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Eosinophils pp 257–266Cite as

Eosinophils and Respiratory Virus Infection: A Dual-Standard Curve qRT-PCR-Based Method for Determining Virus Recovery from Mouse Lung Tissue

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1178))

Abstract

Several lines of investigation have indicated a role for eosinophilic leukocytes in limiting virus infectivity and promoting virion clearance. We have established a respiratory virus infection model with pneumonia virus of mice (PVM; family Paramyxoviridae), a natural mouse pathogen that replicates the more severe forms of human disease elicited by the phylogenetically related respiratory syncytial virus (RSV). In this chapter, we present a rapid and highly reproducible dual-standard curve qRT-PCR based method for quantitative detection of PVM replication in mouse lung tissue. We have used this assay to evaluate eosinophil-mediated antiviral host defense in mouse models of cytokine and antigen-driven eosinophilic inflammation.

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References

  1. Edwards MR, Bartlett NW, Hussell T, Openshaw P, Johnston SL (2012) The microbiology of asthma. Nat Rev Microbiol 10: 459–471

    CAS  PubMed  Google Scholar 

  2. Gibson PG (2009) Inflammatory phenotypes in adult asthma: clinical applications. Clin Respir J 3:198–206

    Article  PubMed  Google Scholar 

  3. Lin TY, Poon AH, Hamid Q (2013) Asthma phenotypes and endotypes. Curr Opin Pulm Med 19:18–23

    Article  CAS  PubMed  Google Scholar 

  4. Walsh GM (2013) Profile of reslizumab in eosinophilic disease and its potential in the treatment of poorly controlled eosinophilic asthma. Biologics 7:7–11

    CAS  PubMed Central  PubMed  Google Scholar 

  5. Castro M, Mathur S, Hargreave F, Boulet LP, Xie F, Young J, Wilkins HJ, Henkel T, Nair P, Res-5-0010 Study Group (2010) Reslizumab for poorly controlled, eosinophilic asthma: a randomized, placebo controlled study. Am J Respir Crit Care Med 184:1125–1132

    Article  Google Scholar 

  6. Thomas A, Busse W (2012) The evolving role of eosinophils in asthma. In: Lee JJ, Rosenberg HF (eds) Eosinophils in health and disease. Elsevier, Inc., Waltham, MA, pp 448–461

    Google Scholar 

  7. Rosenberg HF, Dyer KD, Foster PS (2013) Eosinophils: changing perspectives in health and disease. Nat Rev Immunol 13:9–22

    Article  CAS  PubMed  Google Scholar 

  8. Domachowske JB, Dyer KD, Bonville CA, Rosenberg HF (1998) Recombinant human eosinophil-derived neurotoxin/RNase 2 functions as an effective antiviral agent against respiratory syncytial virus. J Infect Dis 177: 1458–1464

    Article  CAS  PubMed  Google Scholar 

  9. Adamko DJ, Yost BL, Gleich GJ, Fryer AD, Jacoby DB (1999) Ovalbumin sensitization changes the inflammatory response to subsequent parainfluenza infection. Eosinophils mediate airway hyperresponsiveness, m(2) muscarinic receptor dysfunction and antiviral effects. J Exp Med 190:1465–1478

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. Phipps S, Lam CE, Mahalingam S, Newhouse M, Ramirez R, Rosenberg HF, Foster PS, Matthaei KI (2007) Eosinophils contribute to innate antiviral immunity and promote clearance of respiratory syncytial virus. Blood 110:1578–1586

    Article  CAS  PubMed  Google Scholar 

  11. Percopo CM, Dyer KD, Ochkur SI, Luo JL, Fischer ER, Lee JJ, Lee NA, Domachowske JB, Rosenberg HF (2014) Activated mouse eosinophils protect against lethal respiratory virus infection. Blood. 123:743–752

    Google Scholar 

  12. Bem RA, Domachowske JB, Rosenberg HF (2011) Animal models of human respiratory syncytial virus disease. Am J Physiol Lung Cell Mol Physiol 301:L148–L156

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Gabryszewski SJ, Bachar O, Dyer KD, Percopo CM, Killoran KE, Domachowske JB, Rosenberg HF (2011) Lactobacilllus-mediated priming of the respiratory mucosa protects against lethal pneumovirus infection. J Immunol 186:1151–1161

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Garcia-Crespo KE, Chan CC, Gabryszewski SJ, Percopo CM, Rigaux P, Dyer KD, Domachowske JB, Rosenberg HF (2013) Lactobacillus priming of the respiratory tract: heterologous immunity and protection against lethal pneumovirus infection. Antiviral Res 97:270–279

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Glineur SF, Renshaw RW, Percopo CM, Dyer KD, Dubovi EJ, Domachowske JB, Rosenberg HF (2013) Novel pneumoviruses (PnVs): evolution and inflammatory pathology. Virology 443:257–264

    Article  CAS  PubMed  Google Scholar 

  16. Ochkur SI, Jacobsen EA, Protheroe CA, Biechele TL, Pero RS, McGarry MP, Wang H, O’Neill KR, Colbert DC, Colby TV, Shen H, Blackburn MR, Irvin CC, Lee JJ, Lee NA (2007) Co-expression of IL-5 and eotaxin-2 in mice creates an eosinophil-dependent model of respiratory inflammation with characteristics of severe asthma. J Immunol 178: 7879–7889

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

Support provided by NIAID Division of Intramural Research (AI000941 and AI000943) to HFR and Children’s Miracle Network of New York to JBD.

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Authors and Affiliations

  1. Inflammation Immunobiology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9000 Rockville Pike, 20892, Bethesda, MD, USA

    Caroline M. Percopo M.S., Kimberly D. Dyer Ph.D. & Kendal A. Karpe B.S.

  2. Division of Infectious Diseases, Department of Pediatrics, SUNY Upstate Medical University, 750 East Adams Street, 13210, Syracuse, NY, USA

    Joseph B. Domachowske M.D.

  3. Laboratory of Allergic Diseases, Inflammation Immunobiology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 10, Room 11C215, 9000 Rockville Pike, 20892, Bethesda, MD, USA

    Helene F. Rosenberg M.D., Ph.D.

Authors
  1. Caroline M. Percopo M.S.
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  2. Kimberly D. Dyer Ph.D.
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  3. Kendal A. Karpe B.S.
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  4. Joseph B. Domachowske M.D.
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  5. Helene F. Rosenberg M.D., Ph.D.
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Corresponding author

Correspondence to Helene F. Rosenberg M.D., Ph.D. .

Editor information

Editors and Affiliations

  1. Division of Applied Medicine, University of Aberdeen Institute of Medical Sciences, Aberdeen, United Kingdom

    Garry M. Walsh

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Percopo, C.M., Dyer, K.D., Karpe, K.A., Domachowske, J.B., Rosenberg, H.F. (2014). Eosinophils and Respiratory Virus Infection: A Dual-Standard Curve qRT-PCR-Based Method for Determining Virus Recovery from Mouse Lung Tissue. In: Walsh, G. (eds) Eosinophils. Methods in Molecular Biology, vol 1178. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1016-8_22

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  • DOI: https://doi.org/10.1007/978-1-4939-1016-8_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1015-1

  • Online ISBN: 978-1-4939-1016-8

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