Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.
Abstract: Interleukin (IL)-4 and IL-10 activate plethora of immune cells and induce the humoral immune response. However, recombinant version of horse IL-4 and IL-10 has not been investigated to understand their immunomodulating activities. This study aimed to produce recombinant horse mature IL-4 and IL-10 in . Immune-modulating activities of recombinant horse IL-4 and IL-10 were investigated in peripheral blood mononuclear cells (PBMCs). Methods: Equine PBMCs were stimulated with recombinant IL-4 and IL-10. A proliferation of PBMCs was measured by XTT assay and cytokines induction was measured by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Results: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis displayed a molecular weight of 15 kDa for IL-4 and 19 kDa for IL-10. Recombinant IL-4 and IL-10 significantly induced cell proliferation at 250 ng/ml. The results demonstrated that IL-4 enhanced expression of interferon-gamma (IFN-γ), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-10, while recombinant horse IL-10 induced expression of IL-6, IFN-γ, and TNF-α. Conclusions: The present study demonstrated that biologically active horse IL-4 and IL-10 could be produced in .
Publication Date: 2019-04-03 PubMed ID: 31190704PubMed Central: PMC6515821DOI: 10.14202/vetworld.2019.496-503Google Scholar: Lookup
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- Journal Article
- Biochemistry
- Cell Proliferation
- Cells
- Cytokines
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Enzymes
- Equine Diseases
- Equine Health
- Genetics
- Immune Response
- Immune System
- In Vitro Research
- Inflammation
- Interleukins
- Molecular biology
- Polymerase Chain Reaction
- Real-Time PCR
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
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This research focuses on the production and examination of the immune-modulating activities of lab-produced horse interleukin-4 and interleukin-10 (IL-4 and IL-10), two types of immune cells. The researchers found that these cells caused a mixed inflammatory cytokine response in peripheral blood mononuclear cells, indicating their potential use in immunomodulatory therapy.
Methods
- The study started with the production of recombinant horse IL-4 and IL-10.
- These cells were then introduced to peripheral blood mononuclear cells (PBMCs), a type of immune cell found in the bloodstream,
- After stimulation of the PBMCs with these recombinant IL-4 and IL-10, a few measurements were taken:
- A proliferation (increase in number) of PBMCs was noted using an XTT assay, a test used to measure cell viability and proliferation.
- The induction of cytokines, small proteins that are crucial in cell signaling during immune responses, was measured using an enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR).
Results
- After analyzing the samples using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the researchers found that the molecular weights of IL-4 and IL-10 were 15 and 19 kDa respectively.
- More importantly, the researchers discovered that both the lab-produced IL-4 and IL-10 significantly induced cell proliferation at a concentration of 250 ng/ml.
- On a more specific level, IL-4 was found to increase the expression of interferon-gamma (IFN-γ), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-10. Meanwhile, the lab-produced horse IL-10 induced expression of IL-6, IFN-γ, and TNF-α.
Conclusion
- The research successfully showed that biologically active horse IL-4 and IL-10 could be produced in a lab setting and are able to stimulate immune reactions in horse peripheral blood mononuclear cells.
- This finding may have important implications for the development of immunomodulatory medicine, specifically for horses. However, further research is needed to ascertain the potential therapeutic uses and understand any possible side effects.
Cite This Article
APA
Saini S, Singha H, Siwach P, Tripathi BN.
(2019).
Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.
Vet World, 12(4), 496-503.
https://doi.org/10.14202/vetworld.2019.496-503 Publication
Researcher Affiliations
- ICAR-National Research Centre on Equines, Hisar, Haryana, India.
- Department of Biotechnology, Chaudhary Devi Lal University, Sirsa, Haryana, India.
- ICAR-National Research Centre on Equines, Hisar, Haryana, India.
- Department of Biotechnology, Chaudhary Devi Lal University, Sirsa, Haryana, India.
- ICAR-National Research Centre on Equines, Hisar, Haryana, India.
