title: Molecular cloning and characterization of markers and cytokines for equid myeloid cells.
abstract: The myeloid cell system comprises of monocytes, macrophages (MPhi), 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 MPhi 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.
authors.affiliation: Institute for Zoo and Wildlife Research, Alfred-Kowalke-Str. 17, 10315 Berlin, Germany. f.steinbach@vla.defra.gsi.gov.nk
authors.forename: Falko
authors.initials: F
authors.lastname: Steinbach
authors.forename: Robert
authors.initials: R
authors.lastname: Stark
authors.forename: Sherif
authors.initials: S
authors.lastname: Ibrahim
authors.forename: Eman Abd-El
authors.initials: EA
authors.lastname: Gawad
authors.forename: Hanns
authors.initials: H
authors.lastname: Ludwig
authors.forename: Jakob
authors.initials: J
authors.lastname: Walter
authors.forename: Ulrich
authors.initials: U
authors.lastname: Commandeur
authors.forename: Susanne
authors.initials: S
authors.lastname: Mauel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000235:genetics; D018949:Antigens, CD1 / Q000235:genetics; D015214:Antigens, Differentiation, Myelomonocytic / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D016207:Cytokines / Q000235:genetics*; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005819:Genetic Markers; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D006736:Horses / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D022423:Myeloid Cells / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D000097382:CD68 Molecule
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018949:Antigens, CD1; D015214:Antigens, Differentiation, Myelomonocytic; D016207:Cytokines; D004247:DNA; D005819:Genetic Markers; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011994:Recombinant Proteins; C067980:CD68 antigen, human; D000097382:CD68 Molecule