In vivo expression of the HBZ gene of HTLV-1 correlates with proviral load, inflammatory markers and disease severity in HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP)
© Saito et al; licensee BioMed Central Ltd. 2009
Received: 27 November 2008
Accepted: 19 February 2009
Published: 19 February 2009
Recently, human T-cell leukemia virus type 1 (HTLV-1) basic leucine zipper factor (HBZ), encoded from a minus strand mRNA was discovered and was suggested to play an important role in adult T cell leukemia (ATL) development. However, there have been no reports on the role of HBZ in patients with HTLV-1 associated inflammatory diseases.
We quantified the HBZ and tax mRNA expression levels in peripheral blood from 56 HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients, 10 ATL patients, 38 healthy asymptomatic carriers (HCs) and 20 normal uninfected controls, as well as human leukemic T-cell lines and HTLV-1-infected T-cell lines, and the data were correlated with clinical parameters. The spliced HBZ gene was transcribed in all HTLV-1-infected individuals examined, whereas tax mRNA was not transcribed in significant numbers of subjects in the same groups. Although the amount of HBZ mRNA expression was highest in ATL, medium in HAM/TSP, and lowest in HCs, with statistical significance, neither tax nor the HBZ mRNA expression per HTLV-1-infected cell differed significantly between each clinical group. The HTLV-1 HBZ, but not tax mRNA load, positively correlated with disease severity and with neopterin concentration in the cerebrospinal fluid of HAM/TSP patients. Furthermore, HBZ mRNA expression per HTLV-1-infected cell was decreased after successful immunomodulatory treatment for HAM/TSP.
These findings suggest that in vivo expression of HBZ plays a role in HAM/TSP pathogenesis.
Human T-cell lymphotropic virus type 1 (HTLV-1) is a replication-competent human retrovirus [1, 2] which is associated with adult T-cell leukemia (ATL) [3, 4] and with a slowly progressive neurological disorder HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) [5, 6]. In HTLV-1 infection, approximately 5% develop ATL  and another 2%-3% develop chronic inflammatory diseases involving the central nervous system (HAM/TSP), the eyes , the lungs , the joints , or the skeletal muscles ; most infected individuals, however, remain healthy in their lifetime (healthy asymptomatic carriers: HCs). Although the factors that cause these different manifestations of HTLV-1 infection are not fully understood, previous population association studies suggested that both viral and host genetic factors influence the outcome of infection .
Among several HTLV-1 genes, a transcriptional activator Tax encoded in the pX region is thought to play a central role in immortalization, oncogenesis and inflammation through its pleiotropic activity . In HAM/TSP patients, it has been reported that several cytokines, chemokines and matrix metalloproteinases transactivated by Tax protein such as tumor necrosis factor-α (TNF-α) , monocyte chemoattractant protein-1 (MCP-1)  and matrix metalloproteinase (MMP)-9  are overexpressed in the infiltrating mononuclear cells in the patients' spinal cords. In addition, a previous report from the United States suggested that the level of HTLV-1 tax mRNA expression in HTLV-1-infected cells (mRNA/DNA ratio) was significantly higher in HAM/TSP patients than HCs, and this finding correlated with the HTLV-1 proviral load, Tax-specific CD8+ T cell frequency and disease severity of the patients . A report from Japan also indicated that HTLV-1 tax mRNA expression was higher in HAM/TSP than HCs, although the mRNA/DNA ratio was similar between both groups . These results suggest an important role of Tax in the induction of HAM/TSP.
It has been reported that among fresh leukemic cells isolated from ATL patients, about 60% of cases do not express the tax transcript . In tax transgenic mouse models, the mice develop a wide range of tumors such as neurofibrosarcomas, mesenchymal tumors, and mammary adenomas, or even skeletal abnormalities including osteolytic bone metastases [20–27]; however, no leukemias or lymphomas were identified except in three models, which used respectively the granzyme B promoter , Lck proximal promoter  and Lck distal promoter . These findings suggest that Tax is required for malignant transformation but not essential for the maintenance of leukemic cells in vivo. Recently, a novel basic leucine zipper protein encoded by the complementary strand of the HTLV-1 genome, named HTLV-1 basic leucine zipper factor (HBZ), was characterized . HBZ is expressed in all ATL cells , promotes proliferation of T-lymphocytes in its RNA form , suppresses Tax-mediated transactivation through the 5' LTR [31, 33], promotes CD4+ T-lymphocyte proliferation in transgenic mice , and enhances infectivity and persistence in HTLV-1-inoculated rabbits .
In this study, we investigated whether HTLV-1 HBZ mRNA expression is associated with clinical and laboratory markers reported in HAM/TSP patients, including HTLV-1 proviral load, neopterin concentration in cerebrospinal fluid (CSF), and motor disability score. In addition, to confirm the previous observations [17, 18], we have also investigated the tax mRNA expression in ATL patients, HAM/TSP patients, and HCs by using the same technology but in a larger number of subjects.
