1887

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) infection poorly induces pro-inflammatory cytokines (IL-1, IL-6 and TNF-α) and type I IFN production during the early phase of infection. Our microarray analysis indicated strong upregulation of the IL1RA gene in type 2 PRRSV -infected monocyte-derived dendritic cells. Interleukin-1 receptor antagonist (IL-1Ra) is an early inhibitory cytokine that suppresses pro-inflammatory cytokines and T-lymphocyte responses. To investigate the induction of IL-1Ra by PRRSV, monocyte-derived dendritic cells were cultured with type 2 PRRSV or other swine viruses. PRRSV increased both IL1RA gene expression and IL-1Ra protein production in the culture. The enhanced production of IL-1Ra was further confirmed in PRRSV-cultured PBMC and PRRSV-exposed pigs by flow cytometry. Myeloid cell population appeared to be the major IL-1Ra producer both in vitro and in vivo. In contrast to the type 2 PRRSV, the highly pathogenic (HP)- PRRSV did not upregulate IL1RA gene expression in vitro. To determine the kinetics of PRRSV-induced IL1RA gene expression in relation to other pro-inflammatory cytokine genes, PRRSV-negative pigs were vaccinated with a commercially available type 2 modified-live PRRS vaccine or intranasally inoculated with HP-PRRSV. In modified-live PRRS vaccine pigs, upregulation of IL1RA, but not IL1B and IFNA, gene expression was observed from 2 days post- vaccination. Consistent with the in vitro findings, upregulation of IL1RA gene expression was not observed in the HP-PRRSV-infected pigs throughout the experiment. This study identified IL-1Ra as an early immunomodulatory mediator that could be involved in the immunopathogenesis of PRRSV infections.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000665
2017-02-06
2019-09-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/98/1/77.html?itemId=/content/journal/jgv/10.1099/jgv.0.000665&mimeType=html&fmt=ahah

References

  1. Iwasaki A, Pillai PS. Innate immunity to influenza virus infection. Nat Rev Immunol 2014;14:315–328 [CrossRef][PubMed]
    [Google Scholar]
  2. Benedict CA. Viruses and the TNF-related cytokines, an evolving battle. Cytokine Growth Factor Rev 2003;14:349–357 [CrossRef][PubMed]
    [Google Scholar]
  3. Dienz O, Rud JG, Eaton SM, Lanthier PA, Burg E et al. Essential role of IL-6 in protection against H1N1 influenza virus by promoting neutrophil survival in the lung. Mucosal Immunol 2012;5:258–266 [CrossRef][PubMed]
    [Google Scholar]
  4. Oleksowicz L, Dutcher JP. A review of the new cytokines: IL-4, IL-6, IL-11, and IL-12. Am J Ther 1994;1:107–115[PubMed][CrossRef]
    [Google Scholar]
  5. Sims JE, Smith DE. The IL-1 family: regulators of immunity. Nat Rev Immunol 2010;10:117–102 [CrossRef][PubMed]
    [Google Scholar]
  6. Theofilopoulos AN, Baccala R, Beutler B, Kono DH. Type I interferons (alpha/beta) in immunity and autoimmunity. Annu Rev Immunol 2005;23:307–335 [CrossRef][PubMed]
    [Google Scholar]
  7. Rossow KD. Porcine reproductive and respiratory syndrome. Vet Pathol 1998;35:1–20 [CrossRef][PubMed]
    [Google Scholar]
  8. Meng XJ, Paul PS, Halbur PG, Lum MA. Phylogenetic analyses of the putative M (ORF 6) and N (ORF 7) genes of porcine reproductive and respiratory syndrome virus (PRRSV): implication for the existence of two genotypes of PRRSV in the U.S.A. and Europe. Arch Virol 1995;140:745–755 [CrossRef][PubMed]
    [Google Scholar]
  9. Butler JE, Lager KM, Golde W, Faaberg KS, Sinkora M et al. Porcine reproductive and respiratory syndrome (PRRS): an immune dysregulatory pandemic. Immunol Res 2014;59:81–108 [CrossRef][PubMed]
