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.
Publication Date: 2005-08-23 PubMed ID: 16112744DOI: 10.1016/j.vetimm.2005.07.015Google Scholar: Lookup
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- Journal Article
Summary
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The study focuses on the myeloid cell system of equines which includes horses. The researchers cloned specific markers and cytokines imperative for the identification and differentiation of these cells. By doing so, they’ve improved our understanding of how equine immune cells like macrophages and dendritic cells can be characterized.
Understanding the Equine Myeloid Cell System
- The myeloid cell system in equines and other animals consists of certain types of cells that are part of the body’s immune response. This system includes monocytes, macrophages, dendritic cells, Kupffer cells, osteoclasts and microglia. Their differentiation and activation are governed by specific cytokines like GM-CSF or IL-4.
Cloning of Equine Cytokines and Markers
- Although there are markers like CD1, CD14, CD68, CD163, and CD206 that help in the identification and characterization of these cells, they had not been cloned and characterized in equines before this study.
- To overcome this knowledge gap, the study employed two primary approaches – the cloning of equine cytokines and markers and analysis of the cross-reactivity of human homologues or anti-human monoclonal antibodies.
- The researchers used degenerate primers from other species, or equine-specific primers based on information from Genbank for the cloning process.
Characterizing Cell Markers Using Flow Cytometry
- The researchers used flow cytometry to understand the expression of these markers on myeloid cells.
- Cross-reactivity, meaning these markers’ ability to react with related substances, was revealed for anti-human CD14, CD163 and the mannose receptor (CD206) monoclonal antibodies.
Identifying Surface Markers
- Surface markers like CD1 and CD68 that distinguish macrophages and dendritic cells were cloned and sequenced.
- The results clarified the identity of equine CD1a and CD1b through blast homology, meaning they have the comparative analysis of biological sequences.
Impact and Further Application
- The critical outcome of this study was the advancement in characterization methods for equine immune cells such as dendritic cells and macrophages.
- The information gathered can aid in developing treatments or therapies concerning the immune response of horses and potentially other equids.
Cite This Article
APA
Steinbach F, Stark R, Ibrahim S, Gawad EA, Ludwig H, Walter J, Commandeur U, Mauel S.
(2005).
Molecular cloning and characterization of markers and cytokines for equid myeloid cells.
Vet Immunol Immunopathol, 108(1-2), 227-236.
https://doi.org/10.1016/j.vetimm.2005.07.015 Publication
Researcher Affiliations
- Institute for Zoo and Wildlife Research, Alfred-Kowalke-Str. 17, 10315 Berlin, Germany. f.steinbach@vla.defra.gsi.gov.nk
MeSH Terms
- Animals
- Antibodies, Monoclonal
- Antigens, CD / genetics
- Antigens, CD1 / genetics
- Antigens, Differentiation, Myelomonocytic / genetics
- Base Sequence
- Cloning, Molecular
- Cross Reactions
- Cytokines / genetics
- DNA / genetics
- Gene Expression
- Genetic Markers
- Granulocyte-Macrophage Colony-Stimulating Factor / genetics
- Horses / genetics
- Horses / immunology
- Humans
- Myeloid Cells / immunology
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
Citations
This article has been cited 8 times.- Kang H, Lee GKC, Bienzle D, Arroyo LG, Sears W, Lillie BN, Beeler-Marfisi J. Equine alveolar macrophages and monocyte-derived macrophages respond differently to an inflammatory stimulus.. PLoS One 2023;18(3):e0282738.
- Satué K, Gardon JC, Muñoz A. A review of current knowledge of myeloproliferative disorders in the horse.. Acta Vet Scand 2021 Feb 23;63(1):8.
- Saini S, Singha H, Siwach P, Tripathi BN. Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.. Vet World 2019;12(4):496-503.
- Schwab UE, Tallmadge RL, Matychak MB, Felippe MJB. Effects of autologous stromal cells and cytokines on differentiation of equine bone marrow-derived progenitor cells.. Am J Vet Res 2017 Oct;78(10):1215-1228.
- Rőszer T. Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms.. Mediators Inflamm 2015;2015:816460.
- Zamani F, Zare Shahneh F, Aghebati-Maleki L, Baradaran B. Induction of CD14 Expression and Differentiation to Monocytes or Mature Macrophages in Promyelocytic Cell Lines: New Approach.. Adv Pharm Bull 2013;3(2):329-32.
- Cavatorta DJ, Erb HN, Felippe MJ. Activation-induced FoxP3 expression regulates cytokine production in conventional T cells stimulated with autologous dendritic cells.. Clin Vaccine Immunol 2012 Oct;19(10):1583-92.
- Mauel S, Steinbach F, Ludwig H. Monocyte-derived dendritic cells from horses differ from dendritic cells of humans and mice.. Immunology 2006 Apr;117(4):463-73.
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