Patients and cells
Human leukemic T-cell lines (Jurkat, MOLT-4, and CEM) and HTLV-1-infected T-cell lines (C5/MJ, SLB1, HUT102, MT-1, MT-2, and MT-4) were cultured in RPMI 1640 medium supplemented with 10% FCS. The diagnosis of HAM/TSP was done in accordance with World Health Organization criteria . The diagnosis of ATL was made on the basis of clinical features, hematological characteristics, serum antibodies against HTLV-1 antigens, and detection of the HTLV-1 viral genome inserted into leukemia cells by Southern blot hybridization. All the PBMC samples used in this study were collected prior to treatment by a Histopaque-1077 (Sigma) density gradient centrifugation, washed and stored in liquid nitrogen until use. This research was approved by the institutional review boards of the authors' institutions, and informed consent was obtained from all individuals.
Quantification of HTLV-1 proviral load, tax and HBZ mRNA expression, anti-HTLV-1 antibody titers and neopterin concentration in cerebrospinal fluid
RNA was extracted from PBMCs using RNeasy Mini Kit with on-column DNase digestion (QIAGEN, Tokyo, Japan) according to the manufacturer's instructions. Complementary DNA (cDNA) was synthesized using TaqMan Gold RT-PCR Kit (Applied Biosystems, Tokyo, Japan). For cDNA synthesis from extracted mRNA, 2 μg total RNA, 10 μl 10×TaqMan RT buffer, 22 μl MgCl2 (25 mM), 20 μl dNTPs mixture (at a final concentration of 500 μM each), 5 μl random hexamers (50 μM), 2 μl RNase inhibitor (20 U/μl), and 2.5 μl (50 U/μl) Moloney murine leukemia virus reverse transcriptase were added to a total volume of 100 μl. Samples were incubated at 25°C for 10 minutes and 48°C for 30 minutes, and reactions were stopped by heating to 95°C for 5 minutes. Genomic DNA was extracted from the frozen PBMCs by QIAamp blood kit (QIAGEN, Tokyo, Japan). We, then, carried out a real time quantitative PCR using ABI Prism 7900 HT Fast Real-Time PCR System (Applied Biosystems) to examine the HTLV-1 proviral load  and tax mRNA expression  in PBMCs or HTLV-1 infected cell lines as reported previously. The amount of the HTLV-1 proviral load was calculated using β-actin as an internal control through the following formula: copy number of HTLV-1 tax per cell = [(copy number of tax)/(copy number of β-actin/2)]. The sequences of primers for HTLV-1 provirus were as follows: 5'-CAA ACC GTC AAG CAC AGC TT-3' and 5'-TCT CCA AAC ACG TAG ACT GGG T-3', and the probe was 5'-TTC CCA GGG TTT GGA CAG AGT CTT CT-3'. HBZ mRNA expression levels were also quantified by real time quantitative PCR using the same method for tax mRNA . Namely, serially diluted cDNA from HTLV-1 infected MT-2 cells was used for generating standard curves for the value of HTLV-1 tax or HBZ mRNA and hypoxanthine ribosyl transferase (HPRT) mRNA, and the relative HTLV-1 tax or HBZ mRNA load was calculated by the following formula: HTLV-1 tax mRNA load = value of tax/value of HPRT. HTLV-1 HBZ mRNA load = value of HBZ/value of HPRT. We used aliquots of the same standard MT-2 cDNA preparation for all assays and the correlation values of standard curves were always more than 99%. The sequences of primers for tax mRNA detection were as follows: 5'-ATC CCG TGG AGA CTC CTC AA-3' and 5'-ATC CCG TGG AGA CTC CTC AA-3', and the probe was 5'-TCC AAC ACC ATG GCC CAC TTC CC-3'. The sequences of primers for HBZ mRNA detection were as follows: 5'-AGA ACG CGA CTC AAC CGG-3' and 5'-TGA CAC AGG CAA GCA TCG A-3', and the probe was 5'-TGG ATG GCG GCC TCA GGG CT-3'. As the probes for tax and HBZ mRNA surrounded the splice junction site of each mRNA, we detected HBZ splicing isoform, which is the most abundant HBZ transcript and contributed significantly to HBZ protein synthesis [37–39], but not unspliced form in this study. We used the HPRT primers and probe set (Applied Biosystems) for internal calibration. The tax and HBZ probes were labeled with fluorescent 6-carboxyfluorescein (FAM) (reporter) at the 5' end and fluorescent 6-carboxy tetramethyl rhodamine (TAMRA) (quencher) at the 3' end. All assays were performed in triplicate. The sensitivity of our real-time RT-PCR assay was determined using MT-2 cells diluted serially with PBMCs from a healthy uninfected donor. The HTLV-1 mRNA signal (both tax and HBZ) could be detected in a dose-dependent manner with a sensitivity limit as low as one MT-2 cell in 106 PBMCs. Neopterin levels were evaluated by HPLC with fluorometric detection methods as described previously . Serum HTLV-1 antibody titers were determined by a particle agglutination method (Serodia-HTLV-1®, Fujirebio, Japan).