    [Google Scholar]
  10. Lunney JK, Fang Y, Ladinig A, Chen N, Li Y et al. Porcine reproductive and respiratory syndrome virus (PRRSV): pathogenesis and interaction with the immune system. Annu Rev Anim Biosci 2016;4:129–154 [CrossRef][PubMed]
    [Google Scholar]
  11. Rossow KD, Collins JE, Goyal SM, Nelson EA, Christopher-Hennings J et al. Pathogenesis of porcine reproductive and respiratory syndrome virus infection in gnotobiotic pigs. Vet Pathol 1995;32:361–373 [CrossRef][PubMed]
    [Google Scholar]
  12. Amadori M, Razzuoli E. Immune control of PRRS: lessons to be learned and possible ways forward. Front Vet Sci 2014;1:2 [CrossRef][PubMed]
    [Google Scholar]
  13. Li H, Yang H. Infection of porcine reproductive and respiratory syndrome virus suppresses the antibody response to classical swine fever virus vaccination. Vet Microbiol 2003;95:295–301 [CrossRef][PubMed]
    [Google Scholar]
  14. Suradhat S, Kesdangsakonwut S, Sada W, Buranapraditkun S, Wongsawang S et al. Negative impact of porcine reproductive and respiratory syndrome virus infection on the efficacy of classical swine fever vaccine. Vaccine 2006;24:2634–2642 [CrossRef][PubMed]
    [Google Scholar]
  15. Zhou Y, Bai J, Li Y, Wang X, Wang X et al. Suppression of immune responses in pigs by nonstructural protein 1 of porcine reproductive and respiratory syndrome virus. Can J Vet Res 2012;76:255–260[PubMed]
    [Google Scholar]
  16. Zhou Z, Ni J, Cao Z, Han X, Xia Y et al. The epidemic status and genetic diversity of 14 highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) isolates from China in 2009. Vet Microbiol 2011;150:257–269 [CrossRef][PubMed]
    [Google Scholar]
  17. Chen XX, Quan R, Guo XK, Gao L, Shi J et al. Up-regulation of pro-inflammatory factors by HP-PRRSV infection in microglia: implications for HP-PRRSV neuropathogenesis. Vet Microbiol 2014;170:48–57 [CrossRef][PubMed]
    [Google Scholar]
  18. Gao L, Wang L, Huang C, Yang L, Guo XK et al. HP-PRRSV is attenuated by de-optimization of codon pair bias in its RNA-dependent RNA polymerase nsp9 gene. Virology 2015;485:135–144 [CrossRef][PubMed]
    [Google Scholar]
  19. Amarilla SP, Gómez-Laguna J, Carrasco L, Rodríguez-Gómez IM, Caridad Y Ocerín JM et al. A comparative study of the local cytokine response in the lungs of pigs experimentally infected with different PRRSV-1 strains: upregulation of IL-1α in highly pathogenic strain induced lesions. Vet Immunol Immunopathol 2015;164:137–147 [CrossRef][PubMed]
    [Google Scholar]
  20. Liu Y, Shi W, Zhou E, Wang S, Hu S et al. Dynamic changes in inflammatory cytokines in pigs infected with highly pathogenic porcine reproductive and respiratory syndrome virus. Clin Vaccine Immunol 2010;17:1439–1445 [CrossRef][PubMed]
    [Google Scholar]
  21. Flores-Mendoza L, Silva-Campa E, Reséndiz M, Osorio FA, Hernández J. Porcine reproductive and respiratory syndrome virus infects mature porcine dendritic cells and up-regulates interleukin-10 production. Clin Vaccine Immunol 2008;15:720–725 [CrossRef][PubMed]
    [Google Scholar]
  22. van Gorp H, van Breedam W, Delputte PL, Nauwynck HJ. Sialoadhesin and CD163 join forces during entry of the porcine reproductive and respiratory syndrome virus. J Gen Virol 2008;89:2943–2953 [CrossRef][PubMed]
    [Google Scholar]
  23. Wang X, Eaton M, Mayer M, Li H, He D et al. Porcine reproductive and respiratory syndrome virus productively infects monocyte-derived dendritic cells and compromises their antigen-presenting ability. Arch Virol 2007;152:289–303 [CrossRef][PubMed]
