HIV-1 variants are archived throughout infection and persist in the reservoir


Autoři: Kelsie Brooks aff001;  Bradley R. Jones aff002;  Dario A. Dilernia aff001;  Daniel J. Wilkins aff001;  Daniel T. Claiborne aff001;  Samantha McInally aff001;  Jill Gilmour aff003;  William Kilembe aff004;  Jeffrey B. Joy aff002;  Susan A. Allen aff004;  Zabrina L. Brumme aff002;  Eric Hunter aff001
Působiště autorů: Emory Vaccine Center, Emory University, Atlanta, Georgia, United States of America aff001;  British Columbia Centre for Excellence in HIV/AIDS, Vancouver, British Columbia, Canada aff002;  Human Immunology Lab, International AIDS Vaccine Initiative, London, England, United Kingdom aff003;  Zambia-Emory HIV Research Project, Lusaka, Zambia aff004;  Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada aff005;  Department of Pathology & Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, United States of America aff006;  Faculty of Health Sciences, Simon Fraser University, Burnaby, British Columbia, Canada aff007
Vyšlo v časopise: HIV-1 variants are archived throughout infection and persist in the reservoir. PLoS Pathog 16(6): e32767. doi:10.1371/journal.ppat.1008378
Kategorie: Research Article
doi: 10.1371/journal.ppat.1008378

Souhrn

The HIV-1 reservoir consists of latently infected cells that persist despite antiretroviral therapy (ART). Elucidating the proviral genetic composition of the reservoir, particularly in the context of pre-therapy viral diversity, is therefore important to understanding reservoir formation and the persistence of latently infected cells. Here we investigate reservoir proviral variants from 13 Zambian acutely-infected individuals with additional pre-therapy sampling for a unique comparison to the ART-naïve quasispecies. We identified complete transmitted/founder (TF) viruses from seroconversion plasma samples, and additionally amplified and sequenced HIV-1 from plasma obtained one year post-infection and just prior to ART initiation. While the majority of proviral variants in the reservoir were most closely related to viral variants from the latest pre-therapy time point, we also identified reservoir proviral variants dating to or near the time of infection, and to intermediate time points between infection and treatment initiation. Reservoir proviral variants differing by five or fewer nucleotide changes from the TF virus persisted during treatment in five individuals, including proviral variants that exactly matched the TF in two individuals, one of whom had remained ART-naïve for more than six years. Proviral variants during treatment were significantly less divergent from the TF virus than plasma variants present at the last ART-naïve time point. These findings indicate that reservoir proviral variants are archived throughout infection, recapitulating much of the viral diversity that arises throughout untreated HIV-1 infection, and strategies to target and reduce the reservoir must therefore permit for the clearance of proviruses encompassing this extensive diversity.

Klíčová slova:

Antiretroviral therapy – HIV – HIV-1 – Phylogenetic analysis – Phylogenetics – Polymerase chain reaction – Sequence analysis – Viral replication


Zdroje

1. UNAIDS. Fact sheet: global AIDS update 2019. 2019.

2. Hutter G, Nowak D, Mossner M, Ganepola S, Mussig A, Allers K, et al. Long-term control of HIV by CCR5 Delta32/Delta32 stem-cell transplantation. N Engl J Med. 2009;360(7):692–8. Epub 2009/02/14. doi: 10.1056/NEJMoa0802905 19213682.

3. Allers K, Hutter G, Hofmann J, Loddenkemper C, Rieger K, Thiel E, et al. Evidence for the cure of HIV infection by CCR5Delta32/Delta32 stem cell transplantation. Blood. 2011;117(10):2791–9. Epub 2010/12/15. doi: 10.1182/blood-2010-09-309591 21148083.

4. Gupta RK, Abdul-Jawad S, McCoy LE, Mok HP, Peppa D, Salgado M, et al. HIV-1 remission following CCR5Delta32/Delta32 haematopoietic stem-cell transplantation. Nature. 2019;568(7751):244–8. Epub 2019/03/06. doi: 10.1038/s41586-019-1027-4 30836379.

5. Finzi D, Hermankova M, Pierson T, Carruth LM, Buck C, Chaisson RE, et al. Identification of a reservoir for HIV-1 in patients on highly active antiretroviral therapy. Science (New York, NY). 1997;278(5341):1295–300. Epub 1997/11/21. doi: 10.1126/science.278.5341.1295 9360927.

