Molecular cloning and characterization of markers and cytokines for equid myeloid cells

The myeloid cell system comprises of monocytes, macrophages (MΦ), dendritic cells (DC), Kupffer cells, osteoclasts or microglia and is also known as the mononuclear phagocytic system (MPS). Essential cytokines to differentiate or activate these cells include GM-CSF or IL-4. Important markers for cha...

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Bibliographic Details
Published in:Veterinary Immunology and Immunopathology 2005-10, Vol.108 (1), p.227-236
Main Authors: Steinbach, Falko, Stark, Robert, Ibrahim, Sherif, Gawad, Eman Abd-El, Ludwig, Hanns, Walter, Jakob, Commandeur, Ulrich, Mauel, Susanne
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal
Antigens, CD - genetics
Antigens, CD1 - genetics
Antigens, Differentiation, Myelomonocytic - genetics
Base Sequence
Cloning
Cloning, Molecular
Cross Reactions
Cytokines
Cytokines - genetics
DNA - genetics
Equine
Gene Expression
Genetic Markers
Granulocyte-Macrophage Colony-Stimulating Factor - genetics
Horses - genetics
Horses - immunology
Humans
Macrophage
Monocyte
Myeloid Cells - immunology
Recombinant Proteins - genetics
Recombinant Proteins - immunology
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Summary:The myeloid cell system comprises of monocytes, macrophages (MΦ), dendritic cells (DC), Kupffer cells, osteoclasts or microglia and is also known as the mononuclear phagocytic system (MPS). Essential cytokines to differentiate or activate these cells include GM-CSF or IL-4. Important markers for characterization include CD1, CD14, CD68, CD163 and CD206. All these markers, however, were not cloned or further characterized in equids by use of monoclonal antibodies earlier. To overcome this problem with the present study, two approaches were used. First, we cloned equine cytokines and markers, and second we analyzed cross-reactivity of human homologues or anti-human monoclonal antibodies. For cloning of equine cytokines and markers, we used degenerate primers delineated from other species, or equine-specific primers based on previous information in Genbank. Flow cytometry was used to determine the expression of markers on myeloid cells. Cross-reactivity could be shown for anti-human CD14, CD163 and mannose receptor (CD206) mAbs. Surface markers such as CD1 and CD68 that distinguish MΦ and DC were cloned and sequenced. According to blast homology, equine CD1a and CD1b could be identified and distinguished. With the resulting information, dendritic cells and macrophages of horses may be characterized.
ISSN:0165-2427
1873-2534
1365-2567
DOI:10.1016/j.vetimm.2005.07.015