    [Google Scholar]
  24. Miller LC, Fox JM. Apoptosis and porcine reproductive and respiratory syndrome virus. Vet Immunol Immunopathol 2004;102:131–142 [CrossRef][PubMed]
    [Google Scholar]
  25. Rodríguez-Gómez IM, Gómez-Laguna J, Carrasco L. Impact of PRRSV on activation and viability of antigen presenting cells. World J Virol 2013;2:146–151 [CrossRef][PubMed]
    [Google Scholar]
  26. Calzada-Nova G, Schnitzlein WM, Husmann RJ, Zuckermann FA. North American porcine reproductive and respiratory syndrome viruses inhibit type I interferon production by plasmacytoid dendritic cells. J Virol 2011;85:2703–2713 [CrossRef][PubMed]
    [Google Scholar]
  27. Patel D, Nan Y, Shen M, Ritthipichai K, Zhu X et al. Porcine reproductive and respiratory syndrome virus inhibits type I interferon signaling by blocking STAT1/STAT2 nuclear translocation. J Virol 2010;84:11045–11055 [CrossRef][PubMed]
    [Google Scholar]
  28. van Gucht S, van Reeth K, Pensaert M. Interaction between porcine reproductive-respiratory syndrome virus and bacterial endotoxin in the lungs of pigs: potentiation of cytokine production and respiratory disease. J Clin Microbiol 2003;41:960–966 [CrossRef][PubMed]
    [Google Scholar]
  29. van Reeth K, Nauwynck H. Proinflammatory cytokines and viral respiratory disease in pigs. Vet Res 2000;31:187–213 [CrossRef][PubMed]
    [Google Scholar]
  30. Lunney JK, Fritz ER, Reecy JM, Kuhar D, Prucnal E et al. Interleukin-8, interleukin-1beta, and interferon-gamma levels are linked to PRRS virus clearance. Viral Immunol 2010;23:127–134 [CrossRef][PubMed]
    [Google Scholar]
  31. Beura LK, Sarkar SN, Kwon B, Subramaniam S, Jones C et al. Porcine reproductive and respiratory syndrome virus nonstructural protein 1beta modulates host innate immune response by antagonizing IRF3 activation. J Virol 2010;84:1574–1584 [CrossRef][PubMed]
    [Google Scholar]
  32. Sagong M, Lee C. Porcine reproductive and respiratory syndrome virus nucleocapsid protein modulates interferon-β production by inhibiting IRF3 activation in immortalized porcine alveolar macrophages. Arch Virol 2011;156:2187–2195 [CrossRef][PubMed]
    [Google Scholar]
  33. Wang R, Zhang YJ. Antagonizing interferon-mediated immune response by porcine reproductive and respiratory syndrome virus. Biomed Res Int 2014;2014:1–9 [CrossRef][PubMed]
    [Google Scholar]
  34. Chung HK, Chae C. Expression of interleukin-10 and interleukin-12 in piglets experimentally infected with porcine reproductive and respiratory syndrome virus (PRRSV). J Comp Pathol 2003;129:205–212 [CrossRef][PubMed]
    [Google Scholar]
  35. Silva-Campa E, Mata-Haro V, Mateu E, Hernández J. Porcine reproductive and respiratory syndrome virus induces CD4 CD8 CD25 Foxp3 regulatory T cells (Tregs). Virology 2012;430:73–80 [CrossRef][PubMed]
    [Google Scholar]
  36. Suradhat S, Thanawongnuwech R, Poovorawan Y. Upregulation of IL-10 gene expression in porcine peripheral blood mononuclear cells by porcine reproductive and respiratory syndrome virus. J Gen Virol 2003;84:453–459 [CrossRef][PubMed]
    [Google Scholar]
  37. Wongyanin P, Buranapraditkul S, Yoo D, Thanawongnuwech R, Roth JA et al. Role of porcine reproductive and respiratory syndrome virus nucleocapsid protein in induction of interleukin-10 and regulatory T-lymphocytes (Treg). J Gen Virol 2012;93:1236–1246 [CrossRef][PubMed]
    [Google Scholar]