6. Wong JK, Hezareh M, Gunthard HF, Havlir DV, Ignacio CC, Spina CA, et al. Recovery of replication-competent HIV despite prolonged suppression of plasma viremia. Science (New York, NY). 1997;278(5341):1291–5. Epub 1997/11/21. doi: 10.1126/science.278.5341.1291 9360926.

7. Chun TW, Stuyver L, Mizell SB, Ehler LA, Mican JA, Baseler M, et al. Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. Proceedings of the National Academy of Sciences of the United States of America. 1997;94(24):13193–7. Epub 1997/12/16. doi: 10.1073/pnas.94.24.13193 9371822; PubMed Central PMCID: PMC24285.

8. Chomont N, El-Far M, Ancuta P, Trautmann L, Procopio FA, Yassine-Diab B, et al. HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. Nature medicine. 2009;15(8):893–900. Epub 2009/06/23. doi: 10.1038/nm.1972 19543283; PubMed Central PMCID: PMC2859814.

9. Maldarelli F, Wu X, Su L, Simonetti FR, Shao W, Hill S, et al. HIV latency. Specific HIV integration sites are linked to clonal expansion and persistence of infected cells. Science (New York, NY). 2014;345(6193):179–83. Epub 2014/06/28. doi: 10.1126/science.1254194 24968937; PubMed Central PMCID: PMC4262401.

10. Wagner TA, McLaughlin S, Garg K, Cheung CY, Larsen BB, Styrchak S, et al. HIV latency. Proliferation of cells with HIV integrated into cancer genes contributes to persistent infection. Science (New York, NY). 2014;345(6196):570–3. Epub 2014/07/12. doi: 10.1126/science.1256304 25011556; PubMed Central PMCID: PMC4230336.

11. Wang Z, Gurule EE, Brennan TP, Gerold JM, Kwon KJ, Hosmane NN, et al. Expanded cellular clones carrying replication-competent HIV-1 persist, wax, and wane. Proceedings of the National Academy of Sciences of the United States of America. 2018;115(11):E2575–E84. Epub 2018/02/28. doi: 10.1073/pnas.1720665115 29483265; PubMed Central PMCID: PMC5856552.

12. Finzi D, Blankson J, Siliciano JD, Margolick JB, Chadwick K, Pierson T, et al. Latent infection of CD4+ T cells provides a mechanism for lifelong persistence of HIV-1, even in patients on effective combination therapy. Nature medicine. 1999;5(5):512–7. Epub 1999/05/06. doi: 10.1038/8394 10229227.

13. Siliciano JD, Kajdas J, Finzi D, Quinn TC, Chadwick K, Margolick JB, et al. Long-term follow-up studies confirm the stability of the latent reservoir for HIV-1 in resting CD4+ T cells. Nature medicine. 2003;9(6):727–8. Epub 2003/05/20. doi: 10.1038/nm880 12754504.

14. Zhang L, Chung C, Hu BS, He T, Guo Y, Kim AJ, et al. Genetic characterization of rebounding HIV-1 after cessation of highly active antiretroviral therapy. J Clin Invest. 2000;106(7):839–45. Epub 2000/10/06. doi: 10.1172/JCI10565 11018071; PubMed Central PMCID: PMC517816.

15. Joos B, Fischer M, Kuster H, Pillai SK, Wong JK, Boni J, et al. HIV rebounds from latently infected cells, rather than from continuing low-level replication. Proceedings of the National Academy of Sciences of the United States of America. 2008;105(43):16725–30. Epub 2008/10/22. doi: 10.1073/pnas.0804192105 18936487; PubMed Central PMCID: PMC2575487.

16. Kearney MF, Spindler J, Shao W, Yu S, Anderson EM, O'Shea A, et al. Lack of detectable HIV-1 molecular evolution during suppressive antiretroviral therapy. PLoS pathogens. 2014;10(3):e1004010. Epub 2014/03/22. doi: 10.1371/journal.ppat.1004010 24651464; PubMed Central PMCID: PMC3961343.

