Skip to main content

Advertisement

SpringerLink
Log in

Calcitriol [1, 25[OH]2 D3] pre- and post-treatment suppresses inflammatory response to influenza A (H1N1) infection in human lung A549 epithelial cells

  • Original Contribution
  • Published:
European Journal of Nutrition Aims and scope Submit manuscript

Abstract

Purpose

Influenza viruses infect airway epithelial cells, causing respiratory distress. Immune defense is maintained by chemokine/cytokine secretions from airway epithelial cells. While moderate inflammatory response protects from ill effects, hyper-inflammatory response promotes the pathogenesis. High circulating levels of vitamin D are known to mitigate effects of infectious diseases, including respiratory infectious diseases. The question whether and how vitamin D treatment pre-/post-viral exposure modulates inflammatory response is not clear. The present study was undertaken to understand autophagy/apoptosis balance and chemokine/cytokine response to influenza A (H1N1) infection by pre- and post-1, 25-dihydroxyvitamin D3 (1,25[OH]2 D3)[calcitriol] treatment of human lung A549 epithelial cells.

Methods

Influenza A (H1N1) virus was propagated in A549 cell line, titrated using hemagglutination assay, and was used to assess effect of calcitriol. After confirming that 100 nM of calcitriol fails to clear virus, A549 cells were either pre-treated (16 h) with 100 nM or post-treated with 30 nM of 1,25[OH]2 D3 of virus inoculation (1 h). Cells after incubation at 37 °C under 5 % CO2 for 48 h were collected and subjected to RNA and protein extraction. Measurements of viability, influenza M protein, and molecular parameters of cell death and inflammatory response were performed.

Results

We report that treatment of these cells with 100/30 nM of 1,25[OH]2 D3 prior to/or post-H1N1 exposure does not affect viral clearance but significantly reduces autophagy and restores increased apoptosis seen on H1N1 infection back to its constitutive level. However, it significantly decreases the levels of H1N1-induced TNF-α (tumor necrosis factor-alpha), IFN-β (interferon-beta), and IFN-stimulated gene-15 (ISG15). 1,25[OH]2 D3 treatment prior to/or post-H1N1 infection significantly down-regulates IL-8 as well as IL-6 RNA levels. These results demonstrate that calcitriol treatment suppresses the H1N1-induced transcription of the chemokines RANTES and IL-8 in epithelial cells.

Conclusion

The findings provide support for the initiation of vitamin D supplementation program to VDD populations in reducing the severity of influenza.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Rao BL, Yeolekar LR, Kadam SS, Pawar MS, Kulkarni PB, More BA, Khude MR (2005) Influenza surveillance in Pune, India, 2003. Southeast Asian J Trop Med Public Health 36:906–909

    CAS  Google Scholar 

  2. Geiss GK, Salvatore M, Tumpey TM, Carter VS, Wang X, Basler CF, Taubenberger JK, Bumgarner RE, Palese P, Katze MG, Garcia-Sastre A (2002) Cellular transcriptional profiling in influenza A virus-infected lung epithelial cells: the role of the nonstructural NS1 protein in the evasion of the host innate defense and its potential contribution to pandemic influenza. Proc Natl Acad Sci USA 99:10736–10741

    Article  CAS  Google Scholar 

  3. Chakrabarti AK, Pawar SD, Cherian SS, Koratkar SS, Jadhav SM, Pal B (2009) Characterization of the Influenza A H5N1 Viruses of the 2008–09 Outbreaks in India Reveals a Third Introduction and Possible Endemicity. PLoS ONE 4(11):e7846

    Article  Google Scholar 

  4. Li Y, Zhou H, Wen Z, Wu S, Huang C, Jia G, Chen H, Jin M (2011) Transcription analysis on response of swine lung to H1N1 swine influenza virus. BMC Genom 12:398

