re PAMP andnot a constituent of host cell [28]. Since unmethylated CpG motifs are present in the vector DNA, increased mRNA for TLR9  was observed. However, vaccine DNA transfected cells did not show increase in TLR 9 could be due to FMDV VP1 antigen if contributed for the down regulation of TLR 9 in FMDV infected cells, may be the same antigen carried by the DNA vaccine reduced expression of TLR 9 in cells but not significantly. Because in vivo replication of virus lead to more antigen expression in FMDV infected cells. Other possibility could be most RNA viruses have evolved strategies to sequester dsRNA by a variety of mechanisms to avoid activation of these antiviral pathways [29] and down regulation of TLRs might have important immunosuppressive effect [30] or viral persistence which is also observed in FMDV after the natural infection [25].

Our investigation showed very interesting results, showing TLR 2 is up regulated in FMDV infection and FMD DNA vaccine transfection conditions which contribute the antigen. However, major difference of up regulation of TLR 3 & 4, down regulation of TLR 9 observed in FMDV infection needs more studies to support the claim made in this report.

5. Acknowledgements

We are grateful to Department of Biotechnology for the financial support. The authors thank the Director, IVRI and the Joint Director for the facilities provided for carrying out this work.


  1. D. Werling, J. Piercy and T. J. Coffey, “Expression of TollLike Receptors (TLR) by Bovine Antigen-Presenting CellsPotential Role in Pathogen Discrimination?” Veterinary Immunology and Immunopathology, Vol. 112, No. 1-2, 2006, pp. 2-11. doi:10.1016/j.vetimm.2006.03.007
  2. P. V. Barnett and H. Carabin, “A Review of Emergency Foot-and-Mouth Disease (FMD) Vaccines,” Vaccine, Vol. 20, No. 11-12, 2002, pp. 1505-1514. doi:10.1016/S0264-410X(01)00503-5
  3. R. C. Rigden, C. P. Carrasco, P. V. Barnett, A. Summerfield and K. C. McCullough, “Innate Immune Responses Following Emergency Vaccination against Foot-andMouth Disease Virus in Pigs,” Vaccine, Vol. 21, No. 13-14, 2003, pp. 1466-1477. doi:10.1016/S0264-410X(02)00663-1
  4. A. L. Barnard, A. Arriens, S. Cox, P. Barnett, B. Kristensen, A. Summerfield and K. C. McCullough, “Immune Response Characteristics Following Emergency Vaccination of Pigs against Foot-and-Mouth Disease,” Vaccine, Vol. 23, No. 8, 2005, pp. 1037-1047. doi:10.1016/j.vaccine.2004.07.034
  5. Q. Yao, P. Qian, Q. Huang, Y. Cao and H. Chen, “Comparison of Immune Responses to Different Foot-andMouth Disease Genetically Engineered Vaccines in Guinea Pigs,” Journal of Virological Methods, Vol. 147, No. 1, 2008, pp. 143-150. doi:10.1016/j.jviromet.2007.08.027
  6. D. Dory, M. Remond, V. Beven, R. Cariolet, S. Zientara and A. Jestin, “Foot-and-Mouth Disease Virus neutralizing Antibodies Production Induced by pcDNA3 and Sindbis Virus Based Plasmid Encoding FMDV P1-2A3C3D in Swine,” Antiviral Research, Vol. 83, No. 1, 2009, pp. 45- 52. doi:10.1016/j.antiviral.2009.03.004
  7. A. K. Chockalingam, S. Thiyagarajan, N. Govindasamy, R. Patnaikuni, S. Garlapati, R. R. Golla, D. H. Joyappa, P. Krishnamshetty and V. V. Veluvarti, “Study of a Chimeric Foot-and-Mouth Disease Virus DNA Vaccine Containing Structural Genes of Serotype O in a Genome Backbone of Serotype Asia 1 in Guinea pigs,” Acta Virologica, Vol. 54, No. 3, 2010, pp. 189-195. doi:10.4149/av_2010_03_189
  8. A. Summerfield, L. Guzylack-Piriou, L. Harwood and K. C. McCullough, “Innate Immune Responses against Footand-Mouth Disease Virus: Current Understanding and Future Directions,” Veterianry Immunology and Immunopathology, Vol. 128, No. 1-3, 2009, pp. 205-210. doi:10.1016/j.vetimm.2008.10.296
  9. J. Sambrook and D. W. Russell, “The Condensed Protocols from Molecular Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2006.
  10. A. Boyum, “Isolation of Mononuclear Cells and Granulocytes from Human Blood. Isolation of Monuclear Cells by One Centrifugation, and of Granulocytes by Combining Centrifugation and Sedimentation at 1 g,” Scandinavian Journal of Clinical & Laboratory Investigation, Vol. 97, 1968, pp. 77-89.
  11. S. A. Altman, L. Randers and G. Rao, “Comparison of Trypan Blue Dye Exclusion and Fluorometric Assays for Mammalian Cell Viability Determinations,” Biotechnology Progress, Vol. 9, No. 6, 1993, pp. 671-674. doi:10.1021/bp00024a017
  12. P. Konstantinova, O. Brake, J. Haasnoot, P. De Haan and B. Berkhout, “Trans-Inhibition of HIV-1 by a Long Hairpin RNA Expressed within the Viral Genome,” Retrovirology, Vol. 4, 2007, p. 15. doi:10.1186/1742-4690-4-15
