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Sub-lethal Dose of Atropine Gives Protection from Japanese Encephalitis Virus Infection in Chick Embryo Model

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Abstract

Japanese encephalitis (JE) is now a public health concern due to limitation of the available vaccine and nonexistence of specific antiviral therapy and thus demands new potential therapeutical strategies. The present study was conducted to evaluate the effect of atropine in controlling JE virus infection in chick embryo, which is already established as an ideal model for study of various host–pathogen interactions and screening of newer antiviral agents. In this experiment we pre-treated 12 day old embryos with atropine sulphate to see whether it can give protection from JE virus challenge. There were significant decrease in the viral loads in the brain, chorioallantoic membrane and amniotic fluid of the embryo in atropine sulphate pre-treated groups compared to the infection groups. Atropine sulphate pre-treated group showed significant up-regulation of interferon alpha and toll like receptor-3 mRNA, than other analogous groups. The histological changes showed significant reduction of necrosis, inflammation and other co-morbid pathology in the atropine sulphate pre-treated groups. Hence this study demonstrated the protective role of atropine in controlling the JE virus replication and its organ-tropic dissemination to brain which may pave way for a prospective therapy in the future treatment of JE.

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References

  • Adi, A.A.A.M., N.M. Astawa, P.A.A. Damayanti, I.M. Kardena, I.G.M.K. Erawan, I.W. Suardana, I.G.A.A. Putra, and Y. Matsumoto. 2016. Seroepidemiological Evidence for the Presence of Japanese Encephalitis Virus Infection in Ducks, Chickens, and Pigs, Bali-Indonesia. Bali Medical Journal 5(3): 533–537.

    Google Scholar 

  • Anderson, K. 1940. Pathogenesis of Herpes Simplex Virus Infection in Chick Embryos. American Journal of Pathology 6: 137–156.

    Google Scholar 

  • Awais, M., K. Wang, X. Lin, W. Qian, and N. Zhang. 2017. TLR7 Deficiency Leads to TLR8 Compensative Regulation of Immune Response Against JEV in Mice. Frontiers in Immunology 8: 160.

    Article  Google Scholar 

  • Barjesteh, N., S. Behboudi, J.T. Brisbin, A.I. Villanueva, E. Nagy, and S. Sharif. 2014. TLR Ligands Induce Antiviral Responses in Chicken Macrophages. PLoS ONE 9(8): e105713.

    Article  Google Scholar 

  • Baxby, D., and C.J.M. Rondle. 1965. Plaque Formation by Vaccinia Virus in Tissue Cultures Inhibited by 5-Fluorodeoxyuridine. Journal of General Microbiology 38: 423–427.

    Article  CAS  Google Scholar 

  • Biziagos, E., J.M. Crance, J. Passagot, and R. Deloince. 1990. Inhibitory Effects of Atropine, Protamine, and Their Combination on Hepatitis A Virus Replication in PLC/PRF/5 Cells. Antimicrobial Agents and Chemotherapy 34(6): 1112–1117.

    Article  CAS  Google Scholar 

  • Cao, L., S. Fu, X. Gao, M. Li, S. Cui, X. Li, Y. Cao, W. Lei, Z. Lu, Y. He, H. Wang, J. Yan, G.F. Gao, and G. Liang. 2016. Low Protective Efficacy of the Current Japanese Encephalitis Vaccine Against the Emerging Genotype 5 Japanese Encephalitis Virus. PLOS Neglected Tropical Diseases 10(5): e0004686.

    Article  Google Scholar 

  • Chakraborty, U., S. Katoch, M. Sinha, D. Nayak, A. Khurana, and R. Manchanda. 2018. Changes in Viral Load in Different Organs of Japanese Encephalitis Virus-Infected Chick Embryo Under the Influence of belladonna 200C. Indian Journal of Research in Homoeopathy 12: 75–80.

    Article  Google Scholar 

  • Chang, R.Y., T.W. Hsu, Y.L. Chen, S.F. Liu, Y.J. Tsai, Y.T. Lin, Y.S. Chen, and Y.H. Fan. 2013. Japanese Encephalitis Virus Non-coding RNA Inhibits Activation of Interferon by Blocking Nuclear Translocation of Interferon Regulatory Factor 3. Veterinary Microbiology 166: 11–21.

    Article  CAS  Google Scholar 

  • Cleton, N.B., A. Bosco-Lauth, M.J. Page, and R.A. Bowen. 2014. Age-Related Susceptibility to Japanese Encephalitis Virus in Domestic Ducklings and Chicks. American Journal of Tropical Medicine and Hygiene 90(2,5): 242–246.

    Article  Google Scholar 

  • Chomczynski, P., and N. Sacchi. 1987. Single-Step Method of RNA Isolation by Acid Guanidinium Thiocyanate-Phenol-Chloroform Extraction. Analytical Biochemistry 162: 156–159.

