA;6. Université de Bordeaux, Laboratoire ARNA, 146 rue Léo Saignat, Bordeaux, France;7. INSERM U1212, CNRS UMR 5320, IECB, 2 rue Robert Escarpit, Pessac, France |
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| Abstract: | Monoclonal antibodies (MoAbs) reacting with bovine leukocyte membrane antigens have been prepared by fusion of mouse myeloma cells (SP2/0.Ag.14) and spleen cells of mice immunized with various cell types. Three of these MoAbs detected membrane components showing the typical structure of class I MHC molecules; indeed, immunoprecipitation studies revealed that these components were proteins composed of two subunits of 44,000 and 12,000 daltons apparent molecular weight. The density of these antigens in the cells of various leukocyte lineages was determined by solid phase radioimmunoassay, immunogold staining and cytofluorometry. Their expression seemed similar to that of class I molecules in other species, namely heavy on the mononuclear blood cells and weaker on the neutrophils and platelets. The eosinophils appeared more positive than the neutrophils, while the erythrocytes were negative. Cross-inhibition and sequential immunoprecipitation experiments demonstrated that these MoAbs recognised different epitopes either on a single molecule or on cross-reacting molecules. One antibody appeared to be raised against the monomorphic bovine beta-2-microglobulin, while the two other antibodies detected the heavy chain of polymorphic class I-like products. The authors propose that the BoLA class I polymorphism should be studied by determination of the fixation ratio of the monomorphic anti-beta 2M versus the polymorphic anticlass I antibodies amongst the animals. |
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