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Immunogenetics

, Volume 65, Issue 11, pp 829–834 | Cite as

Breed-linked polymorphisms of porcine toll-like receptor 2 (TLR2) and TLR4 and the primary investigation on their relationship with prevention against Mycoplasma pneumoniae and bacterial LPS challenge

  • Xiaomin Fang
  • Xiao Liu
  • Cui Meng
  • Yanfeng Fu
  • Xuemin Wang
  • Bixia Li
  • Feng Tu
  • Fang Zhao
  • Shouwen RenEmail author
Brief Communication

Abstract

Toll-like receptors (TLRs) play a crucial role in innate immunity, serving as pattern-recognition receptors and the first barrier in host defense against microbial infections. Genetic variations of TLR2 and TLR4 are closely associated with a variety of infectious diseases, particularly lung diseases. In this study, we detected six and four single nucleotide polymorphisms (SNPs) in the coding sequences of porcine TLR2 and TLR4 genes, respectively. Only SNP 1027C>A of TLR4 was shown to be markedly biased in Western and Oriental pig populations. Hence, the susceptibility of pigs with different genotype at position 1027C>A to Mycoplasma hyopneumoniae (Mhp) infection was investigated, and changes to the expression of TLR2, TLR4, TNF-α and IL-1β were monitored. The results showed that there was no significant difference in susceptibility to Mhp infection between AA and CC individuals despite expression levels for all detected genes of the challenge groups being significantly higher than the corresponding control groups. Furthermore, porcine alveolar macrophages of different genotype were collected and stimulated by lipopolysaccharide. We found that the expression of TLR2, TLR4, TNF-α and IL-1β genes were enhanced to different levels by lipopolysaccharide stimulation. TLR2 and TLR4 gene expressions and their rates of increase of 1027CC pigs were significantly higher than for 1027AC pigs (P < 0.01), while TNF-α and IL-1β expressions were significantly lower than for 1027AC pigs (P < 0.01). We predict that allele C at position 1027 of the TLR4 gene contributes to the pig's immune response to gram-negative bacterial infections.

Keywords

Pig Toll-like receptor (TLR) SNPs Mycoplasma pneumoniae LPS 

Notes

Acknowledgments

We thank the Key Laboratory of Veterinary Biological Engineering and Technology for their help with the artificial M. pneumoniae infection experimental work. This work was supported by the Agricultural Science and Technology Innovation Fund Project of Jiangsu Province and the National Swine Industry Technology System, People's Republic of China.

