Characterization of TLR expression in Staphylococcus aureus induced mastitis in mice model by probe based real time PCR

SUSWETA DAS MITRA; SANKAR KUMAR GHOSH; P KRISHNAMOORTHY; ANAMIKA CHAKRABORTY; NIMITA VENUGOPAL C; MANISHA ROY; BIBEK RANJAN SHOME; HABIBUR RAHMAN

doi:10.56093/ijans.v84i10.44237

Authors

SUSWETA DAS MITRA National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
SANKAR KUMAR GHOSH Assam University, Silchar
P KRISHNAMOORTHY National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
ANAMIKA CHAKRABORTY National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
NIMITA VENUGOPAL C National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
MANISHA ROY National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
BIBEK RANJAN SHOME National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024
HABIBUR RAHMAN National Institute of Veterinary Epidemiology and Disease Informatics, Hebbal, Bengaluru, Karnataka 560 024

https://doi.org/10.56093/ijans.v84i10.44237

Keywords:

Bovine mastitis, Intramammary infection, Staphylococcus aureus, Toll like receptors

Abstract

Staphylococcus aureus infection of the bovine mammary gland can result in a spectrum of clinical outcomes ranging from acute to chronic and subclinical. A molecular understanding of the principles causing this is particularly important to eventually develop innovative strategies for prevention and treatment in mastitis. The present study characterized the temporal (2, 4, 8, 12, 24 and 48h) expression of 5 important toll like receptors (TLR 2, TLR 4, TLR 9, TLR 11 and TLR 12) induced by S. aureus intramammary inoculation (IMI) in established mice model employing probe based real time PCR (RT-PCR). S. aureus challenge provoked transcriptional expression of each of the 5 TLRs in a temporal manner with maximum expression at 8 h after IMI. S. aureus maximally altered the mRNA concentration of TLR 9 and TLR 12 (> 5 fold) over the PBS control. We found TLR2 to be weakly (1- to 2- fold) regulated by the S. aureus challenge in this study. This study emphasized the contribution of other TLRs besides the most commonly studied TLR 2 and TLR 4 in activating immune response by switching on several receptors mediated signalling cascades. This study sheds light into the diverse pathogen recognition receptor (PRR) mediated mechanisms involved in host pathogen interaction.

Downloads

Download data is not yet available.

References

1. Akira S, Uematsu S and Takeuchi O. 2006. Pathogen recognition and innate immunity. Cell 124: 783–801. DOI: https://doi.org/10.1016/j.cell.2006.02.015

2. Bannerman D D, Paape M J, Lee J W, Zhao X, Hope J C and Rainard P. 2004. Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clinical and Diagnostic Laboratory Immunology 11: 463–472. DOI: https://doi.org/10.1128/CDLI.11.3.463-472.2004

3. Fournier B and Philpott D J. 2005. Recognition of Staphylococcus aureus by the innate immune system. Clinical Microbiology Review 18: 521–540. DOI: https://doi.org/10.1128/CMR.18.3.521-540.2005

4. Gazzinelli R T and Denkers E Y. 2006. Protozoan encounters with Toll-like receptor signalling pathways: implications for host parasitism. Nature Reviews Immunology 6: 895–906. DOI: https://doi.org/10.1038/nri1978

5. Gerold G, Zychlinsky A, and De Diego J L. 2007. What is the role of Toll-like receptors in bacterial infections? Seminars in Immunology 19: 41–47. DOI: https://doi.org/10.1016/j.smim.2006.12.003

6. Günther J, Esch K, Poschadel N, Petzl W, Zerbe H, Mitterhuemer S, Blum H and Seyfert H M. 2011. Comparative kinetics of Escherichia coli- and Staphylococcus aureus-specific activation of key immune pathways in mammary epithelial cells demonstrates that S. aureus elicits a delayed response dominated by interleukin-6 (IL-6) but not by IL-1A or tumor necrosis factor alpha. Infection and Immunity 79: 695–707. DOI: https://doi.org/10.1128/IAI.01071-10

7. He X, Jing Z and Cheng G. 2014. MicroRNAs: new regulators of Toll-like receptor signalling pathways. BioMed Research International: 945169. DOI: https://doi.org/10.1155/2014/945169

8. Krishnamoorthy P, Satyanarayana, M L, Shome B R and Rahman H. 2014. Mouse milk somatic cell count in coagulase negative Staphylococcus species induced mastitis. Journal of Laboratory Animal Science 2: 16–19.