Motor dysfunction seen in HAM/TSP patients was evaluated by clinical neurologists according to the Osame Motor Disability Score (OMDS) , which grades motor dysfunction from zero (normal walking and running) to 13 (complete bedridden) as follows: 1 = normal gait but runs slow; 2 = abnormal gait; 3 = abnormal gait and unable to run; 4 = need support while using stairs; 5 = need one hand support in walking; 6 = need two hands support in walking; 7 = need two hands support in walking but is limited to 10 m; 8 = need two hands support in walking but is limited to 5 m; 9 = unable to walk but able to crawl on hands and knees; 10 = crawls with hands; 11 = unable to crawl but can turn sideways in bed; 12 = unable to turn sideways but can move the toes. We have used OMDS throughout our previous studies [41–43] because this is a neurological measure of disability weighted toward ambulation and was specifically developed to evaluate motor dysfunction seen in HAM/TSP patients. It is therefore more suitable for evaluating HAM/TSP motor symptoms than the widely used EDSS . The laboratory data were examined by an investigator who was not involved in the patients' clinical care, and the neurologists who made the clinical evaluation did not have access to the laboratory data.
The Mann-Whitney U test was used to compare data between two groups. Correlations between variables were examined by Spearman rank correlation analysis. Values of p < 0.05 were considered statistically significant.
HTLV-1 tax and HBZ mRNA load in HAM/TSP, ATL and HCs
HTLV-1 mRNA load, proviral load and mRNA/DNA ratio in HTLV-1 – infected individuals and T-cell lines.
Comparison of HTLV-1 tax and HBZ mRNA load with HTLV-1 proviral load
Comparison of HBZ mRNA load with tax mRNA load among HTLV-1 infected individuals in different clinical status
Correlation of HTLV-1 HBZ mRNA load with CSF neopterin concentration and disease severity in HAM/TSP patients
Results of rank correlation test between clinical and virological parameters.
Neopterin in CSF
HBZ mRNA load and HBZ mRNA/DNA ratio in PBMCs was decreased in HAM/TSP patients after effective IFN-α treatment
In this study, we demonstrated that there was a statistically significant difference in the HTLV-1 HBZ mRNA load, but not tax mRNA load, in PBMCs between HAM/TSP patients and HCs. This is probably because tax mRNA was not expressed in significant numbers of individuals tested (60.7% of HAM/TSP patients, 34 out of 56; 71.1% of HCs, 27 out of 38; 30.0% of ATL patients, 3 out of 10), whereas HTLV-1 HBZ mRNA was expressed in all the infected individuals tested. There was also a statistically significant correlation between HTLV-1 HBZ mRNA load and HTLV-1 proviral load both in HAM/TSP patients and HCs, whereas tax mRNA load correlated with the HTLV-1 proviral load only in HCs but not in HAM/TSP patients. Recently, Usui et al. reported a similar observation . Namely, HBZ spliced isoform mRNA was detectable in samples from most HCs and ATL patients, and was significantly correlated with the HTLV-1 proviral load. These results indicate that the regulation of HBZ mRNA expression is different from that of tax mRNA. It seems likely that HBZ mRNA is near-equally expressed by all provirus-positive cells despite different clinical status, while tax mRNA expression levels are variable in different clinical status.
When HTLV-1 tax or HBZ mRNA load was adjusted with HTLV-1 proviral DNA load (i.e. calculate mRNA/DNA ratio), the amount of tax and HBZ mRNA expressed per provirus was not significantly different between HAM/TSP patients and HCs, suggesting that the higher HTLV-1 proviral load seen in HAM/TSP patients caused higher HTLV-1 HBZ mRNA expression. This is consistent with our previous study using different methods for mRNA and DNA quantification , but differed from a previous American study using exactly the same methods, which showed significantly higher mRNA/DNA ratio in HAM/TSP patients than HCs . In contrast to the previous study, which showed significant correlation between disease severity in HAM/TSP patients and both HTLV-1 tax mRNA load and mRNA/DNA ratio , we could not find such a correlation between clinical parameters of HAM/TSP patients including disease severity and both HTLV-1 tax mRNA load and mRNA/DNA ratio (Table 2). As we have already confirmed and reported the same levels of Tax protein expression in HTLV-1-infected PBMCs between HAM/TSP patients and HCs in the same cohort , the observed discrepancy may be due to the differences of a number of host genetic and virologic factors in HTLV-1 infected individuals, including differences in HLA haplotypes [51–53], differences in the amount of soluble suppressive factors and CD8+ T-cell responses, and differences in HTLV-1 tax genomic sequences . As a recent report indicated that HTLV-I infection was associated with activated T-cell immunity in Jamaicans but with diminished T-cell immunity in Japanese persons , the interaction between different genes and/or environmental factors is also likely to contribute to the observed differences between the two populations. Namely, genetic resistance to infectious diseases that is formed by complex host genetic effects might be complicated further by pathogen diversity and environmental factors.