17. Kearney MF, Wiegand A, Shao W, Coffin JM, Mellors JW, Lederman M, et al. Origin of Rebound Plasma HIV Includes Cells with Identical Proviruses That Are Transcriptionally Active before Stopping of Antiretroviral Therapy. Journal of virology. 2016;90(3):1369–76. Epub 2015/11/20. doi: 10.1128/JVI.02139-15 26581989; PubMed Central PMCID: PMC4719635.

18. Cohen YZ, Lorenzi JCC, Krassnig L, Barton JP, Burke L, Pai J, et al. Relationship between latent and rebound viruses in a clinical trial of anti-HIV-1 antibody 3BNC117. The Journal of experimental medicine. 2018;215(9):2311–24. Epub 2018/08/04. doi: 10.1084/jem.20180936 30072495; PubMed Central PMCID: PMC6122972.

19. Palich R, Ghosn J, Chaillon A, Boilet V, Nere ML, Chaix ML, et al. Viral rebound in semen after antiretroviral treatment interruption in an HIV therapeutic vaccine double-blind trial. AIDS (London, England). 2019;33(2):279–84. Epub 2018/10/17. doi: 10.1097/QAD.0000000000002058 30325777.

20. Colby DJ, Trautmann L, Pinyakorn S, Leyre L, Pagliuzza A, Kroon E, et al. Rapid HIV RNA rebound after antiretroviral treatment interruption in persons durably suppressed in Fiebig I acute HIV infection. Nature medicine. 2018;24(7):923–6. Epub 2018/06/13. doi: 10.1038/s41591-018-0026-6 29892063; PubMed Central PMCID: PMC6092240.

21. Vibholm LK, Lorenzi JCC, Pai JA, Cohen YZ, Oliveira TY, Barton JP, et al. Characterization of Intact Proviruses in Blood and Lymph Node from HIV-Infected Individuals Undergoing Analytical Treatment Interruption. Journal of virology. 2019;93(8):e01920–18. Epub 2019/02/01. doi: 10.1128/JVI.01920-18 30700598; PubMed Central PMCID: PMC6450127.

22. Ho YC, Shan L, Hosmane NN, Wang J, Laskey SB, Rosenbloom DI, et al. Replication-competent noninduced proviruses in the latent reservoir increase barrier to HIV-1 cure. Cell. 2013;155(3):540–51. Epub 2013/11/19. doi: 10.1016/j.cell.2013.09.020 24243014; PubMed Central PMCID: PMC3896327.

23. Buzon MJ, Sun H, Li C, Shaw A, Seiss K, Ouyang Z, et al. HIV-1 persistence in CD4+ T cells with stem cell-like properties. Nature medicine. 2014;20(2):139–42. Epub 2014/01/15. doi: 10.1038/nm.3445 24412925; PubMed Central PMCID: PMC3959167.

24. Bruner KM, Murray AJ, Pollack RA, Soliman MG, Laskey SB, Capoferri AA, et al. Defective proviruses rapidly accumulate during acute HIV-1 infection. Nature medicine. 2016;22(9):1043–9. Epub 2016/08/09. doi: 10.1038/nm.4156 27500724; PubMed Central PMCID: PMC5014606.

25. Hiener B, Horsburgh BA, Eden JS, Barton K, Schlub TE, Lee E, et al. Identification of Genetically Intact HIV-1 Proviruses in Specific CD4(+) T Cells from Effectively Treated Participants. Cell reports. 2017;21(3):813–22. Epub 2017/10/19. doi: 10.1016/j.celrep.2017.09.081 29045846; PubMed Central PMCID: PMC5960642.

26. Bruner KM, Wang Z, Simonetti FR, Bender AM, Kwon KJ, Sengupta S, et al. A quantitative approach for measuring the reservoir of latent HIV-1 proviruses. Nature. 2019;566(7742):120–5. Epub 2019/02/01. doi: 10.1038/s41586-019-0898-8 30700913; PubMed Central PMCID: PMC6447073.

27. Daar ES, Bai J, Hausner MA, Majchrowicz M, Tamaddon M, Giorgi JV. Acute HIV syndrome after discontinuation of antiretroviral therapy in a patient treated before seroconversion. Ann Intern Med. 1998;128(10):827–9. Epub 1998/05/23. doi: 10.7326/0003-4819-128-10-199805150-00005 9599194.

