FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Journal of veterinary science2023; 24(3); e37; doi: 10.4142/jvs.23007

In vitro effects of monophosphoryl lipid A and Poly I:C combination on equine cells.

Abstract: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objective: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.
© 2023 The Korean Society of Veterinary Science.
Get Full Text
Publication Date: 2023-06-05 PubMed ID: 37271505PubMed Central: PMC10244135DOI: 10.4142/jvs.23007Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research evaluates how the combined use of toll-like receptor (TLR) agonists monophosphoryl lipid A (MPL) and polyinosinic:polycytidylic acid (Poly I:C) can enhance immune responses in horses’ cells without damage, suggesting potential use in vaccine development to prevent infections in equines.

Objective and Methodology

  • The study aimed to assess the combined impact of TLR agonists MPL and Poly I:C on various types of equine cells namely peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs).
  • These cell types were obtained from three mixed breed horses and were treated with MPL, Poly I:C, and a combination of both.
  • Using real-time polymerase chain reaction, the study identified the mRNA expression of numerous molecules involved in the immune response, including interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1).

Findings

  • The combination of MPL and Poly I:C significantly potentiated immunomodulatory responses in equine cells and showed no cytotoxic effects.
  • In PBMCs, the combination led to greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C alone.
  • Similarly, the combined treatment significantly escalated mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs.
  • In BM-MSCs, the treatment combination also significantly increased the mRNA expression of IL-8, MCP-1, and VEGF compared to the application of a single TLR agonist.

Conclusion

  • The study concludes that the combination of MPL and Poly I:C can potentially be used as an adjuvant, a substance that improves the body’s immune response to an antigen, in vaccines. This could assist in preventing infectious diseases in horses by stimulating and enhancing the immune system’s response.

Cite This Article

APA
Lee DH, Lee EB, Seo JP, Ko EJ. (2023). In vitro effects of monophosphoryl lipid A and Poly I:C combination on equine cells. J Vet Sci, 24(3), e37. https://doi.org/10.4142/jvs.23007

Publication

Journal of veterinary science
ISSN: 1976-555X
NlmUniqueID: 100964185
Country: Korea (South)
Language: English
Volume: 24
Issue: 3
Pages: e37
PII: e37

Researcher Affiliations

Lee, Dong-Ha
  • College of Veterinary Medicine and Veterinary Medical Research Institute, Jeju National University, Jeju 63243, Korea.
Lee, Eun-Bee
  • Department of Veterinary Medicine, College of Veterinary Medicine, Jeju National University, Jeju 63243, Korea.
Seo, Jong-Pil
  • Department of Veterinary Medicine, College of Veterinary Medicine, Jeju National University, Jeju 63243, Korea.
Ko, Eun-Ju
  • College of Veterinary Medicine and Veterinary Medical Research Institute, Jeju National University, Jeju 63243, Korea.
  • Department of Veterinary Medicine, College of Veterinary Medicine, Jeju National University, Jeju 63243, Korea. eunju@jejunu.ac.kr.

MeSH Terms

  • Humans
  • Horses / genetics
  • Animals
  • Leukocytes, Mononuclear / metabolism
  • Vascular Endothelial Growth Factor A
  • Interleukin-6
  • Interleukin-12 Subunit p40
  • Interleukin-8
  • Cytokines / genetics
  • Cytokines / metabolism
  • Interferon-gamma
  • Adjuvants, Immunologic / pharmacology
  • RNA, Messenger
  • Poly I

