IL-4 induced CD23 (FcepsilonRII) up-regulation in equine peripheral blood mononuclear cells and pulmonary alveolar macrophages.
Abstract: The objectives of this study were to quantify the induction of equine CD23 transcripts in equine peripheral blood mononuclear cells (PBMCs) and pulmonary alveolar macrophages cultured with recombinant equine IL-4 (rEq IL-4). PBMCs were isolated from blood drawn from four healthy horses. Bronchoalveolar lavage (BAL) fluid was collected from three healthy horses and alveolar macrophages were purified using adherence to plastic for 120 min. PBMCs and alveolar macrophages were cultured using four different conditions: rEq IL-4 and LPS, LPS alone, rEq IL-4 alone and a media control. Total RNA was isolated from cells cultured for 24 or 48 h. Reverse transcribed mRNA was amplified and quantified in real-time polymerase chain reaction (RT PCR) using a fluorescein labeled internal TaqMan probe for CD23 expression. Without exception, the relative value for CD23 mRNA transcripts from equine PBMCs and pulmonary alveolar macrophages cultured with rEq IL-4 for 24 and 48 h were higher than those cultured with LPS alone or the untreated control. Furthermore, morphologic changes were noted in alveolar macrophages cultured with rEq IL-4 prompting an investigation of cytokine expression levels. Alveolar macrophages cultured with LPS exhibited increased IL-8 and IL-12 p40 expression when compared to rEq IL-4, rEq IL-4 + LPS or the untreated control. These findings support two conclusions, (1) equine CD23 has a role in IL-4 mediated immune responses in the horse and (2) rEq IL-4 can modulate LPS-induced, pro-inflammatory cytokine production by equine pulmonary alveolar macrophages.
Publication Date: 2004-09-08 PubMed ID: 15350754DOI: 10.1016/j.vetimm.2004.05.001Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study tested the response of certain horse immune cells to the introduction of a protein called IL-4, revealing that it increases the production of CD23 transcripts in these cells and likely helps shape immune responses. It also shows the protein’s potential influence on inflammation.
Objective and methodology
- The study aimed to measure the production of CD23 transcripts in certain immune cells in horses, including peripheral blood mononuclear cells (PBMCs) and pulmonary alveolar macrophages, when exposed to a protein called IL-4.
- Immune cells were harvested from four healthy horses. The PBMCs were obtained from their blood, while the alveolar macrophages were collected from bronchoalveolar lavage fluid, which is a method used to examine the cells and particles in the respiratory tract.
- The laboratory experiment exposed these immune cells to varying conditions, featuring different combinations of IL-4 and lipopolysaccharides (LPS), a kind of molecule that provokes immune responses.
- The researchers analyzed the cells at the 24-hour and 48-hour mark, observing the expression of CD23 via real-time PCR, a technique used to amplify and simultaneously quantify targeted DNA molecules.
Findings
- In all cases, it was found that cells exposed to IL-4 produced more CD23 mRNA transcripts compared to those only exposed to LPS or those left untreated.
- Visual observation of the alveolar macrophages showed that IL-4 visibly changed their morphology.
- Looking into cytokine expression levels, the researchers found that macrophages exposed to LPS had increased levels of IL-8 and IL-12 p40 – pro-inflammatory cytokines – compared to cells exposed to IL-4 or both IL-4 and LPS, or those left untreated.
Conclusions
- These observations indicate that the CD23 protein in horses likely plays a part in shaping immune responses that are mediated by IL-4.
- The study also suggests that IL-4 can potentially alter the production of pro-inflammatory cytokines initiated by LPS in equine pulmonary alveolar macrophages, suggesting a role in influencing inflammation.
Cite This Article
APA
Jackson KA, Stott JL, Horohov DW, Watson JL.
(2004).
IL-4 induced CD23 (FcepsilonRII) up-regulation in equine peripheral blood mononuclear cells and pulmonary alveolar macrophages.
Vet Immunol Immunopathol, 101(3-4), 243-250.
https://doi.org/10.1016/j.vetimm.2004.05.001 Publication
Researcher Affiliations
- Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Cell Culture Techniques
- Horses / blood
- Horses / immunology
- Interleukin-4 / immunology
- Leukocytes, Mononuclear / cytology
- Leukocytes, Mononuclear / immunology
- Lipopolysaccharides / immunology
- Lipopolysaccharides / pharmacology
- Macrophages, Alveolar / cytology
- Macrophages, Alveolar / immunology
- Microscopy, Phase-Contrast / veterinary
- RNA, Messenger / biosynthesis
- RNA, Messenger / blood
- RNA, Messenger / genetics
- Receptors, IgE / biosynthesis
- Receptors, IgE / genetics
- Receptors, IgE / immunology
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Up-Regulation / 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.
- Simões J, Batista M, Tilley P. The Immune Mechanisms of Severe Equine Asthma-Current Understanding and What Is Missing. Animals (Basel) 2022 Mar 16;12(6).
- Montero MC, Del Campo M, Bono M, Simon MV, Guerrero J, Lagos N. Neosaxitoxin Inhibits the Expression of Inflammation Markers of the M1 Phenotype in Macrophages. Mar Drugs 2020 May 27;18(6).
- Rőszer T. Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms. Mediators Inflamm 2015;2015:816460.
- Moran G, Folch H, Henriquez C, Ortloff A, Barria M. Reaginic antibodies from horses with recurrent airway obstruction produce mast cell stimulation. Vet Res Commun 2012 Dec;36(4):251-8.
- Morán G, Folch H, Araya O, Burgos R, Barria M. Detection of reaginic antibodies against Faenia rectivirgula from the serum of horses affected with Recurrent Airway Obstruction by an in vitro bioassay. Vet Res Commun 2010 Dec;34(8):719-26.
- Morán G, Burgos R, Araya O, Folch H. In vitro bioassay to detect reaginic antibodies from the serum of horses affected with recurrent airway obstruction. Vet Res Commun 2010 Jan;34(1):91-9.
- Allen CA, Payne SL, Harville M, Cohen N, Russell KE. Validation of quantitative polymerase chain reaction assays for measuring cytokine expression in equine macrophages. J Immunol Methods 2007 Dec 1;328(1-2):59-69.
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