    Article  CAS  Google Scholar 

  5. Novel Swine-Origin Influenza A (H1N1) Virus Investigation Team. Dawood FS, Jain S, Finelli L, Shaw MW, Lindstrom S, Garten RJ, Gubareva LV, Xu X, Bridges CB, Uyeki TM (2009) Emergence of a novel swine-origin influenza A (H1N1) virus in humans. N Engl J Med 360:2605–2615

  6. Kyriakis CS, Olsen CW, Carman S, Brown IH, Brookes SM, Doorsselaere JV, Reeth KV (2010) Serologic cross-reactivity with pandemic (H1N1) 2009 virus in pigs, Europe. Emerg Infect Dis 16(1):96–99

    Article  Google Scholar 

  7. Loving CL, Brockmeier SL, Vincent AL, Palmer MV, Sacco RE, Nicholson TL (2010) Influenza virus co-infection with Bordetella bronchiseptica enhances bacterial colonization and host responses exacerbating pulmonary lesions. Microb Pathog 49:237–245

    Article  CAS  Google Scholar 

  8. Jo SK, Kim HS, Cho SW, Seo SH (2007) Pathogenesis and inflammatory responses of swine H1N2 influenza viruses in pigs. Virus Res 129:64–70

    Article  CAS  Google Scholar 

  9. Ghosh S, May MJ, Kopp EB (1998) NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol 16:225–260

    Article  CAS  Google Scholar 

  10. Lee SM, Gardy JL, Cheung CY, Cheung TKW, Hui KPY, Ip NY, Guan Y, Hancock REW, Peiris JS (2009) Systems-level comparison of host-responses elicited by avian H5N1 and seasonal H1N1 influenza viruses in primary human macrophages. PLoS ONE 4(12):e8072

    Article  Google Scholar 

  11. Uchide N, Toyoda H (2011) Antioxidant therapy as a potential approach to severe influenza-associated complications. Molecules 16:2032–2052

    Article  CAS  Google Scholar 

  12. Hansdottir S, Monick MM, Hinde SL, Lovan N, Look DC, Hunninghake GW (2008) Respiratory epithelial cells convert inactive vitamin D to its active form: potential effects on host defense. J Immunol 181:7090–7099

    CAS  Google Scholar 

  13. Hansdottir S, Monick MM, Lovan N, Powers L, Gerke A, Hunninghake GW (2010) Vitamin D decreases respiratory syncytial virus induction of NF-kappaB-linked chemokines and cytokines in airway epithelium while maintaining the antiviral state. J Immunol 184:965–974

    Article  CAS  Google Scholar 

  14. Grant WB (2008) Variations in vitamin D production could possibly explain the seasonality of childhood respiratory infections in Hawaii. Pediatr Infect Dis J 27:853

    Google Scholar 

  15. Ginde AA, Mansbach JM, Camargo CA (2009) Association between serum 25-hydroxyvitamin D level and upper respiratory tract infection in the Third National Health and Nutrition Examination Survey. Arch Intern Med 169:384–390

    Article  CAS  Google Scholar 

  16. Cannell JJ, Vieth R, Umhau JC, Holick MF, Grant WB (2006) Epidemic influenza and vitamin D. Epidemiol Infect 134:1129–1140

    Article  CAS  Google Scholar 

  17. Ziegler T, Hall H, Sanchez-Fauquier A, Gamble WC, Cox NJ (1995) Type- and subtype-specific detection of influenza viruses in clinical specimens by rapid culture assay. J Clin Microbiol 33:318–321

    CAS  Google Scholar 

  18. Wayse V, Yousafzai A, Mogale K, Filteau S (2004) Association of subclinical vitamin D deficiency with severe acute lower respiratory infection in Indian children under 5 y. Eur J Clin Nutr 58:563–567

    Article  CAS  Google Scholar 

  19. Ichinohe T, Pang IKS, Iwasaki A (2010) Influenza virus activates inflammasomes via its intracellular M2 channel. Nat Immunol 11(5):404–410

