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Generation of anti-human DEC205/CD205 monoclonal antibodies that recognize epitopes conserved in different mammals

 Chae Gyu Park  ;  Anthony Rodriguez  ;  Hisashi Ueta  ;  Haekyung Lee  ;  Maggi Pack  ;  Kenjiro Matsuno  ;  Ralph M. Steinman 
 JOURNAL OF IMMUNOLOGICAL METHODS, Vol.377(1-2) : 15-22, 2012 
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Issue Date
Animals ; Antibodies, Monoclonal/biosynthesis* ; Antibodies, Monoclonal/immunology ; Antigens, CD/immunology* ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Dendritic Cells/immunology* ; Epitopes/immunology ; Humans ; Immunohistochemistry ; Lectins, C-Type/immunology* ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Minor Histocompatibility Antigens ; Receptors, Cell Surface/immunology* ; Recombinant Fusion Proteins/immunology ; Specific Pathogen-Free Organisms ; Transfection
Monoclonal antibody ; Conserved epitope ; CD205 ; DEC205 ; Dendritic cells
DEC205/CD205 is a C-type multilectin receptor, expressed highly in dendritic cells (DCs). Previous efforts to generate anti-human DEC205 (anti-hDEC205) monoclonal antibodies (mAbs) from mice immunized with subdomain proteins of hDEC205 resulted in a few mAbs. Recently, we expressed and utilized a full-length extracellular domain protein of hDEC205 to successfully generate 5 strong anti-hDEC205 mAbs from mice. In this study, DEC205 knockout (KO) mice were immunized with this full-length extracellular domain protein of hDEC205. One of the 3 immunized DEC205 KO mice was chosen for the highest anti-hDEC205 titer by flow cytometric analysis of serum samples on CHO cells stably expressing hDEC205 (CHO/hDEC205 cells) and used for hybridoma fusion. From a single fusion, more than 400 anti-hDEC205 hybridomas were identified by flow cytometric screen with CHO/hDEC205 cells, and a total of 115 hybridomas secreting strong anti-hDEC205 mAb were saved and named HD1 through HD115. To characterize in detail, 10 HD mAbs were chosen for superior anti-hDEC205 reactivity and further subjected to cloning and purification. Interestingly, out of those 10 chosen anti-hDEC205 HD mAbs, 5 mAbs were also strongly reactive to mouse DEC205 while 8 mAbs were found to stain DEC205+ DCs on monkey spleen sections. In addition, we also identified that HD83, one of the 10 chosen HD mAbs, stains DEC205+ DCs in rat spleen and lymph node. Therefore, by immunizing DEC205 KO mice with a full-length extracellular domain protein of hDEC205, we generated a large number of strong anti-hDEC205 mAbs many of which are cross-species reactive and able to visualize DEC205+ DCs in lymphoid tissues of other mammals.
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1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
Yonsei Authors
Park, Chae Gyu(박채규) ORCID logo https://orcid.org/0000-0003-1906-1308
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