Grant Funding

  • Jeju National University

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 38 references
  1. Garçon N, Van Mechelen M. Recent clinical experience with vaccines using MPL- and QS-21-containing adjuvant systems.. Expert Rev Vaccines 2011 Apr;10(4):471-86.
    pubmed: 21506645doi: 10.1586/erv.11.29google scholar: lookup
  2. Figueiredo MD, Vandenplas ML, Hurley DJ, Moore JN. Differential induction of MyD88- and TRIF-dependent pathways in equine monocytes by Toll-like receptor agonists.. Vet Immunol Immunopathol 2009 Jan 15;127(1-2):125-34.
    pubmed: 19019456doi: 10.1016/j.vetimm.2008.09.028google scholar: lookup
  3. Luchner M, Reinke S, Milicic A. TLR Agonists as Vaccine Adjuvants Targeting Cancer and Infectious Diseases.. Pharmaceutics 2021 Jan 22;13(2).
    pmc: PMC7911620pubmed: 33499143doi: 10.3390/pharmaceutics13020142google scholar: lookup
  4. Campbell JD. Development of the CpG Adjuvant 1018: A Case Study.. Methods Mol Biol 2017;1494:15-27.
    pubmed: 27718183doi: 10.1007/978-1-4939-6445-1_2google scholar: lookup
  5. Thompson KA, Strayer DR, Salvato PD, Thompson CE, Klimas N, Molavi A, Hamill AK, Zheng Z, Ventura D, Carter WA. Results of a double-blind placebo-controlled study of the double-stranded RNA drug polyI:polyC12U in the treatment of HIV infection.. Eur J Clin Microbiol Infect Dis 1996 Jul;15(7):580-7.
    pubmed: 8874076doi: 10.1007/bf01709367google scholar: lookup
  6. Gupta SK, Bajwa P, Deb R, Chellappa MM, Dey S. Flagellin a toll-like receptor 5 agonist as an adjuvant in chicken vaccines.. Clin Vaccine Immunol 2014 Mar;21(3):261-70.
    pmc: PMC3957660pubmed: 24451328doi: 10.1128/cvi.00669-13google scholar: lookup
  7. Ziegler A, Gerber V, Marti E. In vitro effects of the toll-like receptor agonists monophosphoryl lipid A and CpG-rich oligonucleotides on cytokine production by equine cells.. Vet J 2017 Jan;219:6-11.
    pubmed: 28093114doi: 10.1016/j.tvjl.2016.11.013google scholar: lookup
  8. Liu T, Nerren J, Murrell J, Juillard V, Garch HE, Martens R. CpG-induced stimulation of cytokine expression by peripheral blood mononuclear cells of foals and their dams. J Equine Vet Sci 2008;28(7):419–426.
  9. Ziegler A, Olzhausen J, Hamza E, Stojiljkovic A, Stoffel MH, Garbani M, Rhyner C, Marti E. An allergen-fused dendritic cell-binding peptide enhances in vitro proliferation of equine T-cells and cytokine production.. Vet Immunol Immunopathol 2022 Jan;243:110351.
    pubmed: 34800874doi: 10.1016/j.vetimm.2021.110351google scholar: lookup
  10. Cassano JM, Schnabel LV, Goodale MB, Fortier LA. The immunomodulatory function of equine MSCs is enhanced by priming through an inflammatory microenvironment or TLR3 ligand.. Vet Immunol Immunopathol 2018 Jan;195:33-39.
    pubmed: 29249315doi: 10.1016/j.vetimm.2017.10.003google scholar: lookup
  11. Pezzanite LM, Chow L, Johnson V, Griffenhagen GM, Goodrich L, Dow S. Toll-like receptor activation of equine mesenchymal stromal cells to enhance antibacterial activity and immunomodulatory cytokine secretion.. Vet Surg 2021 May;50(4):858-871.
    pubmed: 33797775doi: 10.1111/vsu.13628google scholar: lookup
  12. Felippe MJ. Equine Clinical Immunology. 1st ed. Ames: John Wiley & Sons; 2015. pp. 2–3.
  13. Ko EJ, Lee YT, Lee Y, Kim KH, Kang SM. Distinct Effects of Monophosphoryl Lipid A, Oligodeoxynucleotide CpG, and Combination Adjuvants on Modulating Innate and Adaptive Immune Responses to Influenza Vaccination.. Immune Netw 2017 Oct;17(5):326-342.
    pmc: PMC5662782pubmed: 29093654doi: 10.4110/in.2017.17.5.326google scholar: lookup
  14. Ahn SY, Le CTT, Ko EJ. Monophosphoryl Lipid A and Poly I:C Combination Adjuvant Promoted Ovalbumin-Specific Cell Mediated Immunity in Mice Model.. Biology (Basel) 2021 Sep 13;10(9).