Another important observation is that the amount of HTLV-1 HBZ mRNA expression per provirus was more than a thousand times higher than tax mRNA expression both in HAM/TSP patients and HCs. Surprisingly, the amount of HTLV-1 HBZ mRNA expression per provirus was even higher in HTLV-1-infected PBMCs than in infected cell lines, whereas tax mRNA expression was significantly higher in cell lines than infected PBMCs. Since HBZ suppresses Tax-mediated viral transcription , the abundant expression of HBZ mRNA in HTLV-1-infected PBMCs will be one of the molecular mechanisms involved in viral latency by suppressing HTLV-1 transcription and Tax expression, which may be a significant advantage to the virus in the infected cell by preventing its detection through a CTL response. Since we and others  found that down-regulation of tax mRNA (higher HBZ mRNA/tax mRNA ratio) was characteristic of primary ATL cells, imbalanced expression between HBZ and tax may induce the outgrowth of HTLV-1-transformed T cell and increase the risk of ATL, which is associated with a Tax-low or -negative phenotype.
We also found that the HTLV-1 HBZ mRNA load significantly correlated with the neopterin concentrations in CSF of HAM/TSP patients. Since neopterin levels in CSF have been used as an immunologic marker for monitoring disease activity and treatment efficacy of HAM/TSP [40, 42, 56], the quantitative analysis of HTLV-1 HBZ mRNA might also be used to monitor HAM/TSP disease activity. As expected, motor dysfunction of HAM/TSP patients evaluated by the OMDS score significantly correlated with HTLV-1 HBZ mRNA load (P = 0.023) but not with HTLV-1 tax mRNA load (P = 0.401). The correlation between HBZ mRNA load and two independent clinical parameters reflecting disease activities strongly suggest its stronger relevance than both tax mRNA and proviral load for HAM/TSP pathogenesis. This is further supported by the data that both HBZ mRNA load and HBZ mRNA/DNA ratio were decreased in HAM/TSP patients after effective IFN-α treatment. Collectively, our results suggest that higher HTLV-1 HBZ mRNA load may have relative prognostic value for the assessment of disease progression and could also be used as a surrogate marker to predict long-term outcome in HAM/TSP patients.
In summary, we showed that spliced HBZ gene was transcribed in all the HTLV-1 infected individuals examined, whereas tax mRNA was not transcribed in more than half in the same groups. Moreover, our data demonstrated a significant correlation between HTLV-1 HBZ mRNA load and HTLV-1 proviral load, neopterin concentrations in CSF and motor disability seen in HAM/TSP patients, indicating that HTLV-1 HBZ mRNA load may be a valid predictor of disease progression. Our present findings suggest that HTLV-1 HBZ mRNA expression plays a role not only in ATL, but also in the pathogenesis of the HTLV-1-associated inflammatory disease HAM/TSP.
We are grateful to the staff and blood donors of Kagoshima University Hospital. We also thank Dr. Ryuji Kubota for providing the clinical samples, Prof. Masahiro Fujii of Niigata University for the gift of HTLV-1-infected T-cell lines (C5/MJ, SLB1, and MT-4), and Ms. Sumie Saito of Kanazawa Medical University for technical assistance. This work was supported by the Ministry of Health, Labor and Welfare, Japan (Neuroimmunological Disease Research Committee Grant to Y.O.); Takeda Science Foundation (to M.S.); Kanazawa Medical University (Grants H2007-11, H2008-11, C2008-2, and S2008-8 to M.S.).
- Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC: 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 USA. 1980, 77: 7415-7419. 10.1073/pnas.77.12.7415.PubMed CentralView ArticlePubMedGoogle Scholar
- Yoshida M, Miyoshi I, Hinuma Y: Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease. Proc Natl Acad Sci USA. 1982, 79: 2031-2035. 10.1073/pnas.79.6.2031.PubMed CentralView ArticlePubMedGoogle Scholar
- Hinuma Y, Nagata K, Hanaoka M, Nakai M, Matsumoto T, Kinoshita KI, Shirakawa S, Miyoshi I: Adult T-cell leukemia: antigen in an ATL cell line and detection of antibodies to the antigen in human sera. Proc Natl Acad Sci USA. 1981, 78: 6476-6480. 10.1073/pnas.78.10.6476.PubMed CentralView ArticlePubMedGoogle Scholar
- Yoshida M, Seiki M, Yamaguchi K, Takatsuki K: Monoclonal integration of human T-cell leukemia provirus in all primary tumors of adult T-cell leukemia suggests causative role of human T-cell leukemia virus in the disease. Proc Natl Acad Sci USA. 1984, 81: 2534-2537. 10.1073/pnas.81.8.2534.PubMed CentralView ArticlePubMedGoogle Scholar
- Gessain A, Barin F, Vernant JC, Gout O, Maurs L, Calender A, de The G: Antibodies to human T-lymphotropic virus type-I in patients with tropical spastic paraparesis. Lancet. 1985, 2: 407-410. 10.1016/S0140-6736(85)92734-5.View ArticlePubMedGoogle Scholar
- Osame M, Usuku K, Izumo S, Ijichi N, Amitani H, Igata A, Matsumoto M, Tara M: HTLV-I associated myelopathy, a new clinical entity. Lancet. 1986, 1: 1031-1032. 10.1016/S0140-6736(86)91298-5.View ArticlePubMedGoogle Scholar
- Arisawa K, Soda M, Endo S, Kurokawa K, Katamine S, Shimokawa I, Koba T, Takahashi T, Saito H, Doi H, Shirahama S: Evaluation of adult T-cell leukemia/lymphoma incidence and its impact on non-Hodgkin lymphoma incidence in southwestern Japan. Int J Cancer. 2000, 85: 319-324. 10.1002/(SICI)1097-0215(20000201)85:3<319::AID-IJC4>3.0.CO;2-B.View ArticlePubMedGoogle Scholar
- Mochizuki M, Watanabe T, Yamaguchi K, Takatsuki K, Yoshimura K, Shirao M, Nakashima S, Mori S, Araki S, Miyata N: HTLV-I uveitis: a distinct clinical entity caused by HTLV-I. Jpn J Cancer Res. 1992, 83: 236-239.View ArticlePubMedGoogle Scholar
- Sugimoto M, Nakashima H, Watanabe S, Uyama E, Tanaka F, Ando M, Araki S, Kawasaki S: T-lymphocyte alveolitis in HTLV-I-associated myelopathy. Lancet. 1987, 2: 1220-10.1016/S0140-6736(87)91362-6.View ArticlePubMedGoogle Scholar
- Nishioka K, Maruyama I, Sato K, Kitajima I, Nakajima Y, Osame M: Chronic inflammatory arthropathy associated with HTLV-I. Lancet. 1989, 1: 441-10.1016/S0140-6736(89)90038-X.View ArticlePubMedGoogle Scholar
- Higuchi I, Montemayor ES, Izumo S, Inose M, Osame M: Immunohistochemical characteristics of polymyositis in patients with HTLV-I-associated myelopathy and HTLV-I carriers. Muscle Nerve. 1993, 16: 472-476. 10.1002/mus.880160507.View ArticlePubMedGoogle Scholar
- Bangham CR, Osame M: Cellular immune response to HTLV-1. Oncogene. 2005, 24: 6035-6046. 10.1038/sj.onc.1208970.View ArticlePubMedGoogle Scholar
- Yoshida M: Multiple viral strategies of HTLV-1 for dysregulation of cell growth control. Annu Rev Immunol. 2001, 19: 475-496. 10.1146/annurev.immunol.19.1.475.View ArticlePubMedGoogle Scholar
- Umehara F, Izumo S, Ronquillo AT, Matsumuro K, Sato E, Osame M: Cytokine expression in the spinal cord lesions in HTLV-I-associated myelopathy. J Neuropathol Exp Neurol. 1994, 53: 72-77. 10.1097/00005072-199401000-00009.View ArticlePubMedGoogle Scholar
- Umehara F, Izumo S, Takeya M, Takahashi K, Sato E, Osame M: Expression of adhesion molecules and monocyte chemoattractant protein -1 (MCP-1) in the spinal cord lesions in HTLV-I-associated myelopathy. Acta Neuropathol (Berl). 1996, 91: 343-350. 10.1007/s004010050435.View ArticleGoogle Scholar
- Umehara F, Okada Y, Fujimoto N, Abe M, Izumo S, Osame M: Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in HTLV-I-associated myelopathy. J Neuropathol Exp Neurol. 1998, 57: 839-849. 10.1097/00005072-199809000-00005.View ArticlePubMedGoogle Scholar