28. Whitney JB, Hill AL, Sanisetty S, Penaloza-MacMaster P, Liu J, Shetty M, et al. Rapid seeding of the viral reservoir prior to SIV viraemia in rhesus monkeys. Nature. 2014;512(7512):74–7. Epub 2014/07/22. doi: 10.1038/nature13594 25042999; PubMed Central PMCID: PMC4126858.

29. Henrich TJ, Hatano H, Bacon O, Hogan LE, Rutishauser R, Hill A, et al. HIV-1 persistence following extremely early initiation of antiretroviral therapy (ART) during acute HIV-1 infection: An observational study. PLoS Med. 2017;14(11):e1002417. Epub 2017/11/08. doi: 10.1371/journal.pmed.1002417 29112956; PubMed Central PMCID: PMC5675377.

30. Brodin J, Zanini F, Thebo L, Lanz C, Bratt G, Neher RA, et al. Establishment and stability of the latent HIV-1 DNA reservoir. eLife. 2016;5(2050-084X (Electronic)). Epub 2016/11/18. doi: 10.7554/eLife.18889 27855060; PubMed Central PMCID: PMC5201419.

31. Abrahams MR, Joseph SB, Garrett N, Tyers L, Moeser M, Archin N, et al. The replication-competent HIV-1 latent reservoir is primarily established near the time of therapy initiation. Sci Transl Med. 2019;11(513):eaaw5589. Epub 2019/10/11. doi: 10.1126/scitranslmed.aaw5589 31597754.

32. Jones BR, Kinloch NN, Horacsek J, Ganase B, Harris M, Harrigan PR, et al. Phylogenetic approach to recover integration dates of latent HIV sequences within-host. Proceedings of the National Academy of Sciences of the United States of America. 2018;115(38):E8958–E67. Epub 2018/09/07. doi: 10.1073/pnas.1802028115 30185556; PubMed Central PMCID: PMC6156657.

33. Dilernia DA, Chien JT, Monaco DC, Brown MP, Ende Z, Deymier MJ, et al. Multiplexed highly-accurate DNA sequencing of closely-related HIV-1 variants using continuous long reads from single molecule, real-time sequencing. Nucleic acids research. 2015;43(20):e129. Epub 2015/06/24. doi: 10.1093/nar/gkv630 26101252; PubMed Central PMCID: PMC4787755.

34. Theys K, Libin P, Pineda-Pena AC, Nowe A, Vandamme AM, Abecasis AB. The impact of HIV-1 within-host evolution on transmission dynamics. Curr Opin Virol. 2018;28:92–101. Epub 2017/12/25. doi: 10.1016/j.coviro.2017.12.001 29275182.

35. Yerly S, Perneger TV, Vora S, Hirschel B, Perrin L. Decay of cell-associated HIV-1 DNA correlates with residual replication in patients treated during acute HIV-1 infection. AIDS (London, England). 2000;14(18):2805–12. Epub 2001/01/12. doi: 10.1097/00002030-200012220-00001 11153661.

36. Blankson JN, Finzi D, Pierson TC, Sabundayo BP, Chadwick K, Margolick JB, et al. Biphasic decay of latently infected CD4+ T cells in acute human immunodeficiency virus type 1 infection. The Journal of infectious diseases. 2000;182(6):1636–42. Epub 2000/11/09. doi: 10.1086/317615 11069234.

37. Hocqueloux L, Avettand-Fenoel V, Jacquot S, Prazuck T, Legac E, Melard A, et al. Long-term antiretroviral therapy initiated during primary HIV-1 infection is key to achieving both low HIV reservoirs and normal T cell counts. J Antimicrob Chemother. 2013;68(5):1169–78. Epub 2013/01/22. doi: 10.1093/jac/dks533 23335199.

38. Besson GJ, Lalama CM, Bosch RJ, Gandhi RT, Bedison MA, Aga E, et al. HIV-1 DNA decay dynamics in blood during more than a decade of suppressive antiretroviral therapy. Clin Infect Dis. 2014;59(9):1312–21. Epub 2014/07/31. doi: 10.1093/cid/ciu585 25073894; PubMed Central PMCID: PMC4200019.