    pmc: PMC8471534pubmed: 34571785doi: 10.3390/biology10090908google scholar: lookup
  15. Kim SY, Joo HG. Evaluation of adjuvant effects of fucoidan for improving vaccine efficacy.. J Vet Sci 2015;16(2):145-50.
    pmc: PMC4483496pubmed: 25549218doi: 10.4142/jvs.2015.16.2.145google scholar: lookup
  16. Lopez BS, Hurley DJ, Giancola S, Giguère S, Felippe MJB, Hart KA. The effect of age on foal monocyte-derived dendritic cell (MoDC) maturation and function after exposure to killed bacteria.. Vet Immunol Immunopathol 2019 Apr;210:38-45.
    pubmed: 30947978doi: 10.1016/j.vetimm.2018.11.020google scholar: lookup
  17. Ko EJ, Joo HG. Fucoidan enhances the survival and sustains the number of splenic dendritic cells in mouse endotoxemia.. Korean J Physiol Pharmacol 2011 Apr;15(2):89-94.
    pmc: PMC3104203pubmed: 21660148doi: 10.4196/kjpp.2011.15.2.89google scholar: lookup
  18. Seo JP, Tsuzuki N, Haneda S, Yamada K, Furuoka H, Tabata Y, Sasaki N. Osteoinductivity of gelatin/β-tricalcium phosphate sponges loaded with different concentrations of mesenchymal stem cells and bone morphogenetic protein-2 in an equine bone defect model.. Vet Res Commun 2014 Mar;38(1):73-80.
    pubmed: 24442646doi: 10.1007/s11259-013-9587-5google scholar: lookup
  19. Lee DH, Lee EB, Seo JP, Ko EJ. Evaluation of concurrent vaccinations with recombinant canarypox equine influenza virus and inactivated equine herpesvirus vaccines.. J Anim Sci Technol 2022 May;64(3):588-598.
    pmc: PMC9184697pubmed: 35709134doi: 10.5187/jast.2022.e30google scholar: lookup
  20. Beekman L, Tohver T, Léguillette R. Comparison of cytokine mRNA expression in the bronchoalveolar lavage fluid of horses with inflammatory airway disease and bronchoalveolar lavage mastocytosis or neutrophilia using REST software analysis.. J Vet Intern Med 2012 Jan-Feb;26(1):153-61.
    pubmed: 22168153doi: 10.1111/j.1939-1676.2011.00847.xgoogle scholar: lookup
  21. Andreassen SM, Berg LC, Nielsen SS, Kristensen AT, Jacobsen S. mRNA expression of genes involved in inflammation and haemostasis in equine fibroblast-like synoviocytes following exposure to lipopolysaccharide, fibrinogen and thrombin.. BMC Vet Res 2015 Jun 27;11:141.
    pmc: PMC4483216pubmed: 26116380doi: 10.1186/s12917-015-0448-zgoogle scholar: lookup
  22. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001 Dec;25(4):402-8.
    pubmed: 11846609doi: 10.1006/meth.2001.1262google scholar: lookup
  23. Salkowski CA, Detore GR, Vogel SN. Lipopolysaccharide and monophosphoryl lipid A differentially regulate interleukin-12, gamma interferon, and interleukin-10 mRNA production in murine macrophages.. Infect Immun 1997 Aug;65(8):3239-47.
    pmc: PMC175458pubmed: 9234781doi: 10.1128/iai.65.8.3239-3247.1997google scholar: lookup
  24. Dalrymple SA, Lucian LA, Slattery R, McNeil T, Aud DM, Fuchino S, Lee F, Murray R. Interleukin-6-deficient mice are highly susceptible to Listeria monocytogenes infection: correlation with inefficient neutrophilia.. Infect Immun 1995 Jun;63(6):2262-8.
    pmc: PMC173295pubmed: 7768607doi: 10.1128/iai.63.6.2262-2268.1995google scholar: lookup
  25. Kuo TM, Hu CP, Chen YL, Hong MH, Jeng KS, Liang CC, Chen ML, Chang C. HBV replication is significantly reduced by IL-6.. J Biomed Sci 2009 Apr 20;16(1):41.
    pmc: PMC2687430pubmed: 19374779doi: 10.1186/1423-0127-16-41google scholar: lookup
  26. Kindt TJ, Goldsby RA, Osborne BA, Kuby J. Kuby Immunology. 6th ed. Ames: Macmillan; 2007. pp. 375–377.