  38. Gimeno M, Darwich L, Diaz I, de La Torre E, Pujols J et al. Cytokine profiles and phenotype regulation of antigen presenting cells by genotype-I porcine reproductive and respiratory syndrome virus isolates. Vet Res 2011;42:9 [CrossRef][PubMed]
    [Google Scholar]
  39. Silva-Campa E, Cordoba L, Fraile L, Flores-Mendoza L, Montoya M et al. European genotype of porcine reproductive and respiratory syndrome (PRRSV) infects monocyte-derived dendritic cells but does not induce Treg cells. Virology 2010;396:264–271 [CrossRef][PubMed]
    [Google Scholar]
  40. Arend WP. The balance between IL-1 and IL-1Ra in disease. Cytokine Growth Factor Rev 2002;13:323–340 [CrossRef][PubMed]
    [Google Scholar]
  41. Arend WP, Malyak M, Guthridge CJ, Gabay C. Interleukin-1 receptor antagonist: role in biology. Annu Rev Immunol 1998;16:27–55 [CrossRef][PubMed]
    [Google Scholar]
  42. Delputte PL, Costers S, Nauwynck HJ. Analysis of porcine reproductive and respiratory syndrome virus attachment and internalization: distinctive roles for heparan sulphate and sialoadhesin. J Gen Virol 2005;86:1441–1445 [CrossRef][PubMed]
    [Google Scholar]
  43. Marsh CB, Moore SA, Pope HA, Wewers MD. IL-1ra suppresses endotoxin-induced IL-1 beta and TNF-alpha release from mononuclear phagocytes. Am J Physiol 1994;267:L39–45[PubMed]
    [Google Scholar]
  44. Ludigs K, Parfenov V, du Pasquier RA, Guarda G. Type I IFN-mediated regulation of IL-1 production in inflammatory disorders. Cell Mol Life Sci 2012;69:3395–3418 [CrossRef][PubMed]
    [Google Scholar]
  45. Lee SM, Schommer SK, Kleiboeker SB. Porcine reproductive and respiratory syndrome virus field isolates differ in in vitro interferon phenotypes. Vet Immunol Immunopathol 2004;102:217–231 [CrossRef][PubMed]
    [Google Scholar]
  46. Nan Y, Wang R, Shen M, Faaberg KS, Samal SK et al. Induction of type I interferons by a novel porcine reproductive and respiratory syndrome virus isolate. Virology 2012;432:261–270 [CrossRef][PubMed]
    [Google Scholar]
  47. Bensaude E, Turner JL, Wakeley PR, Sweetman DA, Pardieu C et al. Classical swine fever virus induces proinflammatory cytokines and tissue factor expression and inhibits apoptosis and interferon synthesis during the establishment of long-term infection of porcine vascular endothelial cells. J Gen Virol 2004;85:1029–1037 [CrossRef][PubMed]
    [Google Scholar]
  48. Sládková T, Kostolanský F. The role of cytokines in the immune response to influenza A virus infection. Acta Virol 2006;50:151–162[PubMed]
    [Google Scholar]
  49. van Reeth K. Cytokines in the pathogenesis of influenza. Vet Microbiol 2000;74:109–116 [CrossRef][PubMed]
    [Google Scholar]
  50. Charerntantanakul W. Porcine reproductive and respiratory syndrome virus vaccines: immunogenicity, efficacy and safety aspects. World J Virol 2012;1:23–30 [CrossRef][PubMed]
    [Google Scholar]
  51. Leroith T, Hammond S, Todd SM, Ni Y, Cecere T et al. A modified live PRRSV vaccine and the pathogenic parent strain induce regulatory T cells in pigs naturally infected with Mycoplasma hyopneumoniae. Vet Immunol Immunopathol 2011;140:312–316 [CrossRef][PubMed]
    [Google Scholar]
  52. Guo B, Lager KM, Henningson JN, Miller LC, Schlink SN et al. Experimental infection of United States swine with a Chinese highly pathogenic strain of porcine reproductive and respiratory syndrome virus. Virology 2013;435:372–384 [CrossRef][PubMed]
    [Google Scholar]
  53. Zhang L, Liu J, Bai J, Wang X, Li Y et al. Comparative expression of Toll-like receptors and inflammatory cytokines in pigs infected with different virulent porcine reproductive and respiratory syndrome virus isolates. Virol J 2013;10:135 [CrossRef][PubMed]