References

  1. Akira S, Takeda K (2004) Toll-like receptor signalling. Nat Rev Immunol 4:499–511PubMedCrossRefGoogle Scholar
  2. Amass SF, Clark LK, van Alstine WG, Bowersock TL, Murphy DA, Knox KE, Albregts SR (1994) Interaction of Mycoplasma hyopneumoniae and Pasteurella multocida infections in swine. J Am Vet Med Assoc 204:102–107PubMedGoogle Scholar
  3. Basta S, Carrasco CP, Knoetig SM, Rigden RC, Gerber H, Summerfield A, McCullough KC (2000) Porcine alveolar macrophages: poor accessory or effective suppressor cells for T-lymphocytes. Veterinary immunology and immunopathology 77:177–190PubMedCrossRefGoogle Scholar
  4. Bell JK, Mullen GE, Leifer CA, Mazzoni A, Davies DR, Segal DM (2003) Leucine-rich repeats and pathogen recognition in Toll-like receptors. Trends in immunology 24:528–533PubMedCrossRefGoogle Scholar
  5. Ben-Ali M, Barbouche M-R, Bousnina S, Chabbou A, Dellagi K (2004) Toll-like receptor 2 Arg677Trp polymorphism is associated with susceptibility to tuberculosis in Tunisian patients. Clinical and diagnostic laboratory immunology 11:625–626PubMedGoogle Scholar
  6. Da Costa CU, Wantia N, Kirschning CJ, Busch DH, Rodriguez N, Wagner H, Miethke T (2004) Heat shock protein 60 from Chlamydia pneumoniae elicits an unusual set of inflammatory responses via Toll-like receptor 2 and 4 in vivo. Eur J Immunol 34:2874–2884PubMedCrossRefGoogle Scholar
  7. Deutscher AT, Jenkins C, Minion FC, Seymour LM, Padula MP, Dixon NE, Walker MJ, Djordjevic SP (2010) Repeat regions R1 and R2 in the P97 paralogue Mhp271 of Mycoplasma hyopneumoniae bind heparin, fibronectin and porcine cilia. Mol Microbiol 78:444–458PubMedCrossRefGoogle Scholar
  8. Eder W, Klimecki W, Yu L, von Mutius E, Riedler J, Braun-Fahrländer C, Nowak D, Martinez FD (2004) Toll-like receptor 2 as a major gene for asthma in children of European farmers. Journal of allergy and clinical immunology 113:482–488PubMedCrossRefGoogle Scholar
  9. Fang X, Xu N, Ren S (2008) Cloning, sequencing and identification of single nucleotide polymorphisms of partial sequence on the porcine CACNA1S gene. Science in China Series C: Life Sciences 51:317–325PubMedCrossRefGoogle Scholar
  10. Hawn TR, Verbon A, Janer M, Zhao LP, Beutler B, Aderem A (2005) Toll-like receptor 4 polymorphisms are associated with resistance to Legionnaires' disease. Proc Natl Acad Sci U S A 102:2487–2489PubMedCrossRefGoogle Scholar
  11. Heine H, Ulmer AJ (2005) Recognition of bacterial products by toll-like receptors. Chem Immunol Allergy 86:99–119PubMedCrossRefGoogle Scholar
  12. Hussein YM, Awad HA, Shalaby SM, Ali A-SA, Alzahrani SS (2012) Toll-like receptor 2 and toll-like receptor 4 polymorphisms and susceptibility to asthma and allergic rhinitis: a case–control analysis. Cell Immunol 274:34–38PubMedCrossRefGoogle Scholar
  13. Into T, Nodasaka Y, Hasebe A, Okuzawa T, Nakamura J-i, Ohata N, Shibata K-i (2002) Mycoplasmal lipoproteins induce toll-like receptor 2-and caspases-mediated cell death in lymphocytes and monocytes. Microbiology and immunology 46:265–276Google Scholar
  14. Liu L, Ma S (2007) Epidemiological characteristics and its prevention of swine enzootic pneumonia. Chinese Journal of Animal Husbandry and Veterinary Medicine 9:64–65Google Scholar
  15. Liu S, Lin W, Shi X, Zhang J, Yao Y, Ming F (2007) Bi-PASA genetic marker and polymorphism of the porcine TLR2 gene. Heilongjiang Animal Science and Veterinary Medicine 1:7–9Google Scholar
  16. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  17. Lorenz E, Mira JP, Cornish KL, Arbour NC, Schwartz DA (2000) A novel polymorphism in the toll-like receptor 2 gene and its potential association with staphylococcal infection. Infection and immunity 68:6398–6401PubMedCrossRefGoogle Scholar
  18. Muneta Y, Uenishi H, Kikuma R, Yoshihara K, Shimoji Y, Yamamoto R, Hamashima N, Yokomizo Y, Mori Y (2003) Porcine TLR2 and TLR6: identification and their involvement in Mycoplasma hyopneumoniae infection. Journal of interferon & cytokine research 23:583–590CrossRefGoogle Scholar
  19. Nicolò C, Di Sante G, Procoli A, Migliara G, Piermattei A, Valentini M, Delogu G, Cittadini A, Constantin G, Ria F (2013) M. tuberculosis in the adjuvant modulates time of appearance of CNS-specific effector T cells in the spleen through a polymorphic site of TLR2. PloS one 8:e55819PubMedCrossRefGoogle Scholar
  20. Shao GQ, Liu MJ, Sun PY, Wang JC, Du G, Zhou Y, Liu D (2007) The establishment of an experimental swine model of swine mycoplasma pneumonia. Journal of Microbes and Infection 2:215–218Google Scholar
  21. Shimizu T KY, Kuwano K (2005) A dipalmitoylated lipoprotein from Mycoplasma pneumoniae activates NF-kappa B through TLR1, TLR2, and TLR6. J Immunol 175:4641–4646Google Scholar
  22. Shinkai H, Tanaka M, Morozumi T, Eguchi-Ogawa T, Okumura N, Muneta Y, Awata T, Uenishi H (2006) Biased distribution of single nucleotide polymorphisms (SNPs) in porcine Toll-like receptor 1 (TLR1), TLR2, TLR4, TLR5, and TLR6 genes. Immunogenetics 58:324–330PubMedCrossRefGoogle Scholar
  23. Xin JC, Wang L, Li Y (2006) Research progress on swine Mycoplasma hyopneumoniae. Swine Industry Science 23:18–22Google Scholar
  24. Yamamoto M, Sato S, Hemmi H, Sanjo H, Uematsu S, Kaisho T, Hoshino K, Takeuchi O, Kobayashi M, Fujita T (2002) Essential role for TIRAP in activation of the signalling cascade shared by TLR2 and TLR4. Nature 420:324–329PubMedCrossRefGoogle Scholar
  25. Yang I, Barton S, Rorke S, Cakebread J, Keith T, Clough J, Holgate S, Holloway J (2004) Toll-like receptor 4 polymorphism and severity of atopy in asthmatics. Genes and immunity 5:41–45PubMedCrossRefGoogle Scholar
  26. Zhang F, Gao XD, Wu WW, Gao Y, Zhang YW, Wang SP (2013) Polymorphisms in toll-like receptors 2, 4 and 5 are associated with Legionella pneumophila infection. Infection 23: (in press)Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Xiaomin Fang
    • 1
  • Xiao Liu
    • 1
  • Cui Meng
    • 1
  • Yanfeng Fu
    • 1
  • Xuemin Wang
    • 1
  • Bixia Li
    • 1
  • Feng Tu
    • 1
  • Fang Zhao
    • 1
  • Shouwen Ren
    • 1
    Email author
  1. 1.Institute of Animal ScienceJiangsu Academy of Agricultural SciencesNanjingPeople’s Republic of China

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