39. Archin NM, Vaidya NK, Kuruc JD, Liberty AL, Wiegand A, Kearney MF, et al. Immediate antiviral therapy appears to restrict resting CD4+ cell HIV-1 infection without accelerating the decay of latent infection. Proceedings of the National Academy of Sciences of the United States of America. 2012;109(24):9523–8. Epub 2012/05/31. doi: 10.1073/pnas.1120248109 22645358; PubMed Central PMCID: PMC3386138.

40. Buzon MJ, Martin-Gayo E, Pereyra F, Ouyang Z, Sun H, Li JZ, et al. Long-term antiretroviral treatment initiated at primary HIV-1 infection affects the size, composition, and decay kinetics of the reservoir of HIV-1-infected CD4 T cells. Journal of virology. 2014;88(17):10056–65. Epub 2014/06/27. doi: 10.1128/JVI.01046-14 24965451; PubMed Central PMCID: PMC4136362.

41. Ruff CT, Ray SC, Kwon P, Zinn R, Pendleton A, Hutton N, et al. Persistence of wild-type virus and lack of temporal structure in the latent reservoir for human immunodeficiency virus type 1 in pediatric patients with extensive antiretroviral exposure. Journal of virology. 2002;76(18):9481–92. Epub 2002/08/21. doi: 10.1128/jvi.76.18.9481-9492.2002 12186930; PubMed Central PMCID: PMC136462.

42. Verhofstede C, Noe A, Demecheleer E, De Cabooter N, Van Wanzeele F, Van Der Gucht B, et al. Drug-resistant variants that evolve during nonsuppressive therapy persist in HIV-1-infected peripheral blood mononuclear cells after long-term highly active antiretroviral therapy. Journal of acquired immune deficiency syndromes. 2004;35(5):473–83. Epub 2004/03/17. doi: 10.1097/00126334-200404150-00005 15021312.

43. Kieffer TL, Finucane MM, Nettles RE, Quinn TC, Broman KW, Ray SC, et al. Genotypic analysis of HIV-1 drug resistance at the limit of detection: virus production without evolution in treated adults with undetectable HIV loads. The Journal of infectious diseases. 2004;189(8):1452–65. Epub 2004/04/10. doi: 10.1086/382488 15073683.

44. Bailey JR, Sedaghat AR, Kieffer T, Brennan T, Lee PK, Wind-Rotolo M, et al. Residual human immunodeficiency virus type 1 viremia in some patients on antiretroviral therapy is dominated by a small number of invariant clones rarely found in circulating CD4+ T cells. Journal of virology. 2006;80(13):6441–57. Epub 2006/06/16. doi: 10.1128/JVI.00591-06 16775332; PubMed Central PMCID: PMC1488985.

45. Perelson AS, Essunger P, Cao Y, Vesanen M, Hurley A, Saksela K, et al. Decay characteristics of HIV-1-infected compartments during combination therapy. Nature. 1997;387(6629):188–91. Epub 1997/05/08. doi: 10.1038/387188a0 9144290.

46. Jones BR, Miller RL, Kinloch NN, Tsai O, Rigsby H, Sudderuddin H, et al. Genetic diversity, compartmentalization and age of HIV proviruses persisting in CD4+ T cell subsets during long-term combination antiretroviral therapy. Journal of virology. 2019:JVI.01786-19. Epub 2019/11/30. doi: 10.1128/JVI.01786-19 31776273.

47. Rousseau CM, Birditt BA, McKay AR, Stoddard JN, Lee TC, McLaughlin S, et al. Large-scale amplification, cloning and sequencing of near full-length HIV-1 subtype C genomes. Journal of virological methods. 2006;136(1–2):118–25. Epub 2006/05/17. doi: 10.1016/j.jviromet.2006.04.009 16701907.

48. Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol. 2010;59(3):307–21. Epub 2010/06/09. doi: 10.1093/sysbio/syq010 20525638.

49. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, et al. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012;28(12):1647–9. Epub 2012/05/01. doi: 10.1093/bioinformatics/bts199 22543367; PubMed Central PMCID: PMC3371832.

50. Rose PP, Korber BT. Detecting hypermutations in viral sequences with an emphasis on G—> A hypermutation. Bioinformatics. 2000;16(4):400–1. Epub 2000/06/27. doi: 10.1093/bioinformatics/16.4.400 10869039.

51. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33(7):1870–4. Epub 2016/03/24. doi: 10.1093/molbev/msw054 27004904.