    Article  CAS  Google Scholar 

  • Curtis, D.R., and J.W. Phillis. 1960. The Action of Procaine and Atropine on Spinal Neurones. The Journal of Physiology 153: 17–34.

    Article  CAS  Google Scholar 

  • Daffis, S., M.A. Samuel, M.S. Suthar, M. Gale, and M.S. Diamond Jr. 2008. Toll-Like Receptor 3 has a Protective Role Against West Nile Virus Infection. Journal of Virology 82: 10349–10358.

    Article  CAS  Google Scholar 

  • De Elio, F.J. 1948. Acetylcholine Antagonists: A Comparison of Their Action in Differents Tissues. British Journal of Pharmacology and Chemotherapy 3: 108–112.

    Article  Google Scholar 

  • Erecinska, M., D. Nelson, and I.A. Silver. 1988. Interactions of Benztropine, Atropine and Ketamine with Veratridine-Activated Sodium Channels: Effects on Membrane Depolarization, K + - Efflux and Neurotransmitter Amino Acid Release. British Journal of Pharmacology 94: 871–881.

    Article  CAS  Google Scholar 

  • Fan, W., S. Qian, P. Qian, and X. Li. 2016. Antiviral Activity of Luteolin Against Japanese Encephalitis Virus. Virus Research 15(220): 112–116.

    Article  Google Scholar 

  • Gilhare, V.S., S.D. Hirpurkar, A. Kumar, S.K. Naik, and T. Sahu. 2015. Pock Forming Ability of Fowl Pox Virus Isolated from Layer Chicken and Its Adaptation in Chicken Embryo Fibroblast Cell Culture. Veterinary World 8(3): 245–250.

    Article  Google Scholar 

  • Glasel, J. 1995. Validity of Nucleic Acid Purities Monitored by 260 nm/280 nm Absorbance Ratios. BioTechniques 18: 62–63.

    CAS  PubMed  Google Scholar 

  • Han, Y.W., C.J. Youn, E. Uyangaa, S.B. Kim, and J.H. Kim. 2014. Distinct Dictation of Japanese Encephalitis Virus-Induced Neuroinflammation and Lethality via Triggering TLR3 and TLR4 Signal Pathways. PLoS Pathogens 10(9): e1004319.

    Article  Google Scholar 

  • Hiscott, J. 2007. Triggering the Innate Antiviral Response Through IRF-3 Activation. Journal of Biological Chemistry 282: 15325–15329.

    Article  CAS  Google Scholar 

  • Jacobsen, I.D., K. Große, A. Berndt, and B. Hube. 2011. Pathogenesis of Candida Albicans Infections in the Alternative Chorio-Allantoic Membrane Chicken Embryo Model Resembles Systemic Murine Infections. PLoS ONE 6(5): e19741.

    Article  CAS  Google Scholar 

  • Janzer, R.C., and M.C. Raff. 1987. Astrocytes Induce Blood-Brain Barrier Properties in Endothelial Cells. Nature 325: 253–257.

    Article  CAS  Google Scholar 

  • Jie, H., L. Lian, L.J. Qu, J.X. Zheng, and Z.C. Hou. 2013. Differential Expression of Toll-Like Receptor Genes in Lymphoid Tissues Between Marek’s Disease Virus-Infected and Non Infected Chickens In Chickens. Poultry Science 92: 645–654.

    Article  CAS  Google Scholar 

  • Lin, R.J., B.L. Chang, H.P. Yu, C.L. Liao, and Y.L. Lin. 2006. Blocking of Interferoninduced Jak-Stat Signaling by Japanese Encephalitis Virus NS5 Through a Protein Tyrosine Phosphatase-Mediated Mechanism. Journal of Virology 80: 5908–5918.

    Article  CAS  Google Scholar 

  • Lin, R.J., C.L. Liao, E. Lin, and Y.L. Lin. 2004. Blocking of the Alpha Interferoninduced Jak-Stat Signaling Pathway by Japanese Encephalitis Virus Infection. Journal of Virology 78: 9285–9294.

    Article  CAS  Google Scholar 

  • Nazmi, A., S. Mukherjee, K. Kundu, K. Dutta, A. Mahadevan, S.K. Shankar, and A. Basu. 2014. TLR7 is a Key Regulator of Innate Immunity Against Japanese Encephalitis Virus Infection. Neurobiology of Disease 69: 235–247.

    Article  CAS  Google Scholar 

  • Nowak-Sliwinska, P., T. Segura, and M.L. Iruela-Arispe. 2014. The Chicken Chorioallantoic Membrane Model in Biology, Medicine and Bioengineering. Angiogen 17(4): 779–804.