References
This article includes 32 references
- Mitchell RE, Hassan M, Burton BR, Britton G, Hill EV, Verhagen J, Wraith DC. IL-4 enhances IL-10 production in Th1 cells: implications for Th1 and Th2 regulation.. Sci Rep 2017 Sep 12;7(1):11315.
- Wynn TA. Type 2 cytokines: mechanisms and therapeutic strategies.. Nat Rev Immunol 2015 May;15(5):271-82.
- Hsieh CS, Heimberger AB, Gold JS, O'Garra A, Murphy KM. Differential regulation of T helper phenotype development by interleukins 4 and 10 in an alpha beta T-cell-receptor transgenic system.. Proc Natl Acad Sci U S A 1992 Jul 1;89(13):6065-9.
- McLeod JJ, Baker B, Ryan JJ. Mast cell production and response to IL-4 and IL-13.. Cytokine 2015 Sep;75(1):57-61.
- Junttila IS. Tuning the Cytokine Responses: An Update on Interleukin (IL)-4 and IL-13 Receptor Complexes.. Front Immunol 2018;9:888.
- Rojas JM, Avia M, Martín V, Sevilla N. IL-10: A Multifunctional Cytokine in Viral Infections.. J Immunol Res 2017;2017:6104054.
- MacKenzie KF, Pattison MJ, Arthur JS. Transcriptional regulation of IL-10 and its cell-specific role in vivo.. Crit Rev Immunol 2014;34(4):315-45.
- Cordeau ME, Joubert P, Dewachi O, Hamid Q, Lavoie JP. IL-4, IL-5 and IFN-gamma mRNA expression in pulmonary lymphocytes in equine heaves.. Vet Immunol Immunopathol 2004 Jan;97(1-2):87-96.
- Curto E, Messenger KM, Salmon JH, Gilger BC. Cytokine and chemokine profiles of aqueous humor and serum in horses with uveitis measured using multiplex bead immunoassay analysis.. Vet Immunol Immunopathol 2016 Dec;182:43-51.
- Pacholewska A, Jagannathan V, Drögemüller M, Klukowska-Rötzler J, Lanz S, Hamza E, Dermitzakis ET, Marti E, Leeb T, Gerber V. Impaired Cell Cycle Regulation in a Natural Equine Model of Asthma.. PLoS One 2015;10(8):e0136103.
- Steinbach F, Stark R, Ibrahim S, Gawad EA, Ludwig H, Walter J, Commandeur U, Mauel S. Molecular cloning and characterization of markers and cytokines for equid myeloid cells.. Vet Immunol Immunopathol 2005 Oct 18;108(1-2):227-36.
- Dohmann K, Wagner B, Horohov DW, Leibold W. Expression and characterisation of equine interleukin 2 and interleukin 4.. Vet Immunol Immunopathol 2000 Dec 29;77(3-4):243-56.
- Wagner B, Robeson J, McCracken M, Wattrang E, Antczak DF. Horse cytokine/IgG fusion proteins--mammalian expression of biologically active cytokines and a system to verify antibody specificity to equine cytokines.. Vet Immunol Immunopathol 2005 May 1;105(1-2):1-14.
- Berghaus LJ, Giguère S, Bordin AI, Cohen ND. Effects of priming with cytokines on intracellular survival and replication of Rhodococcus equi in equine macrophages.. Cytokine 2018 Feb;102:7-11.
- Wagner B, Perkins G, Babasyan S, Freer H, Keggan A, Goodman LB, Glaser A, Torsteinsdóttir S, Svansson V, Björnsdóttir S. Neonatal Immunization with a Single IL-4/Antigen Dose Induces Increased Antibody Responses after Challenge Infection with Equine Herpesvirus Type 1 (EHV-1) at Weanling Age.. PLoS One 2017;12(1):e0169072.
- Singha H, Goyal SK, Malik P. Cloning and sequence analysis of interleukin (IL)-2, IL-4,IL-10, and IL-18 of Marwari horse.. Indian J. Anim. Sci. 2015;85(6):621–627.
- Sambrook J.J, Green M.R. Molecular Cloning:A Laboratory Manual. New York: 4. Cold Spring Harbor Laboratory Press; 2012. pp. 1599–1609.