  13. K. W. Boehme, M. Guerrero and T. Compton, “Human Cytomegalovirus Envelope Glycoproteins B and H Are Necessary for TLR2 Activation in Permissive Cells,” Journal of Immunology, Vol. 177, No. 10, 2006, pp. 7094-7102.
  14. G. M. Barton, “Viral Recognition by Toll-Like Receptors,” Seminars in Immunology, Vol. 19, No. 1, 2007, pp. 33-40. doi:10.1016/j.smim.2007.01.003
  15. Y. Sato, M. Roman, H. Tighe, D. Lee, M. Corr, M. D. Nguyen, G. J. Silverman, M. Lotz, D. A. Carson and E. Raz, “Immunostimulatory DNA Sequences Necessary for Effective Intradermal Gene Immunization,” Science, Vol. 273, No. 5273, 1996, pp. 352-354. doi:10.1126/science.273.5273.352
  16. D. M. Klinman, G. Yamshchikov and Y. Ishigatsubo, “Contribution of CpG Motifs to the Immunogenicity of DNA Vaccines,” Journal of Immunology, Vol. 158, No. 8, 1997, pp. 3635-3639.
  17. W. W. Leitner, H. Ying and N. P. Restifo, “DNA and RNA-Based Vaccines: Principles, Progress and Prospects,” Vaccine, Vol. 18, No. 9-10, 1999, pp. 765-777. doi:10.1016/S0264-410X(99)00271-6
  18. E. Pfaff, H. J. Thiel, E. Beck, K. Strohmaier and H. Schaller, “Analysis of Neutralizing Epitopes on Foot-andMouth Disease Virus,” Journal of Virology, Vol. 62, No. 6, 1988, pp. 2033-2040.
  19. M. Leippert, E. Beck, F. Weiland and E. Pfaff, “Point Mutations within the Beta G - Beta H Loop of Foot-andMouth Disease Virus O1K Affect Virus Attachment to Target Cells,” Journal of Virology, Vol. 71, No. 2, 1997, pp. 1046-1051.
  20. S. B. Nagendrakumar, G. S. Reddy, D. Chandran, D. Thiagarajan, P. N. Rangarajan and V. A. Srinivasan, “Molecular Characterization of Foot-and-Mouth Disease Virus Type C of Indian Origin,” Journal of Clinical Microbiology, Vol. 43, No. 2, 2005, pp. 966-969. doi:10.1128/JCM.43.2.966-969.2005
  21. A. Rudensky, P. Preston-Hurlburt, S. C. Hong, A. Barlow and C. A. Janeway Jr., “Sequence Analysis of Peptides Bound to MHC Class II Molecules,” Nature, Vol. 353, No. 6345, 1991, pp. 622-627. doi:10.1038/353622a0
  22. A. A. Andersen, “Cross Reactions of Normal Bovine Sera with Foot-and-Mouth Disease Virus: Incidence, Duration, and Effect of Shipping Stress,” American Journal of Veterinary Research, Vol. 39, No. 4, 1978, pp. 603-606.
  23. K. Tabeta, P. Georgel, E. Janssen, X. Du, K. Hoebe, K. Crozat, S. Mudd, L. Shamel, S. Sovath, J. Goode, L. Alexopoulou, R. A. Flavell and B. Beutler, “Toll-Like Receptors 9 and 3 as Essential Components of Innate immune Defense against Mouse Cytomegalovirus Infection,” Proceedings of the National Academy of Sciences USA, Vol. 101, No. 10, 2004, pp. 3516-3521. doi:10.1073/pnas.0400525101
  24. K. Crozat and B. Beutler, “TLR7: A New Sensor of Viral Infection,” Proceedings of the National Academy of Sciences USA, Vol. 101, No. 18, 2004, pp. 6835-6836. doi:10.1073/pnas.0401347101
  25. Z. Zhang, J. B. Bashiruddin, C. Doel, J. Horsington, S. Durand and S. Alexandersen, “Cytokine and Toll-Like Receptor mRNAs in the Nasal-Associated Lymphoid Tissues of Cattle during Foot-and-Mouth Disease Virus Infection,” Journal of Comparitive Pathology, Vol. 134, No. 1, 2006, pp. 56-62. doi:10.1016/j.jcpa.2005.06.011
  26. U. Zahringer, B. Lindner, S. Inamura, H. Heine and C. Alexander, “TLR2-Promiscuous or Specific? A Critical Re-Evaluation of a Receptor Expressing Apparent Broad Specificity,” Immunobiology, Vol. 213, No. 3-4, 2008, pp. 205-224. doi:10.1016/j.imbio.2008.02.005
  27. G. Cheng, X. Zhao, W. Yan, W. Wang, X. Zuo, K. Huang, Y. Liu, J. Chen, J. Wang, W. Cong, M. Liu, H. Gao, Y. Lu and Z. Zheng, “Alpha Interferon Is a Powerful Adjuvant for a Recombinant Protein Vaccine against Footand-Mouth Disease Virus in Swine, and an Effective Stimulus of in Vivo Immune Response,” Vaccine, Vol. 25, No. 28, 2007, pp. 5199-5208. doi:10.1016/j.vaccine.2007.04.089
  28. E. Peterhans, T. W. Jungi and M. Schweizer, “BVDV and Innate Immunity,” Biologicals, Vol. 31, No. 2, 2003, pp. 107-112. doi:10.1016/S1045-1056(03)00024-1
  29. M. Schroder and A. G. Bowie, “An Arms Race: Innate Antiviral Responses and Counteracting Viral Strategies,” Biochemical Society Transactions, Vol. 35, No. 6, 2007, pp. 1512-1514. doi:10.1042/BST0351512
  30. M. V. Borca, I. Gudmundsdottir, I. J. Fernandez-Sainz, L. G. Holinka and G. R. Risatti, “Patterns of Cellular Gene Expression in Swine Macrophages Infected with Highly Virulent Classical Swine Fever Virus Strain Brescia,” Virus Research, Vol. 138, No. 1-2, 2008, pp. 89-96. doi:10.1016/j.virusres.2008.08.009


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