52. Martin DP, Murrell B, Golden M, Khoosal A, Muhire B. RDP4: Detection and analysis of recombination patterns in virus genomes. Virus Evol. 2015;1(1):vev003. Epub 2015/05/26. doi: 10.1093/ve/vev003 27774277; PubMed Central PMCID: PMC5014473.

53. Suchard MA, Lemey P, Baele G, Ayres DL, Drummond AJ, Rambaut A. Bayesian phylogenetic and phylodynamic data integration using BEAST 1.10. Virus Evol. 2018;4(1):vey016. Epub 2018/06/27. doi: 10.1093/ve/vey016 29942656; PubMed Central PMCID: PMC6007674.

54. Shapiro B, Rambaut A, Drummond AJ. Choosing appropriate substitution models for the phylogenetic analysis of protein-coding sequences. Mol Biol Evol. 2006;23(1):7–9. Epub 2005/09/24. doi: 10.1093/molbev/msj021 16177232.

55. Drummond AJ, Ho SY, Phillips MJ, Rambaut A. Relaxed phylogenetics and dating with confidence. PLoS Biol. 2006;4(5):e88. Epub 2006/05/11. doi: 10.1371/journal.pbio.0040088 16683862; PubMed Central PMCID: PMC1395354.

56. Minin VN, Bloomquist EW, Suchard MA. Smooth skyride through a rough skyline: Bayesian coalescent-based inference of population dynamics. Mol Biol Evol. 2008;25(7):1459–71. Epub 2008/04/15. doi: 10.1093/molbev/msn090 18408232; PubMed Central PMCID: PMC3302198.

57. Drummond AJ, Nicholls GK, Rodrigo AG, Solomon W. Estimating mutation parameters, population history and genealogy simultaneously from temporally spaced sequence data. Genetics. 2002;161(3):1307–20. Epub 2002/07/24. 12136032; PubMed Central PMCID: PMC1462188.

58. Bouckaert R, Vaughan TG, Barido-Sottani J, Duchene S, Fourment M, Gavryushkina A, et al. BEAST 2.5: An advanced software platform for Bayesian evolutionary analysis. PLoS Comput Biol. 2019;15(4):e1006650. Epub 2019/04/09. doi: 10.1371/journal.pcbi.1006650 30958812; PubMed Central PMCID: PMC6472827.

59. Rambaut A, Drummond AJ, Xie D, Baele G, Suchard MA. Posterior Summarization in Bayesian Phylogenetics Using Tracer 1.7. Syst Biol. 2018;67(5):901–4. Epub 2018/05/03. doi: 10.1093/sysbio/syy032 29718447; PubMed Central PMCID: PMC6101584.

60. Stamatakis A. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics. 2014;30(9):1312–3. Epub 2014/01/24. doi: 10.1093/bioinformatics/btu033 24451623; PubMed Central PMCID: PMC3998144.

61. Paradis E, Schliep K. ape 5.0: an environment for modern phylogenetics and evolutionary analyses in R. Bioinformatics. 2019;35(3):526–8. Epub 2018/07/18. doi: 10.1093/bioinformatics/bty633 30016406.

62. Keele BF, Giorgi EE, Salazar-Gonzalez JF, Decker JM, Pham KT, Salazar MG, et al. Identification and characterization of transmitted and early founder virus envelopes in primary HIV-1 infection. Proceedings of the National Academy of Sciences of the United States of America. 2008;105(21):7552–7. Epub 2008/05/21. doi: 10.1073/pnas.0802203105 18490657; PubMed Central PMCID: PMC2387184.


Článek vyšel v časopise

PLOS Pathogens


2020 Číslo 6

Nejčtenější v tomto čísle

Tomuto tématu se dále věnují…


Kurzy Doporučená témata Časopisy
Přihlášení
Zapomenuté heslo

Nemáte účet?  Registrujte se

Zapomenuté heslo

Zadejte e-mailovou adresu se kterou jste vytvářel(a) účet, budou Vám na ni zaslány informace k nastavení nového hesla.

Přihlášení

Nemáte účet?  Registrujte se

VIRTUÁLNÍ ČEKÁRNA ČR Jste praktický lékař nebo pediatr? Zapojte se! Jste praktik nebo pediatr? Zapojte se!

×