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PE-Cy5.5 conjugates bind to the cells expressing mouse DEC205/CD205

Authors
 Chae Gyu Park  ;  Anthony Rodriguez  ;  Ralph M. Steinman 
Citation
 JOURNAL OF IMMUNOLOGICAL METHODS, Vol.384(1-2) : 184-190, 2012 
Journal Title
 JOURNAL OF IMMUNOLOGICAL METHODS 
ISSN
 0022-1759 
Issue Date
2012
MeSH
Animals ; Antibodies, Monoclonal/immunology ; Antibodies, Monoclonal/metabolism ; Antigens, CD/genetics ; Antigens, CD/immunology* ; Antigens, CD/metabolism ; Binding, Competitive/immunology ; CHO Cells ; Carbocyanines/chemistry ; Cricetinae ; Cricetulus ; Dendritic Cells/immunology ; Flow Cytometry ; Gene Expression ; HEK293 Cells ; Humans ; Hybridomas ; Immunoconjugates/chemistry ; Immunoconjugates/immunology* ; Immunoconjugates/metabolism ; Lectins, C-Type/genetics ; Lectins, C-Type/immunology* ; Lectins, C-Type/metabolism ; Mice ; Mice, Inbred C57BL ; Minor Histocompatibility Antigens ; Protein Binding/immunology ; Receptors, Cell Surface/genetics ; Receptors, Cell Surface/immunology* ; Receptors, Cell Surface/metabolism ; Species Specificity ; Streptavidin/chemistry
Keywords
CD205 ; DEC205 ; PE-Cy5.5 ; Nonspecific binding ; Dendritic cells
Abstract
DEC205/CD205, an endocytic receptor of C-type multilectin, is expressed highly in dendritic cells (DCs). DEC205 was shown to efficiently deliver vaccine antigens in surrogate ligands to the antigen processing and presentation machinery of DCs, which resulted in the development of DC-targeted vaccines employing anti-DC monoclonal antibodies (mAbs). During our studies to characterize a variety of anti-DC mAbs including anti-DEC205 by flow cytometric analysis, we discovered that a secondary anti-immunoglobulin antibody conjugated with PE-Cy5.5 bound strongly to the cells expressing mouse DEC205 (mDEC205) without incubation of a primary anti-mDEC205 mAb. In the present study we demonstrate that various antibodies and streptavidin conjugated with PE-Cy5.5 bind to the mDEC205-expressing cells including CHO, KIT6, and HEK293 cells. The interaction between the PE-Cy5.5 conjugates and the cells expressing mDEC205 appears distinctive, since none of the PE-Cy5.5 conjugates bind to the cells that express human DEC205 on surface. Besides, only PE-Cy5.5 conjugates bind strongly to mDEC205-expressing cells; PerCP-Cy5.5, APC-Cy5.5, and Cy5.5 conjugates bind weakly; PE, PE-Cy5, Cy5, FITC, or Alexa488 conjugates do not bind to mDEC205‐expressing cells. Therefore the use of PE-Cy5.5 conjugates, widely utilized in multicolor flow cytometry, requires precaution against nonspecific binding to mDEC205-positive cells.
Full Text
http://www.sciencedirect.com/science/article/pii/S0022175912002050
DOI
22841832
Appears in Collections:
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
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/90848
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