- Figueiredo MD, Salter CE, Andrietti AL, Vandenplas ML, Hurley DJ, Moore JN. Validation of a reliable set of primer pairs for measuring gene expression by real-time quantitative RT-PCR in equine leukocytes.. Vet Immunol Immunopathol 2009 Sep 15;131(1-2):65-72.
- Sánchez-Matamoros A, Kukielka D, De las Heras AI, Sánchez-Vizcaíno JM. Development and evaluation of a SYBR Green real-time RT-PCR assay for evaluation of cytokine gene expression in horse.. Cytokine 2013 Jan;61(1):50-3.
- Overbergh L, Valckx D, Waer M, Mathieu C. Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR.. Cytokine 1999 Apr;11(4):305-12.
- Cao S, Liu J, Song L, Ma X. The protooncogene c-Maf is an essential transcription factor for IL-10 gene expression in macrophages.. J Immunol 2005 Mar 15;174(6):3484-92.
- Morris SC, Orekhova T, Meadows MJ, Heidorn SM, Yang J, Finkelman FD. IL-4 induces in vivo production of IFN-gamma by NK and NKT cells.. J Immunol 2006 May 1;176(9):5299-305.
- Oliver JA, Stolberg VR, Chensue SW, King PD. IL-4 acts as a potent stimulator of IFN-γ expression in CD8+ T cells through STAT6-dependent and independent induction of Eomesodermin and T-bet.. Cytokine 2012 Jan;57(1):191-9.
- Lavoie-Lamoureux A, Moran K, Beauchamp G, Mauel S, Steinbach F, Lefebvre-Lavoie J, Martin JG, Lavoie JP. IL-4 activates equine neutrophils and induces a mixed inflammatory cytokine expression profile with enhanced neutrophil chemotactic mediator release ex vivo.. Am J Physiol Lung Cell Mol Physiol 2010 Oct;299(4):L472-82.
- Hochrein H, O'Keeffe M, Luft T, Vandenabeele S, Grumont RJ, Maraskovsky E, Shortman K. Interleukin (IL)-4 is a major regulatory cytokine governing bioactive IL-12 production by mouse and human dendritic cells.. J Exp Med 2000 Sep 18;192(6):823-33.
- Cush SS, Reynoso GV, Kamenyeva O, Bennink JR, Yewdell JW, Hickman HD. Locally Produced IL-10 Limits Cutaneous Vaccinia Virus Spread.. PLoS Pathog 2016 Mar;12(3):e1005493.
- Ma X, Yan W, Zheng H, Du Q, Zhang L, Ban Y, Li N, Wei F. Regulation of IL-10 and IL-12 production and function in macrophages and dendritic cells.. F1000Res 2015;4.
- Fiorentino DF, Zlotnik A, Mosmann TR, Howard M, O'Garra A. IL-10 inhibits cytokine production by activated macrophages.. J Immunol 1991 Dec 1;147(11):3815-22.
- D'Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells.. J Exp Med 1993 Sep 1;178(3):1041-8.
- Stordeur P, Poulin LF, Craciun L, Zhou L, Schandené L, de Lavareille A, Goriely S, Goldman M. Cytokine mRNA quantification by real-time PCR.. J Immunol Methods 2002 Jan 1;259(1-2):55-64.
- Godornes C, Leader BT, Molini BJ, Centurion-Lara A, Lukehart SA. Quantitation of rabbit cytokine mRNA by real-time RT-PCR.. Cytokine 2007 Apr;38(1):1-7.
- Seko Y, Cole S, Kasprzak W, Shapiro BA, Ragheb JA. The role of cytokine mRNA stability in the pathogenesis of autoimmune disease.. Autoimmun Rev 2006 May;5(5):299-305.
Citations
This article has been cited 1 times.- Benítez-Páez A, Gómez Del Pugar EM, López-Almela I, Moya-Pérez Á, Codoñer-Franch P, Sanz Y. Depletion of Blautia Species in the Microbiota of Obese Children Relates to Intestinal Inflammation and Metabolic Phenotype Worsening.. mSystems 2020 Mar 24;5(2).
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