- Yamano Y, Nagai M, Brennan M, Mora CA, Soldan SS, Tomaru U, Takenouchi N, Izumo S, Osame M, Jacobson S: Correlation of human T-cell lymphotropic virus type 1 (HTLV-1) mRNA with proviral DNA load, virus-specific CD8(+) T cells, and disease severity in HTLV-1-associated myelopathy (HAM/TSP). Blood. 2002, 99: 88-94. 10.1182/blood.V99.1.88.View ArticlePubMedGoogle Scholar
- Furukawa Y, Osame M, Kubota R, Tara M, Yoshida M: Human T-cell leukemia virus type-1 (HTLV-1) Tax is expressed at the same level in infected cells of HTLV-1-associated myelopathy or tropical spastic paraparesis patients as in asymptomatic carriers but at a lower level in adult T-cell leukemia cells. Blood. 1995, 85: 1865-1870.PubMedGoogle Scholar
- Matsuoka M: Human T-cell leukemia virus type I (HTLV-I) infection and the onset of adult T-cell leukemia (ATL). Retrovirology. 2005, 2: 27-10.1186/1742-4690-2-27.PubMed CentralView ArticlePubMedGoogle Scholar
- Hinrichs SH, Nerenberg M, Reynolds RK, Khoury G, Jay G: A transgenic mouse model for human neurofibromatosis. Science. 1987, 237: 1340-1343. 10.1126/science.2888191.View ArticlePubMedGoogle Scholar
- Nerenberg M, Hinrichs SH, Reynolds RK, Khoury G, Jay G: The tat gene of human T-lymphotropic virus type 1 induces mesenchymal tumors in transgenic mice. Science. 1987, 237: 1324-1329. 10.1126/science.2888190.View ArticlePubMedGoogle Scholar
- Green JE, Hinrichs SH, Vogel J, Jay G: Exocrinopathy resembling Sjogren's syndrome in HTLV-1 tax transgenic mice. Nature. 1989, 341: 72-74. 10.1038/341072a0.View ArticlePubMedGoogle Scholar
- Iwakura Y, Tosu M, Yoshida E, Takiguchi M, Sato K, Kitajima I, Nishioka K, Yamamoto K, Takeda T, Hatanaka M, et al: Induction of inflammatory arthropathy resembling rheumatoid arthritis in mice transgenic for HTLV-I. Science. 1991, 253: 1026-1028. 10.1126/science.1887217.View ArticlePubMedGoogle Scholar
- Ruddle NH, Li CB, Horne WC, Santiago P, Troiano N, Jay G, Horowitz M, Baron R: Mice transgenic for HTLV-I LTR-tax exhibit tax expression in bone, skeletal alterations, and high bone turnover. Virology. 1993, 197: 196-204. 10.1006/viro.1993.1580.View ArticlePubMedGoogle Scholar
- Hall AP, Irvine J, Blyth K, Cameron ER, Onions DE, Campbell ME: Tumours derived from HTLV-I tax transgenic mice are characterized by enhanced levels of apoptosis and oncogene expression. J Pathol. 1998, 186: 209-214. 10.1002/(SICI)1096-9896(1998100)186:2<209::AID-PATH162>3.0.CO;2-I.View ArticlePubMedGoogle Scholar
- Gao L, Deng H, Zhao H, Hirbe A, Harding J, Ratner L, Weilbaecher K: HTLV-1 Tax transgenic mice develop spontaneous osteolytic bone metastases prevented by osteoclast inhibition. Blood. 2005, 106: 4294-4302. 10.1182/blood-2005-04-1730.PubMed CentralView ArticlePubMedGoogle Scholar
- Furuta Y, Aizawa S, Suda Y, Ikawa Y, Kishimoto H, Asano Y, Tada T, Hikikoshi A, Yoshida M, Seiki M: Thymic atrophy characteristic in transgenic mice that harbor pX genes of human T-cell leukemia virus type I. J Virol. 1989, 63: 3185-3189.PubMed CentralPubMedGoogle Scholar
- Grossman WJ, Kimata JT, Wong FH, Zutter M, Ley TJ, Ratner L: Development of leukemia in mice transgenic for the tax gene of human T-cell leukemia virus type I. Proc Natl Acad Sci USA. 1995, 92: 1057-1061. 10.1073/pnas.92.4.1057.PubMed CentralView ArticlePubMedGoogle Scholar
- Hasegawa H, Sawa H, Lewis MJ, Orba Y, Sheehy N, Yamamoto Y, Ichinohe T, Tsunetsugu-Yokota Y, Katano H, Takahashi H, Matsuda J, Sata T, Kurata T, Nagashima K, Hall WW: Thymus-derived leukemia-lymphoma in mice transgenic for the Tax gene of human T-lymphotropic virus type I. Nat Med. 2006, 12: 466-472. 10.1038/nm1389.View ArticlePubMedGoogle Scholar
- Ohsugi T, Kumasaka T, Okada S, Urano T: The Tax protein of HTLV-1 promotes oncogenesis in not only immature T cells but also mature T cells. Nat Med. 2007, 13: 527-528. 10.1038/nm0507-527.View ArticlePubMedGoogle Scholar
