Background Here, we report on the partial and full-length genomic (FLG) variability of HTLV-1 sequences from 90 well-characterized subjects, including 48 HTLV-1 asymptomatic carriers (ACs), 35 HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and 7 adult T-cell leukemia/lymphoma (ATLL) patients, using an Illumina paired-end protocol. Methods Blood samples were collected from 90 individuals, and DNA was extracted from the PBMCs to measure the proviral load and to amplify the HTLV-1 FLG from two overlapping fragments. The amplified PCR products were subjected to deep sequencing. The sequencing data were assembled, aligned, and mapped against the HTLV-1 genome with sufficient genetic resemblance and utilized for further phylogenetic analysis. Results A high-throughput sequencing-by-synthesis instrument was used to obtain an average of 3210- and 5200-fold coverage of the partial (n = 14) and FLG (n = 76) data from the HTLV-1 strains, respectively. The results based on the phylogenetic trees of consensus sequences from partial and FLGs revealed that 86 (95.5%) individuals were infected with the transcontinental sub-subtypes of the cosmopolitan subtype (aA) and that 4 individuals (4.5%) were infected with the Japanese sub-subtypes (aB). A comparison of the nucleotide and amino acids of the FLG between the three clinical settings yielded no correlation between the sequenced genotype and clinical outcomes. The evolutionary relationships among the HTLV sequences were inferred from nucleotide sequence, and the results are consistent with the hypothesis that there were multiple introductions of the transcontinental subtype in Brazil. Conclusions This study has increased the number of subtype aA full-length genomes from 8 to 81 and HTLV-1 aB from 2 to 5 sequences. The overall data confirmed that the cosmopolitan transcontinental sub-subtypes were the most prevalent in the Brazilian population. It is hoped that this valuable genomic data will add to our current understanding of the evolutionary history of this medically important virus.
References
[1]
Gessain A, Barin F, Vernant JC, Gout O, Maurs L, et al. (1985) Antibodies to human T-lymphotropic virus type-I in patients with tropical spastic paraparesis. Lancet 2: 407–410. doi: 10.1016/s0140-6736(85)92734-5
[2]
Osame M, Usuku K, Izumo S, Ijichi N, Amitani H, et al. (1986) HTLV-I associated myelopathy, a new clinical entity. Lancet 1: 1031–1032. doi: 10.1016/s0140-6736(86)91298-5
[3]
Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, et al. (1980) Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci U S A 77: 7415–7419. doi: 10.1016/b978-012448510-5/50140-0
[4]
Yoshida M, Miyoshi I, Hinuma Y (1982) Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease. Proc Natl Acad Sci U S A 79: 2031–2035. doi: 10.1073/pnas.79.6.2031
[5]
Huang YQ, Li JJ, Nicolaides A, Zhang WG, Freidman-Kien AE (1992) Fibroblast growth factor 6 gene expression in AIDS-associated Kaposi's sarcoma. Lancet 339: 1110–1111. doi: 10.1016/0140-6736(92)90702-5
[6]
Sugimoto M, Nakashima H, Watanabe S, Uyama E, Tanaka F, et al. (1987) T-lymphocyte alveolitis in HTLV-I-associated myelopathy. Lancet 2: 1220. doi: 10.1016/s0140-6736(87)91362-6
[7]
Kawai H, Saito M, Takagi M, Tsuchihashi T, Arii Y, et al. (1992) Hashimoto's thyroiditis in HTLV-I carriers. Intern Med 31: 1213–1216. doi: 10.2169/internalmedicine.31.1213
[8]
Nishioka K, Maruyama I, Sato K, Kitajima I, Nakajima Y, et al. (1989) Chronic inflammatory arthropathy associated with HTLV-I. Lancet 1: 441. doi: 10.1016/s0140-6736(89)90038-x
[9]
Edlich RF, Arnette JA, Williams FM (2000) Global epidemic of human T-cell lymphotropic virus type-I (HTLV-I). J Emerg Med 18: 109–119. doi: 10.1016/s0736-4679(99)00173-0
[10]
Matsuoka M (2003) Human T-cell leukemia virus type I and adult T-cell leukemia. Oncogene 22: 5131–5140. doi: 10.1038/sj.onc.1206551
[11]
Proietti FA, Carneiro-Proietti AB, Catalan-Soares BC, Murphy EL (2005) Global epidemiology of HTLV-I infection and associated diseases. Oncogene 24: 6058–6068. doi: 10.1038/sj.onc.1208968
[12]
Gessain A, Cassar O (2012) Epidemiological Aspects and World Distribution of HTLV-1 Infection. Front Microbiol 3: 388. doi: 10.3389/fmicb.2012.00388
[13]
Seiki M, Hattori S, Hirayama Y, Yoshida M (1983) Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci U S A 80: 3618–3622. doi: 10.1073/pnas.80.12.3618
[14]
Franchini G, Fukumoto R, Fullen JR (2003) T-cell control by human T-cell leukemia/lymphoma virus type 1. Int J Hematol 78: 280–296. doi: 10.1007/bf02983552
[15]
Verdonck K, Gonzalez E, Van Dooren S, Vandamme AM, Vanham G, et al. (2007) Human T-lymphotropic virus 1: recent knowledge about an ancient infection. Lancet Infect Dis 7: 266–281. doi: 10.1016/s1473-3099(07)70081-6
[16]
Vidal AU, Gessain A, Yoshida M, Tekaia F, Garin B, et al. (1994) Phylogenetic classification of human T cell leukaemia/lymphoma virus type I genotypes in five major molecular and geographical subtypes. J Gen Virol 75(Pt 12): 3655–3666. doi: 10.1099/0022-1317-75-12-3655
[17]
Van Dooren S, Gotuzzo E, Salemi M, Watts D, Audenaert E, et al. (1998) Evidence for a post-Columbian introduction of human T-cell lymphotropic virus [type I] [corrected] in Latin America. J Gen Virol 79(Pt 11): 2695–2708.