    Article  CAS  Google Scholar 

  20. Cunningham-Rundles S, McNeeley DF, Moon A (2005) Mechanisms of nutrient modulation of the immune response. J Allergy Clin Immunol 115:1119–1128

    Article  CAS  Google Scholar 

  21. Allsup SJ, Shenkin A, Gosney MA (2002) Difficulties of recruitment for a randomized controlled trial involving influenza vaccination in healthy older people. Gerontology 48:170–173

    Article  Google Scholar 

  22. Cannell JJ, Zasloff M, Garland CF, Scragg R, Giovannucci E (2008) On the epidemiology of influenza. Virol J 5:29

    Article  Google Scholar 

  23. Hansdottir S, Monick MM, Hinde SL, Lovan N, Look DC, Hunninghake GW (2008) Respiratory epithelial cells convert inactive vitamin D to its active form: potential effects on host defense. J Immunol 181:7090–7099

    CAS  Google Scholar 

  24. Hansdottir S, Monick MM, Lovan N, Powers L, Gerke A, Hunninghake GW (2010) Vitamin D decreases respiratory syncytial virus induction of NF-kappaB-linked chemokines and cytokines in airway epithelium while maintaining the antiviral state. J Immunol 184:965–974

    Article  CAS  Google Scholar 

  25. Rossman JS, Lamb RA (2009) Autophagy, apoptosis, and the influenza virus M2 protein. Cell Host Microbe 6:299–300

    Article  CAS  Google Scholar 

  26. Zhou Z, Jiang X, Liu D, Fan Z, Hu X, Yan J, Wang M, Gao GF (2009) Autophagy is involved in influenza A virus replication. Autophagy 5(3):321–328

    Article  CAS  Google Scholar 

  27. Gannage M, Dormann D, Albrecht R, Dengjel J, Torossi T, Rämer PC, Lee M, Strowig T, Arrey F, Conenello G, Pypaert M, Andersen J, García-Sastre A, Münz C (2009) Matrix protein 2 of influenza A virus blocks autophagosome fusion with lysosomes. Cell Host Microbe 6:367–380

    Article  CAS  Google Scholar 

  28. Korn T, Oukka M, Kuchroo V, Bettelli E (2007) Th17 cells: effector T cells with inflammatory properties. Semin Immunol 19:362–371

    Article  CAS  Google Scholar 

  29. McGill J, Heusel JW, Legge KL (2009) Innate immune control and regulation of influenza virus infections. J Leukoc Biol 86:803–812

    Article  CAS  Google Scholar 

  30. Miossec P, Korn T, Kuchroo VK (2009) Interleukin-17 and Type 17 helper T cells. N Engl J Med 361:888–898

    Article  CAS  Google Scholar 

  31. To KK, Hung IF, Li IW, Lee KL, Koo CK, Yan WW, Liu R, Ho KY, Chu KH, Watt CL, Luk WK, Lai KY, Chow FL, Mok T, Buckley T, Chan JF, Wong SS, Zheng B, Chen H, Lau CC, Tse H, Cheng VC, Chan KH, Yuen KY (2010) Pandemic H1N1 Study Group. Delayed clearance of viral load and marked cytokine activation in severe cases of pandemic H1N1 2009 influenza virus infection. Clin Infect Dis 50:850–859

    Article  Google Scholar 

  32. Hagau N, Slavcovici A, Gonganau DN, Oltean S, Dirzu DS, Brezoszki ES, Maxim M, Ciuce C, Mlesnite M, Gavrus RL, Laslo C, Hagau R, Petrescu M, Studnicska DM (2010) Clinical aspects and cytokine response in severe H1N1 influenza A virus infection. Crit Care 14:203

    Article  Google Scholar 

  33. Phung TT, Sugamata R, Uno K, Aratani Y, Ozato K, Kawachi S, Nguyen LT, Nakayama T, Suzuki K (2011) Key role of RANTES (regulated upon activation normal T-cell expressed and secreted), nonstructural protein1 and myeloperoxidase in cytokine storm induced by influenza virus PR-8(A/H1N1) infection in A549 bronchial epithelial cells. Microbiol Immunol 55:874–884