    Article  CAS  Google Scholar 

  • Olson, M.E., D. Vizzutti, D.W. Morck, and A.K. Cox. 1994. The Parasympatholytic Effects of Atropine Sulfate and Glycopyrrolate in Rats and Rabbits. Canadian Journal of Veterinary Research 58(4): 254–258.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Papahadjopoulos, D., K. Jacobson, G. Poste, and G. Shepherd. 1975. Effects of Local Anesthetics on Membrane Properties. I. Changes in the Fluidity of Phospholipids Bilayers. Biochimica et Biophysica Acta 394: 504–519.

    Article  CAS  Google Scholar 

  • Pieper, J., U. Methner, and A. Berndt. 2008. Heterogeneity of Avian γδ T Cells. Veterinary Immunology and Immunopathology 124: 241–252.

    Article  CAS  Google Scholar 

  • Reed, L.J., and H.A. Muench. 1938. A Simple Method of Estimating Fifty Percent End Points. Journal of Tropical Medicine and Hygiene 27: 493–497.

    Google Scholar 

  • Rothschild, A.M., and E.L.T. Gomes. 1988. Atropine and Hexamethonium-Sensitive, Ca/K-Modulated, Reversible Swelling of Mast Cells In Rat Mesentery, Due to Feeding or Exposure to Carbachol. Agents and Actions 25: 4–10.

    Article  CAS  Google Scholar 

  • Saxena, S.K., P.T. Agrawal, and M.P.N. Nair. 2014. Current Scenario of Antiviral Drugs for Japanese Encephalitis. Journal of Medical Microbiology & Diagnosis 3: 133.

    Google Scholar 

  • Wang, S., Y. Liu, J. Guo, P. Wang, L. Zhang, G. Xiao, and W. Wang. 2017a. Screening of FDA Approved Drugs for Inhibitors of Japanese Encephalitis Virus Infection. Journal of Virology 91: e01055-17.

    Article  Google Scholar 

  • Wang, T., T. Town, L. Alexopoulou, J.F. Anderson, E. Fikrig, et al. 2004. Toll-Like Receptor 3 Mediates West Nile Virus Entry into the Brain Causing Lethal Encephalitis. Nature Medicine 10: 1366–1373.

    Article  CAS  Google Scholar 

  • Wang, Y., Y. Yin, X. Lan, F. Ye, K. Tian, X. Zhao, Y. Huadong, L. Diyan, X. Hengyong, L. Yiping, and Z. Qing. 2017b. Molecular Characterization, Expression of Chicken TBK1 Gene and Its Effect on IRF3 Signaling Pathway. PLoS ONE 12(5): e0177608.

    Article  Google Scholar 

  • Wen, Q., T.J. Fan, and C.L. Tian. 2016. Cytotoxicity of Atropine to Human Corneal Endothelial Cells by Inducing Mitochondrion-Dependent Apoptosis. Experimental Biology and Medicine 241: 1457–1465.

    Article  CAS  Google Scholar 

  • Ye, J., Z. Chen, Y. Li, Z. Zhao, W. He, A. Zohaib, Y. Song, C. Deng, B. Zhang, H. Chen, and S. Cao. 2017. Japanese Encephalitis Virus NS5 Inhibits Type I Interferon (IFN) Production by Blocking the Nuclear Translocation of IFN Regulatory Factor 3 and NF-κB. Journal of Virology 91(8): e00039-17.

    Article  Google Scholar 

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Acknowledgements

This work was supported by CCRH, Ministry of AYUSH, Govt. of India. We also thank the learned reviewers for their valuable suggestions and comments on the earlier version of this manuscript.

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Authors and Affiliations

  1. Department of Virology, Dr. Anjali Chatterjee Regional Research Institute for Homoeopathy, Ministry of AYUSH, Govt. of India, Kolkata, 700035, India

    Urmita Chakraborty, Moonmoon Sinha, Arghyadeep Bhattacharjee, Debabrata Sarkar & Satadal Das

  2. Central Council for Research in Homoeopathy, Ministry of AYUSH, Govt. of India, New Delhi, 110058, India

    Debadatta Nayak, Anil Khurana & Raj Kumar Manchanda

Authors
  1. Urmita Chakraborty
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  2. Moonmoon Sinha
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  3. Arghyadeep Bhattacharjee
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  4. Debadatta Nayak
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  5. Anil Khurana
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  6. Raj Kumar Manchanda
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  7. Debabrata Sarkar
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  8. Satadal Das
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Contributions

UC: Experimentation, manuscript preparation; MS: Experimentation; AB: Experimentation; DN: Idea development; AK: Consultancy; RKM: Idea development; DS: Manuscript preparation; SD: Experiment design and supervising.

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Correspondence to Satadal Das.

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Chakraborty, U., Sinha, M., Bhattacharjee, A. et al. Sub-lethal Dose of Atropine Gives Protection from Japanese Encephalitis Virus Infection in Chick Embryo Model. Proc Zool Soc 73, 235–242 (2020). https://doi.org/10.1007/s12595-020-00324-8

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  • DOI: https://doi.org/10.1007/s12595-020-00324-8

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