9. Lee J W, Bannerman D D, Paape M J, Huang M K and Zhao X. 2006. Characterization of cytokine expression in milk somatic cells during intramammary infections with Escherichia coli or Staphylococcus aureus by real-time PCR. Veterinary Research 37: 219–29. DOI: https://doi.org/10.1051/vetres:2005051

10. Menzies M and Ingham A. 2006. Identification and expression of Toll-like receptors 1–10 in selected bovine and ovine tissues. Veterinary Immunology and Immunopathology 109: 23–30. DOI: https://doi.org/10.1016/j.vetimm.2005.06.014

11. Mitra S D, Velu D, Bhuvana M, Krithiga N, Banerjee A, Shome R, Rahman H, Ghosh S K and Shome BR. 2013. Staphylococcus aureus spa type t267, clonal ancestor of bovine subclinical mastitis in India. Journal of Applied Microbiology 114: 1604–15. DOI: https://doi.org/10.1111/jam.12186

12. Modak R, Mitra S D, Krishnamoorthy P, Bhat A, Banerjee A, Gowsica B R, Bhuvana M, Dhanikachalam V, Krithiga N , Shome R, Shome B R and Kundu TK. 2012. Histone H3K14 and H4K8 hyperacetylation is associated with Escherichia coli-induced mastitis in mice. Epigenetics 7: 492– 501. DOI: https://doi.org/10.4161/epi.19742

13. Quinn S R and O’Neill L A. 2011. A trio of microRNAs that control Toll-like receptor signalling. International Immunology 23: 421–25. DOI: https://doi.org/10.1093/intimm/dxr034

14. Shi Z, Cai Z, Yu J, Zhang T, Zhao S, Smeds E, Zhang Q, Wang F, Zhao C, Fu S, Sankar Ghosh, and Dekai Zhang. 2012. Toll- like receptor 11 (TLR11) prevents Salmonella penetration into the murine peyer patches. Journal of Biological Chemistry 287: 43417–23. DOI: https://doi.org/10.1074/jbc.M112.411009

15. Tabeta K, Georgel P, Janssen E, Du X, Hoebe K, Crozat K, Mudd S, Shamel L, Sovath S, Goode J, Alexopoulou L, Flavell R.A and Beutler B. 2004. Toll-like receptors 9 and 3 as essential components of innate immune defense against mouse cytomegalovirus infection. Proceedings of National Academy of Sciences U.S.A. 101:3516–21 DOI: https://doi.org/10.1073/pnas.0400525101

16. Takeuchi O, and Akira S. 2010. Pattern recognition receptors and inflammation. Cell 140 : 805–20. DOI: https://doi.org/10.1016/j.cell.2010.01.022

17. Takeda K, Kaisho T and Akira S. 2003. Toll-like receptors. Annual Review of Immunology 21: 335–76. DOI: https://doi.org/10.1146/annurev.immunol.21.120601.141126

18. Tanaka J, Sugimoto K, Shiraki K, Tameda M, Kusagawa S, Nojiri K, Beppu T, Yoneda K, Yamamoto N, Uchida K, Kojima T and Takei Y. 2010. Functional cell surface expression of Toll-like receptor 9 promotes cell proliferation and survival in human hepatocellular carcinomas. International Journal of Oncology 37: 805–14. DOI: https://doi.org/10.3892/ijo_00000730

19. Wellnitz O and Bruckmaier R M. 2012. The innate immune response of the bovine mammary gland to bacterial infection. Veterinary Journal 192: 148–52. DOI: https://doi.org/10.1016/j.tvjl.2011.09.013

20. Yone V N C, Brelaz M A, Neves J K A L, Ferraz J C, and Pereira V R A. 2012. Role of TNF-alpha, IFN-gamma, and IL-10 in the development of pulmonary tuberculosis. Pulmonary Medicine Volume 2012, ArticleID745483, 10pages doi:10.1155/2012/745483. DOI: https://doi.org/10.1155/2012/745483