Polymorphisms at Residue 222 of the Hemagglutinin of Pandemic Influenza A(H1N1)pdm09: Association of Quasi-Species to Morbidity and Mortality in Different Risk Categories
The D222G substitution in the hemagglutinin (HA) gene of the pandemic influenza A(H1N1)pdm09 virus has been identified as a potential virulence marker, because this change allows for virus invasion deeper into the respiratory tract. In this study, we analyzed D, G and N polymorphisms at residue 222 by pyrosequencing (PSQ). We initially analyzed 401 samples from Brazilian patients. These were categorized with respect to clinical conditions due to influenza infection (mild, serious or fatal) and sub-stratified by risky factors. The frequency of mixed population of virus, with more than one polymorphism at residue 222, was significantly higher in serious (10.6%) and fatal (46.7%) influenza cases, whereas those who showed mild influenza infections were all infected by D222 wild-type. Mixtures of quasi-species showed a significant association of mortality, especially for those with risk factors, in special pregnant women. These results not only reinforce the association between D222G substitution and influenza A(H1N1)pdm09-associated morbidity and mortality, but also add the perspective that a worse clinical prognosis is most likely correlated with mixtures of quasi-species at this HA residue. Therefore, quasi-species may have a critical and underestimated role in influenza-related clinical outcomes.
References
[1]
World Health Organization (2010) Pandemic (H1N1) 2009 - update 112. Geneva. Available: http://www.who.int/csr/don/2010_08_06/en？/index.html.
[2]
Cheng VC, To KK, Tse H, Hung IF, Yuen KY (2012) Two years after pandemic influenza A/2009/H1N1: what have we learned? Clin Microbiol Rev 25: 223–263. doi: 10.1128/cmr.05012-11
[3]
World Health Organization (2010) Preliminary review of D222G amino acid substitution in the haemagglutinin of pandemic influenza A (H1N1) 2009 viruses. Wkly Epidemiol Rec 85: 21–28.
[4]
Chutinimitkul S, Herfst S, Steel J, Lowen AC, Ye J, et al. (2010) Virulence-associated substitution D222G in the hemagglutinin of 2009 pandemic influenza A(H1N1) virus affects receptor binding. J Virol 84: 11802–11813. doi: 10.1128/jvi.01136-10
[5]
Liu Y, Childs RA, Matrosovich T, Wharton S, Palma AS, et al. (2010) Altered receptor specificity and cell tropism of D222G hemagglutinin mutants isolated from fatal cases of pandemic A(H1N1) 2009 influenza virus. J Virol 84: 12069–12074. doi: 10.1128/jvi.01639-10
[6]
Belser JA, Jayaraman A, Raman R, Pappas C, Zeng H, et al. (2011) Effect of D222G Mutation in the Hemagglutinin Protein on Receptor Binding, Pathogenesis and Transmissibility of the 2009 Pandemic H1N1 Influenza Virus. PLoS One 6: e25091. doi: 10.1371/journal.pone.0025091
[7]
Lauring AS, Andino R (2010) Quasispecies theory and the behavior of RNA viruses. PLoS Pathog 6: e1001005. doi: 10.1371/journal.ppat.1001005
[8]
WHO Global Influenza Network (2011) Manual for the laboratory diagnosis and virological surveillance of influenza. pp. 153.
[9]
Souza TM, Salluh JI, Bozza FA, Mesquita M, Soares M, et al. (2010) H1N1pdm influenza infection in hospitalized cancer patients: clinical evolution and viral analysis. PLoS One 5: e14158. doi: 10.1371/journal.pone.0014158
[10]
Levine M, Sheu TG, Gubareva LV, Mishin VP (2011) Detection of hemagglutinin variants of the pandemic influenza A (H1N1) 2009 virus by pyrosequencing. J Clin Microbiol 49: 1307–1312. doi: 10.1128/jcm.02424-10
[11]
Vazquez-Perez JA, Isa P, Kobasa D, Ormsby CE, Ramirez-Gonzalez JE, et al. (2013) A (H1N1) pdm09 HA D222 variants associated with severity and mortality in patients during a second wave in Mexico. Virol J 10: 41. doi: 10.1186/1743-422x-10-41
[12]
Yasugi M, Nakamura S, Daidoji T, Kawashita N, Ramadhany R, et al. (2012) Frequency of D222G and Q223R hemagglutinin mutants of pandemic (H1N1) 2009 influenza virus in Japan between 2009 and 2010. PLoS One 7: e30946. doi: 10.1371/journal.pone.0030946
[13]
Ministério da Saúde (2010) Influenza Pandêmica (H1N1) 2009 – Análise da situa？？o epidemiológica e da resposta no ano de 2009. In: Saúde Sdve, editor. Brasilia. pp. 21.