[18]
Galvao-Castro B, Loures L, Rodriques LG, Sereno A, Ferreira Junior OC, et al. (1997) Distribution of human T-lymphotropic virus type I among blood donors: a nationwide Brazilian study. Transfusion 37: 242–243. doi: 10.1046/j.1537-2995.1997.37297203532.x
[19]
Carneiro-Proietti AB, Sabino EC, Leao S, Salles NA, Loureiro P, et al. (2012) Human T-lymphotropic virus type 1 and type 2 seroprevalence, incidence, and residual transfusion risk among blood donors in Brazil during 2007-2009. AIDS Res Hum Retroviruses 28: 1265–1272. doi: 10.1089/aid.2011.0143
[20]
Dourado I, Alcantara LC, Barreto ML, da Gloria Teixeira M, Galvao-Castro B (2003) HTLV-I in the general population of Salvador, Brazil: a city with African ethnic and sociodemographic characteristics. J Acquir Immune Defic Syndr 34: 527–531. doi: 10.1097/00126334-200312150-00013
[21]
Catalan-Soares B, Carneiro-Proietti AB, Proietti FA (2005) Heterogeneous geographic distribution of human T-cell lymphotropic viruses I and II (HTLV-I/II): serological screening prevalence rates in blood donors from large urban areas in Brazil. Cad Saude Publica 21: 926–931. doi: 10.1590/s0102-311x2005000300027
[22]
Bimber BN, Burwitz BJ, O'Connor S, Detmer A, Gostick E, et al. (2009) Ultradeep pyrosequencing detects complex patterns of CD8+ T-lymphocyte escape in simian immunodeficiency virus-infected macaques. J Virol 83: 8247–8253. doi: 10.1128/jvi.00897-09
[23]
Bimber BN, Dudley DM, Lauck M, Becker EA, Chin EN, et al. (2010) Whole-genome characterization of human and simian immunodeficiency virus intrahost diversity by ultradeep pyrosequencing. J Virol 84: 12087–12092. doi: 10.1128/jvi.01378-10
[24]
Willerth SM, Pedro HA, Pachter L, Humeau LM, Arkin AP, et al. (2010) Development of a low bias method for characterizing viral populations using next generation sequencing technology. PLoS One 5: e13564. doi: 10.1371/journal.pone.0013564
[25]
Dudley DM, Chin EN, Bimber BN, Sanabani SS, Tarosso LF, et al. (2012) Low-cost ultra-wide genotyping using Roche/454 pyrosequencing for surveillance of HIV drug resistance. PLoS One 7: e36494. doi: 10.1371/journal.pone.0036494
[26]
Wang C, Mitsuya Y, Gharizadeh B, Ronaghi M, Shafer RW (2007) Characterization of mutation spectra with ultra-deep pyrosequencing: application to HIV-1 drug resistance. Genome Res 17: 1195–1201. doi: 10.1101/gr.6468307
[27]
Osame M (1990) Review of WHO Kagoshima Meeting and diagnostic guidelines for HAM/TSP.In Human Retrovirology: HTLV. Edited by Blattner W. New York: Raven. 191–197.
[28]
Heneine W, Khabbaz RF, Lal RB, Kaplan JE (1992) Sensitive and specific polymerase chain reaction assays for diagnosis of human T-cell lymphotropic virus type I (HTLV-I) and HTLV-II infections in HTLV-I/II-seropositive individuals. J Clin Microbiol 30: 1605–1607.
[29]
Naderi M, Paryan M, Azadmanesh K, Rafatpanah H, Rezvan H, et al. (2012) Design and development of a quantitative real time PCR assay for monitoring of HTLV-1 provirus in whole blood. J Clin Virol 53: 302–307. doi: 10.1016/j.jcv.2011.12.033
[30]
Katoh K, Misawa K, Kuma K, Miyata T (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 30: 3059–3066. doi: 10.1093/nar/gkf436
[31]
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, et al. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731–2739. doi: 10.1093/molbev/msr121
[32]
Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, et al. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 61: 539–542.