  27. Le CTT, Ahn SY, Kang SM, Ko EJ. Functional NK Cell Activation by Ovalbumin Immunization with a Monophosphoryl Lipid A and Poly I:C Combination Adjuvant Promoted Dendritic Cell Maturation.. Vaccines (Basel) 2021 Sep 23;9(10).
    pmc: PMC8540815pubmed: 34696169doi: 10.3390/vaccines9101061google scholar: lookup
  28. Neuder LE, Keener JM, Eckert RE, Trujillo JC, Jones SL. Role of p38 MAPK in LPS induced pro-inflammatory cytokine and chemokine gene expression in equine leukocytes.. Vet Immunol Immunopathol 2009 Jun 15;129(3-4):192-9.
    pubmed: 19070370doi: 10.1016/j.vetimm.2008.11.006google scholar: lookup
  29. Sun WC, Moore JN, Hurley DJ, Vandenplas ML, Fortes B, Thompson R, Linden J. Differential modulation of lipopolysaccharide-induced expression of inflammatory genes in equine monocytes through activation of adenosine A2A receptors.. Vet Immunol Immunopathol 2010 Apr 15;134(3-4):169-77.
    pubmed: 19766323doi: 10.1016/j.vetimm.2009.08.018google scholar: lookup
  30. Hellman S, Hjertner B, Morein B, Fossum C. The adjuvant G3 promotes a Th1 polarizing innate immune response in equine PBMC.. Vet Res 2018 Oct 22;49(1):108.
    pmc: PMC6389152pubmed: 30348190doi: 10.1186/s13567-018-0602-2google scholar: lookup
  31. Naumann K, Wehner R, Schwarze A, Petzold C, Schmitz M, Rohayem J. Activation of dendritic cells by the novel Toll-like receptor 3 agonist RGC100.. Clin Dev Immunol 2013;2013:283649.
    pmc: PMC3878805pubmed: 24454470doi: 10.1155/2013/283649google scholar: lookup
  32. Dietze B, Cierpka E, Schäfer M, Schill W, Lutz MB. An improved method to generate equine dendritic cells from peripheral blood mononuclear cells: divergent maturation programs by IL-4 and LPS.. Immunobiology 2008;213(9-10):751-8.
    pubmed: 18926290doi: 10.1016/j.imbio.2008.07.024google scholar: lookup
  33. Auray G, Facci MR, van Kessel J, Buchanan R, Babiuk LA, Gerdts V. Differential activation and maturation of two porcine DC populations following TLR ligand stimulation.. Mol Immunol 2010 Jul;47(11-12):2103-11.
    pubmed: 20462636doi: 10.1016/j.molimm.2010.03.016google scholar: lookup
  34. Roßmann L, Bagola K, Stephen T, Gerards AL, Walber B, Ullrich A, Schülke S, Kamp C, Spreitzer I, Hasan M, David-Watine B, Shorte SL, Bastian M, van Zandbergen G. Distinct single-component adjuvants steer human DC-mediated T-cell polarization via Toll-like receptor signaling toward a potent antiviral immune response.. Proc Natl Acad Sci U S A 2021 Sep 28;118(39).
    pmc: PMC8488681pubmed: 34561306doi: 10.1073/pnas.2103651118google scholar: lookup
  35. DelaRosa O, Lombardo E. Modulation of adult mesenchymal stem cells activity by toll-like receptors: implications on therapeutic potential.. Mediators Inflamm 2010;2010:865601.
    pmc: PMC2902124pubmed: 20628526doi: 10.1155/2010/865601google scholar: lookup
  36. Salcedo R, Ponce ML, Young HA, Wasserman K, Ward JM, Kleinman HK, Oppenheim JJ, Murphy WJ. Human endothelial cells express CCR2 and respond to MCP-1: direct role of MCP-1 in angiogenesis and tumor progression.. Blood 2000 Jul 1;96(1):34-40.
    pubmed: 10891427
  37. Whelan DS, Caplice NM, Clover AJP. Mesenchymal stromal cell derived CCL2 is required for accelerated wound healing.. Sci Rep 2020 Feb 14;10(1):2642.
    pmc: PMC7021763pubmed: 32060374doi: 10.1038/s41598-020-59174-1google scholar: lookup
  38. Han Y, Yang J, Fang J, Zhou Y, Candi E, Wang J, Hua D, Shao C, Shi Y. The secretion profile of mesenchymal stem cells and potential applications in treating human diseases.. Signal Transduct Target Ther 2022 Mar 21;7(1):92.
    pmc: PMC8935608pubmed: 35314676doi: 10.1038/s41392-022-00932-0google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,824 horse owners like you who receive our email updates on horse health and nutrition.