[14]
Wedde M, Wahlisch S, Wolff T, Schweiger B (2013) Predominance of HA-222D/G Polymorphism in Influenza A(H1N1)pdm09 Viruses Associated with Fatal and Severe Outcomes Recently Circulating in Germany. PLoS One 8: e57059. doi: 10.1371/journal.pone.0057059
[15]
Selleri M, Piralla A, Rozera G, Giombini E, Bartolini B, et al.. (2012) Detection of haemagglutinin D222 polymorphisms in influenza A(H1N1)pdm09-infected patients by ultra-deep pyrosequencing. Clin Microbiol Infect.
[16]
Wang B, Dwyer DE, Soedjono M, Shi H, Matlho K, et al.. (2011) Evidence of the circulation of pandemic influenza (H1N1) 2009 with D222D/G/N/S hemagglutinin polymorphisms during the first wave of the 2009 influenza pandemic. J Clin Virol.
[17]
Kuroda M, Katano H, Nakajima N, Tobiume M, Ainai A, et al. (2010) Characterization of quasispecies of pandemic 2009 influenza A virus (A/H1N1/2009) by de novo sequencing using a next-generation DNA sequencer. PLoS One 5: e10256. doi: 10.1371/journal.pone.0010256
[18]
Chen H, Wen X, To KK, Wang P, Tse H, et al. (2010) Quasispecies of the D225G substitution in the hemagglutinin of pandemic influenza A(H1N1) 2009 virus from patients with severe disease in Hong Kong, China. J Infect Dis 201: 1517–1521. doi: 10.1086/652661
[19]
Rykkvin R, Kilander A, Dudman S, Hungnes O (2013) Within-patient emergence of the influenza A(H1N1)pdm09 HA1 222G variant and clear association with severe disease, Norway. Euro Surveill 18.
[20]
Puzelli S, Facchini M, Spagnolo D, De Marco MA, Calzoletti L, et al. (2010) Transmission of hemagglutinin D222G mutant strain of pandemic (H1N1) 2009 virus. Emerg Infect Dis 16: 863–865. doi: 10.3201/eid1605.091858
[21]
Houng HS, Garner J, Zhou Y, Lyons A, Kuschner R, et al. (2012) Emergent 2009 influenza A(H1N1) viruses containing HA D222N mutation associated with severe clinical outcomes in the Americas. J Clin Virol 53: 12–15. doi: 10.1016/j.jcv.2011.09.004
[22]
Baldanti F, Campanini G, Piralla A, Rovida F, Braschi A, et al. (2010) Severe outcome of influenza A/H1N1/09v infection associated with 222G/N polymorphisms in the haemagglutinin: a multicentre study. Clin Microbiol Infect 17: 1166–1169. doi: 10.1111/j.1469-0691.2010.03403.x
[23]
Kilander A, Rykkvin R, Dudman SG, Hungnes O (2010) Observed association between the HA1 mutation D222G in the 2009 pandemic influenza A(H1N1) virus and severe clinical outcome, Norway 2009-2010. Euro Surveill 15.
[24]
Ledesma J, Pozo F, Ruiz MP, Navarro JM, Pineiro L, et al. (2011) Substitutions in position 222 of haemagglutinin of pandemic influenza A (H1N1) 2009 viruses in Spain. J Clin Virol 51: 75–78. doi: 10.1016/j.jcv.2011.01.020
[25]
Mak GC, Au KW, Tai LS, Chuang KC, Cheng KC, et al.. (2010) Association of D222G substitution in haemagglutinin of 2009 pandemic influenza A (H1N1) with severe disease. Euro Surveill 15.
[26]
Miller RR, MacLean AR, Gunson RN, Carman WF (2010) Occurrence of haemagglutinin mutation D222G in pandemic influenza A(H1N1) infected patients in the West of Scotland, United Kingdom, 2009–10. Euro Surveill 15.
[27]
Mosby LG, Rasmussen SA, Jamieson DJ (2011) 2009 pandemic influenza A (H1N1) in pregnancy: a systematic review of the literature. Am J Obstet Gynecol 205: 10–18. doi: 10.1016/j.ajog.2010.12.033
[28]
Orczyk-Pawilowicz M, Augustyniak D, Hirnle L, Katnik-Prastowska I (2012) Lectin-based analysis of fucose and sialic acid expressions on human amniotic IgA during normal pregnancy. Glycoconj J.
[29]
Chan KH, Zhang AJ, To KK, Chan CC, Poon VK, et al. (2010) Wild type and mutant 2009 pandemic influenza A (H1N1) viruses cause more severe disease and higher mortality in pregnant BALB/c mice. PLoS One 5: e13757. doi: 10.1371/journal.pone.0013757