- Gaudray G, Gachon F, Basbous J, Biard-Piechaczyk M, Devaux C, Mesnard JM: The complementary strand of the human T-cell leukemia virus type 1 RNA genome encodes a bZIP transcription factor that down-regulates viral transcription. J Virol. 2002, 76: 12813-12822. 10.1128/JVI.76.24.12813-12822.2002.PubMed CentralView ArticlePubMedGoogle Scholar
- Satou Y, Yasunaga J, Yoshida M, Matsuoka M: HTLV-I basic leucine zipper factor gene mRNA supports proliferation of adult T cell leukemia cells. Proc Natl Acad Sci USA. 2006, 103: 720-725. 10.1073/pnas.0507631103.PubMed CentralView ArticlePubMedGoogle Scholar
- Basbous J, Arpin C, Gaudray G, Piechaczyk M, Devaux C, Mesnard JM: The HBZ factor of human T-cell leukemia virus type I dimerizes with transcription factors JunB and c-Jun and modulates their transcriptional activity. J Biol Chem. 2003, 278: 43620-43627. 10.1074/jbc.M307275200.View ArticlePubMedGoogle Scholar
- Arnold J, Yamamoto B, Li M, Phipps AJ, Younis I, Lairmore MD, Green PL: Enhancement of infectivity and persistence in vivo by HBZ, a natural antisense coded protein of HTLV-1. Blood. 2006, 107: 3976-3982. 10.1182/blood-2005-11-4551.PubMed CentralView ArticlePubMedGoogle Scholar
- Osame M: Review of WHO Kagoshima meeting and diagnostic guidelines for HAM/TSP. 1990, New York: Raven PressGoogle Scholar
- Nagai M, Usuku K, Matsumoto W, Kodama D, Takenouchi N, Moritoyo T, Hashiguchi S, Ichinose M, Bangham CR, Izumo S, Osame M: Analysis of HTLV-I proviral load in 202 HAM/TSP patients and 243 asymptomatic HTLV-I carriers: high proviral load strongly predisposes to HAM/TSP. J Neurovirol. 1998, 4: 586-593. 10.3109/13550289809114225.View ArticlePubMedGoogle Scholar
- Usui T, Yanagihara K, Tsukasaki K, Murata K, Hasegawa H, Yamada Y, Kamihira S: Characteristic expression of HTLV-1 basic zipper factor (HBZ) transcripts in HTLV-1 provirus-positive cells. Retrovirology. 2008, 5: 34-10.1186/1742-4690-5-34.PubMed CentralView ArticlePubMedGoogle Scholar
- Murata K, Hayashibara T, Sugahara K, Uemura A, Yamaguchi T, Harasawa H, Hasegawa H, Tsuruda K, Okazaki T, Koji T, Miyanishi T, Yamada Y, Kamihira S: A novel alternative splicing isoform of human T-cell leukemia virus type 1 bZIP factor (HBZ-SI) targets distinct subnuclear localization. J Virol. 2006, 80: 2495-2505. 10.1128/JVI.80.5.2495-2505.2006.PubMed CentralView ArticlePubMedGoogle Scholar
- Cavanagh MH, Landry S, Audet B, Arpin-Andre C, Hivin P, Pare ME, Thete J, Wattel E, Marriott SJ, Mesnard JM, Barbeau B: HTLV-I antisense transcripts initiating in the 3'LTR are alternatively spliced and polyadenylated. Retrovirology. 2006, 3: 15-10.1186/1742-4690-3-15.PubMed CentralView ArticlePubMedGoogle Scholar
- Nomoto M, Utatsu Y, Soejima Y, Osame M: Neopterin in cerebrospinal fluid: a useful marker for diagnosis of HTLV-I-associated myelopathy/tropical spastic paraparesis. Neurology. 1991, 41: 457-View ArticlePubMedGoogle Scholar
- Izumo S, Goto I, Itoyama Y, Okajima T, Watanabe S, Kuroda Y, Araki S, Mori M, Nagataki S, Matsukura S, Akamine T, Nakagawa M, Yamamoto I, Osame M: Interferon-alpha is effective in HTLV-I-associated myelopathy: a multicenter, randomized, double-blind, controlled trial. Neurology. 1996, 46: 1016-1021.View ArticlePubMedGoogle Scholar
- Saito M, Nakagawa M, Kaseda S, Matsuzaki T, Jonosono M, Eiraku N, Kubota R, Takenouchi N, Nagai M, Furukawa Y, Usuku K, Izumo S, Osame M: Decreased human T lymphotropic virus type I (HTLV-I) provirus load and alteration in T cell phenotype after interferon-alpha therapy for HTLV-I-associated myelopathy/tropical spastic paraparesis. J Infect Dis. 2004, 189: 29-40. 10.1086/380101.View ArticlePubMedGoogle Scholar
- Matsuzaki T, Saito M, Usuku K, Nose H, Izumo S, Arimura K, Osame M: A prospective uncontrolled trial of fermented milk drink containing viable Lactobacillus casei strain Shirota in the treatment of HTLV-1 associated myelopathy/tropical spastic paraparesis. J Neurol Sci. 2005, 15:237 (1–2): 1-2.Google Scholar