    [Google Scholar]
  54. An TQ, Tian ZJ, Xiao Y, Li R, Peng JM et al. Origin of highly pathogenic porcine reproductive and respiratory syndrome virus, China. Emerg Infect Dis 2010;16:365–367 [CrossRef][PubMed]
    [Google Scholar]
  55. Zhou L, Yang H. Porcine reproductive and respiratory syndrome in China. Virus Res 2010;154:31–37 [CrossRef][PubMed]
    [Google Scholar]
  56. Arend WP, Guthridge CJ. Biological role of interleukin 1 receptor antagonist isoforms. Ann Rheum Dis 2000;59 Suppl 1:60i–64 [CrossRef][PubMed]
    [Google Scholar]
  57. Jenkins JK, Arend WP. Interleukin 1 receptor antagonist production in human monocytes is induced by IL-1 alpha, IL-3, IL-4 and GM-CSF. Cytokine 1993;5:407–415 [CrossRef][PubMed]
    [Google Scholar]
  58. Jenkins JK, Malyak M, Arend WP. The effects of interleukin-10 on interleukin-1 receptor antagonist and interleukin-1 beta production in human monocytes and neutrophils. Lymphokine Cytokine Res 1994;13:47–54[PubMed]
    [Google Scholar]
  59. Re F, Mengozzi M, Muzio M, Dinarello CA, Mantovani A et al. Expression of interleukin-1 receptor antagonist (IL-1ra) by human circulating polymorphonuclear cells. Eur J Immunol 1993;23:570–573 [CrossRef][PubMed]
    [Google Scholar]
  60. Song S, Bi J, Wang D, Fang L, Zhang L et al. Porcine reproductive and respiratory syndrome virus infection activates IL-10 production through NF-κB and p38 MAPK pathways in porcine alveolar macrophages. Dev Comp Immunol 2013;39:265–272 [CrossRef][PubMed]
    [Google Scholar]
  61. Banda NK, Guthridge C, Sheppard D, Cairns KS, Muggli M et al. Intracellular IL-1 receptor antagonist type 1 inhibits IL-1-induced cytokine production in keratinocytes through binding to the third component of the COP9 signalosome. J Immunol 2005;174:3608–3616 [CrossRef][PubMed]
    [Google Scholar]
  62. Schiff MH. Role of interleukin 1 and interleukin 1 receptor antagonist in the mediation of rheumatoid arthritis. Ann Rheum Dis 2000;59 Suppl 1:103i–108 [CrossRef][PubMed]
    [Google Scholar]
  63. Mayer-Barber KD, Andrade BB, Oland SD, Amaral EP, Barber DL et al. Host-directed therapy of tuberculosis based on interleukin-1 and type I interferon crosstalk. Nature 2014;511:99–103 [CrossRef][PubMed]
    [Google Scholar]
  64. Matsuki T, Nakae S, Sudo K, Horai R, Iwakura Y. Abnormal T cell activation caused by the imbalance of the IL-1/IL-1R antagonist system is responsible for the development of experimental autoimmune encephalomyelitis. Int Immunol 2006;18:399–407 [CrossRef][PubMed]
    [Google Scholar]
  65. Nakae S, Saijo S, Horai R, Sudo K, Mori S et al. IL-17 production from activated T cells is required for the spontaneous development of destructive arthritis in mice deficient in IL-1 receptor antagonist. Proc Natl Acad Sci U S A 2003;100:5986–5990 [CrossRef][PubMed]
    [Google Scholar]
  66. Wang CC, Fu CL, Yang YH, Lo YC, Wang LC et al. Adenovirus expressing interleukin-1 receptor antagonist alleviates allergic airway inflammation in a murine model of asthma. Gene Ther 2006;13:1414–1421 [CrossRef][PubMed]
    [Google Scholar]
  67. Nakae S, Asano M, Horai R, Iwakura Y. Interleukin-1 beta, but not interleukin-1 alpha, is required for T-cell-dependent antibody production. Immunology 2001;104:402–409 [CrossRef][PubMed]
    [Google Scholar]
  68. Nakae S, Asano M, Horai R, Sakaguchi N, Iwakura Y. IL-1 enhances T cell-dependent antibody production through induction of CD40 ligand and OX40 on T cells. J Immunol 2001;167:90–97 [CrossRef][PubMed]
    [Google Scholar]