[33]
Han MV, Zmasek CM (2009) phyloXML: XML for evolutionary biology and comparative genomics. BMC Bioinformatics 10: 356. doi: 10.1186/1471-2105-10-356
[34]
Furukawa Y, Yamashita M, Usuku K, Izumo S, Nakagawa M, et al. (2000) Phylogenetic subgroups of human T cell lymphotropic virus (HTLV) type I in the tax gene and their association with different risks for HTLV-I-associated myelopathy/tropical spastic paraparesis. J Infect Dis 182: 1343–1349. doi: 10.1086/315897
[35]
Kashima S, Alcantara LC, Takayanagui OM, Cunha MA, Castro BG, et al. (2006) Distribution of human T cell lymphotropic virus type 1 (HTLV-1) subtypes in Brazil: genetic characterization of LTR and tax region. AIDS Res Hum Retroviruses 22: 953–959. doi: 10.1089/aid.2006.22.953
[36]
Segurado AA, Biasutti C, Zeigler R, Rodrigues C, Damas CD, et al. (2002) Identification of human T-lymphotropic virus type I (HTLV-I) subtypes using restricted fragment length polymorphism in a cohort of asymptomatic carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis from Sao Paulo, Brazil. Mem Inst Oswaldo Cruz 97: 329–333. doi: 10.1590/s0074-02762002000300009
[37]
Magri MC, Brigido LF, Rodrigues R, Morimoto HK, Ferreira JL, et al. (2012) Phylogenetic and similarity analysis of HTLV-1 isolates from HIV-coinfected patients from the south and southeast regions of Brazil. AIDS Res Hum Retroviruses 28: 110–114. doi: 10.1089/aid.2011.0117
[38]
Neto WK, Da-Costa AC, de Oliveira AC, Martinez VP, Nukui Y, et al. (2011) Correlation between LTR point mutations and proviral load levels among human T cell lymphotropic virus type 1 (HTLV-1) asymptomatic carriers. Virol J 8: 535. doi: 10.1186/1743-422x-8-535
[39]
Leitner T (2002) The molecular epidemiology of human viruses. Berlin, Germany: Springer Science and Business Media.
[40]
Komurian F, Pelloquin F, de The G (1991) In vivo genomic variability of human T-cell leukemia virus type I depends more upon geography than upon pathologies. J Virol 65: 3770–3778.
[41]
Longo DL, Gelmann EP, Cossman J, Young RA, Gallo RC, et al. (1984) Isolation of HTLV-transformed B-lymphocyte clone from a patient with HTLV-associated adult T-cell leukaemia. Nature 310: 505–506. doi: 10.1038/310505a0
[42]
Mahieux R, Ibrahim F, Mauclere P, Herve V, Michel P, et al. (1997) Molecular epidemiology of 58 new African human T-cell leukemia virus type 1 (HTLV-1) strains: identification of a new and distinct HTLV-1 molecular subtype in Central Africa and in Pygmies. J Virol 71: 1317–1333.
[43]
Alcantara LC, de Oliveira T, Gordon M, Pybus O, Mascarenhas RE, et al. (2006) Tracing the origin of Brazilian HTLV-1 as determined by analysis of host and viral genes. AIDS 20: 780–782. doi: 10.1097/01.aids.0000216383.14808.13
[44]
Mota AC, Van Dooren S, Fernandes FM, Pereira SA, Queiroz AT, et al. (2007) The close relationship between South African and Latin American HTLV type 1 strains corroborated in a molecular epidemiological study of the HTLV type 1 isolates from a blood donor cohort. AIDS Res Hum Retroviruses 23: 503–507. doi: 10.1089/aid.2006.0203
[45]
Eirin ME, Dilernia DA, Berini CA, Jones LR, Pando MA, et al. (2008) Divergent strains of human T-lymphotropic virus type 1 (HTLV-1) within the Cosmopolitan subtype in Argentina. AIDS Res Hum Retroviruses 24: 1237–1244. doi: 10.1089/aid.2008.0024
[46]
Balcazar N, Sanchez GI, Garcia-Vallejo F (2003) Sequence and phylogenetic analysis of human T cell lymphotropic virus type 1 from Tumaco, Colombia. Mem Inst Oswaldo Cruz 98: 641–648. doi: 10.1590/s0074-02762003000500010
[47]
Song KJ, Nerurkar VR, Pereira-Cortez AJ, Yamamoto M, Taguchi H, et al. (1995) Sequence and phylogenetic analyses of human T cell lymphotropic virus type 1 from a Brazilian woman with adult T cell leukemia: comparison with virus strains from South America and the Caribbean basin. Am J Trop Med Hyg 52: 101–108.
[48]
Yamashita M, Ishida T, Ohkura S, Miura T, Hayami M (2001) Phylogenetic characterization of a new HTLV type 1 from the Ainu in Japan. AIDS Res Hum Retroviruses 17: 783–787. doi: 10.1089/088922201750237068