    Article  CAS  Google Scholar 

  34. Uchide N, Toyoda H (2011) Antioxidant therapy as a potential approach to severe influenza-associated complications. Molecules 16:2032–2052

    Article  CAS  Google Scholar 

  35. Mark NK, Leung CY, Wei XY, Shen XL, Wong RNS, Leung KN (2004) Inhibition of RANTES expression by indirubin in influenza virus infected human bronchial epithelial cells. Biochem Pharmacol 67:167–174

    Article  Google Scholar 

  36. Kunzi MS, Pitha PM (2003) Interferon targeted genes in host defense. Autoimmunity 36:457–461

    Article  CAS  Google Scholar 

  37. Healy KD, Vanhooke JL, Prahl JM, DeLuca HF (2005) Parathyroid hormone decreases renal vitamin D expression in vivo Proc. Natl Acad Sci USA 102:4724–4728

    Article  CAS  Google Scholar 

  38. Sadeghi K, Wessner B, Laggner U, Ploder M, Tamandl D, Friedl J, Zügel U, Steinmeyer A, Pollak A, Roth E, Boltz-Nitulescu G, Spittler A (2006) Vitamin D3 down-regulates monocyte TLR expression and triggers hyporesponsiveness to pathogen-associated molecular patterns. Eur J Immunol 36:361–370

    Article  CAS  Google Scholar 

  39. Reinhardt TA, Horst RL (1990) Parathyroid hormone down-regulates 1,25-dihydroxyvitamin D receptors (VDR) and VDR messenger ribonucleic acid in vitro and blocks homologous up-regulation of VDR in vivo. Endocrinology 127:942–948

    Article  CAS  Google Scholar 

  40. Klaus G, May T, Hügel U, Von EB, Rodriguez J, Fernandez P, Reichrath J, Ritz E, Mehls O (1997) Parathyroid hormone prevents 1,25 (OH)2D3 induced down-regulation of the vitamin D receptor in growth plate chondrocytes in vitro. Kidney Int 52:45–51

    Article  CAS  Google Scholar 

  41. Sabetta JR, DePetrillo P, Cipriani RJ, Smardin J, Burns LA, Landry ML (2010) Serum 25-hydroxyvitamin D and the incidence of acute viral respiratory tract infections in healthy adults. PLoS ONE 5(6):e11088

    Article  Google Scholar 

Download references

Acknowledgments

We wish to acknowledge Dr. R. Deolankar for initiation of study, and Dr. A. C. Mishra and Dr. M. S. Chadha for supply of influenza strain (both from National Institute of Virology, Pune). The study was supported by Life Science Research Board, DRDO, Government of India grant (MMG) No.LSRB/212/2010-2011.

Conflict of interest

None of the authors had any conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Endocrinology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India

    Drirh Khare, Vishwa Mohan, Gaurav Pandey, Sushil Gupta & Madan M. Godbole

  2. Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India

    Nachiket M. Godbole & Tapan N. Dhole

  3. Department of Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India

    Deepak Kumar

  4. National Institute of Virology, Pune, 411021, India

    Shailesh D. Pawar

Authors
  1. Drirh Khare
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nachiket M. Godbole
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shailesh D. Pawar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vishwa Mohan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gaurav Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sushil Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Deepak Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tapan N. Dhole
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Madan M. Godbole
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Madan M. Godbole.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Khare, D., Godbole, N.M., Pawar, S.D. et al. Calcitriol [1, 25[OH]2 D3] pre- and post-treatment suppresses inflammatory response to influenza A (H1N1) infection in human lung A549 epithelial cells. Eur J Nutr 52, 1405–1415 (2013). https://doi.org/10.1007/s00394-012-0449-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00394-012-0449-7

Keywords

Search

Navigation