title: Characterization of monoclonal antibodies against porcine pulmonary alveolar macrophages of gnotobiotic miniature swine.
abstract: There is no sufficient porcine specific antibody available to investigate the ontogeny and development of porcine pulmonary alveolar macrophage (PAM). Therefore, several mouse anti-porcine macrophage mAbs have been produced and characterized in this study. First, the monoclonal antibody PM18-7, an IgG1, kappa isotype, bound to 91% of PAM, 6% of monocytes, and 2% of granulocytes indicating PM18-7 was found to be PAM specific. Monocyte derived macrophages could not be induced to express the PM18-7 antigen by culture. PM18-7 was immunoprecipitated with a molecule of 260 kDa under non-reducing conditions and with that of 130 kDa under reducing conditions in SDS-PAGE. Second, the monoclonal antibody PM3-15, an IgG1, kappa isotype, bound to 92% of PAM, 86% of monocytes, and 3% of granulocytes indicating PM3-15 was mononuclear phagocyte specific. PM3-15 was immunoprecipitated with a molecule of 245 kDa under non-reducing conditions and those of 150, 95 kDa under reducing conditions in SDS-PAGE. Third, the monoclonal antibody PM16-6, an IgM isotype, bound to 82% of PAM, 89% of monocytes, and 82% of granulocytes indicating PM16-6 recognizes those cells of myeloid lineage such as macrophages, monocytes and granulocyte. The antigen immunoprecipitated by PM16-6 was 120 kDa under non-reducing conditions and was 75, 25 kDa under reducing conditions. Finally, the antigen bound to PM18-7 was identified as ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1, CD203a), PM3-15 was figured out as integrin alpha M beta 2 precursor (ITGaM, CD11b) and PM16-6 was identified as Thimet oligopeptidase (THOP-1) by the LC-MS/MS protein sequencing method.
authors.affiliation: Department of Bioscience and Biotechnology, Bio-Organ Research Center, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea; Department of Animal Biotechnology, Animal Science, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Jeong Ho
authors.initials: JH
authors.lastname: Hwang
authors.affiliation: Department of Animal Biotechnology, Animal Science, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Sang Eun
authors.initials: SE
authors.lastname: Kim
authors.affiliation: Department of Bioscience and Biotechnology, Bio-Organ Research Center, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Sung Han
authors.initials: SH
authors.lastname: Jung
authors.affiliation: Department of Bioscience and Biotechnology, Bio-Organ Research Center, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Young Kyu
authors.initials: YK
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, University of Health Science and Medicine, Chicago Medical School, North Chicago, IL 60064, USA.
authors.forename: Yoon Berm
authors.initials: YB
authors.lastname: Kim
authors.affiliation: Department of Bioscience and Biotechnology, Bio-Organ Research Center, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea; Department of Animal Biotechnology, Animal Science, Konkuk University, 1-Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea. Electronic address: htl3675@konkuk.ac.kr.
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000032:analysis / Q000276:immunology; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000276:immunology; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006801:Humans; D008264:Macrophages / Q000276:immunology; D016676:Macrophages, Alveolar / Q000166:cytology / Q000276:immunology*; D051379:Mice; D010586:Phagocytes / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine; D013556:Swine, Miniature / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D003165:Complement System Proteins
keywords: Gnotobiotic miniature swine; Monoclonal antibody; Porcine macrophage