  69. Sivaraman V, Pechous RD, Stasulli NM, Eichelberger KR, Miao EA et al. Yersinia pestis activates both IL-1β and IL-1 receptor antagonist to modulate lung inflammation during pneumonic plague. PLoS Pathog 2015;11:e1004688 [CrossRef][PubMed]
    [Google Scholar]
  70. Gresnigt MS, Bozza S, Becker KL, Joosten LA, Abdollahi-Roodsaz S et al. A polysaccharide virulence factor from Aspergillus fumigatus elicits anti-inflammatory effects through induction of Interleukin-1 receptor antagonist. PLoS Pathog 2014;10:e1003936 [CrossRef][PubMed]
    [Google Scholar]
  71. Irikura VM, Hirsch E, Hirsh D. Effects of interleukin-1 receptor antagonist overexpression on infection by Listeria monocytogenes. Infect Immun 1999;67:1901–1909[PubMed]
    [Google Scholar]
  72. Chevalier MF, Petitjean G, Dunyach-Rémy C, Didier C, Girard PM et al. The Th17/Treg ratio, IL-1RA and sCD14 levels in primary HIV infection predict the T-cell activation set point in the absence of systemic microbial translocation. PLoS Pathog 2013;9:e1003453 [CrossRef][PubMed]
    [Google Scholar]
  73. Do TD, Park C, Choi K, Jeong J, Nguyen TT et al. Comparison of experimental infection with northern and southern Vietnamese strains of highly pathogenic porcine reproductive and respiratory syndrome virus. J Comp Pathol 2015;152:227–237 [CrossRef][PubMed]
    [Google Scholar]
  74. Guo B, Lager KM, Schlink SN, Kehrli ME, Brockmeier SL et al. Chinese and Vietnamese strains of HP-PRRSV cause different pathogenic outcomes in United States high health swine. Virology 2013;446:238–250 [CrossRef][PubMed]
    [Google Scholar]
  75. Hill-Batorski L, Halfmann P, Marzi A, Lopes TJ, Neumann G et al. Loss of interleukin 1 receptor antagonist enhances susceptibility to ebola virus infection. J Infect Dis 2015;212:S329–S335 [CrossRef][PubMed]
    [Google Scholar]
  76. Rodríguez-Gómez IM, Käser T, Gómez-Laguna J, Lamp B, Sinn L et al. PRRSV-infected monocyte-derived dendritic cells express high levels of SLA-DR and CD80/86 but do not stimulate PRRSV-naïve regulatory T cells to proliferate. Vet Res 2015;46:54 [CrossRef][PubMed]
    [Google Scholar]
  77. Subramaniam S, Sur JH, Kwon B, Pattnaik AK, Osorio FA. A virulent strain of porcine reproductive and respiratory syndrome virus does not up-regulate interleukin-10 levels in vitro or in vivo. Virus Res 2011;155:415–422 [CrossRef][PubMed]
    [Google Scholar]
  78. Thanawongnuwech R, Amonsin A, Tatsanakit A, Damrongwatanapokin S. Genetics and geographical variation of porcine reproductive and respiratory syndrome virus (PRRSV) in Thailand. Vet Microbiol 2004;101:9–21 [CrossRef][PubMed]
    [Google Scholar]
  79. Ayudhya SN, Assavacheep P, Thanawongnuwech R. One world – one health: the threat of emerging swine diseases. An Asian perspective. Transbound Emerg Dis 2012;59 Suppl 1:9–17 [CrossRef][PubMed]
    [Google Scholar]
  80. Egli C, Thür B, Liu L, Hofmann MA. Quantitative TaqMan RT-PCR for the detection and differentiation of European and North American strains of porcine reproductive and respiratory syndrome virus. J Virol Methods 2001;98:63–75 [CrossRef][PubMed]
    [Google Scholar]
  81. Wongyanin P, Buranapraditkun S, Chokeshai-Usaha K, Thanawonguwech R, Suradhat S. Induction of inducible CD4 CD25 Foxp3 regulatory T lymphocytes by porcine reproductive and respiratory syndrome virus (PRRSV). Vet Immunol Immunopathol 2010;133:170–182 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000665
Loading
/content/journal/jgv/10.1099/jgv.0.000665
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error