- Kurtzke JF: Rating neurologic impairment in multiple sclerosis: an expanded disability status scale (EDSS). Neurology. 1983, 33: 1444-1452.View ArticlePubMedGoogle Scholar
- Kamihira S, Sugahara K, Tsuruda K, Minami S, Uemura A, Akamatsu N, Nagai H, Murata K, Hasegawa H, Hirakata Y, Takasaki Y, Tsukasaki K, Yamada Y: Proviral status of HTLV-1 integrated into the host genomic DNA of adult T-cell leukemia cells. Clin Lab Haematol. 2005, 27: 235-241. 10.1111/j.1365-2257.2005.00698.x.View ArticlePubMedGoogle Scholar
- Hasegawa A, Ohashi T, Hanabuchi S, Kato H, Takemura F, Masuda T, Kannagi M: Expansion of human T-cell leukemia virus type 1 (HTLV-1) reservoir in orally infected rats: inverse correlation with HTLV-1-specific cellular immune response. J Virol. 2003, 77: 2956-2963. 10.1128/JVI.77.5.2956-2963.2003.PubMed CentralView ArticlePubMedGoogle Scholar
- Shimizu Y, Takamori A, Utsunomiya A, Kurimura M, Yamano Y, Hishizawa M, Hasegawa A, Kondo F, Kurihara K, Harashima N, Watanabe T, Okamura J, Masuda T, Kannagi M: Impaired Tax-specific T-cell responses with insufficient control of HTLV-1 in a subgroup of individuals at asymptomatic and smoldering stages. Cancer Sci. 2008 in press.Google Scholar
- Lemasson I, Lewis MR, Polakowski N, Hivin P, Cavanagh MH, Thebault S, Barbeau B, Nyborg JK, Mesnard JM: Human T-cell leukemia virus type 1 (HTLV-1) bZIP protein interacts with the cellular transcription factor CREB to inhibit HTLV-1 transcription. J Virol. 2007, 81: 1543-1553. 10.1128/JVI.00480-06.PubMed CentralView ArticlePubMedGoogle Scholar
- Fuchs D, Hausen A, Reibnegger G, Werner ER, Dierich MP, Wachter H: Neopterin as a marker for activated cell-mediated immunity: application in HIV infection. Immunol Today. 1988, 9: 150-155. 10.1016/0167-5699(88)91203-0.View ArticlePubMedGoogle Scholar
- Furukawa Y, Kubota R, Eiraku N, Nakagawa M, Usuku K, Izumo S, Osame M: Human T-cell lymphotropic virus type I (HTLV-I)-related clinical and laboratory findings for HTLV-I-infected blood donors. J Acquir Immune Defic Syndr. 2003, 32: 328-334. 10.1097/00126334-200303010-00013.View ArticlePubMedGoogle Scholar
- Jeffery KJ, Usuku K, Hall SE, Matsumoto W, Taylor GP, Procter J, Bunce M, Ogg GS, Welsh KI, Weber JN, Lloyd AL, Nowak MA, Nagai M, Kodama D, Izumo S, Osame M, Bangham CR: HLA alleles determine human T-lymphotropic virus-I (HTLV-I) proviral load and the risk of HTLV-I-associated myelopathy. Proc Natl Acad Sci USA. 1999, 96: 3848-3853. 10.1073/pnas.96.7.3848.PubMed CentralView ArticlePubMedGoogle Scholar
- Jeffery KJ, Siddiqui AA, Bunce M, Lloyd AL, Vine AM, Witkover AD, Izumo S, Usuku K, Welsh KI, Osame M, Bangham CR: The influence of HLA class I alleles and heterozygosity on the outcome of human T cell lymphotropic virus type I infection. J Immunol. 2000, 165: 7278-7284.View ArticlePubMedGoogle Scholar
- Sabouri AH, Saito M, Usuku K, Bajestan SN, Mahmoudi M, Forughipour M, Sabouri Z, Abbaspour Z, Goharjoo ME, Khayami E, Hasani A, Izumo S, Arimura K, Farid R, Osame M: Differences in viral and host genetic risk factors for development of human T-cell lymphotropic virus type 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis between Iranian and Japanese HTLV-1-infected individuals. J Gen Virol. 2005, 86: 773-781. 10.1099/vir.0.80509-0.View ArticlePubMedGoogle Scholar
- Furukawa Y, Yamashita M, Usuku K, Izumo S, Nakagawa M, Osame M: 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. 2000, 182: 1343-1349. 10.1086/315897.View ArticlePubMedGoogle Scholar
- Birmann BM, Breen EC, Stuver S, Cranston B, Martinez-Maza O, Falk KI, Okayama A, Hanchard B, Mueller N, Hisada M: Population differences in immune marker profiles associated with human T-lymphotropic virus type I infection in Japan and Jamaica. Int J Cancer. 2008, 1:124 (3): 614-21.Google Scholar
- Nakagawa M, Nakahara K, Maruyama Y, Kawabata M, Higuchi I, Kubota H, Izumo S, Arimura K, Osame M: Therapeutic trials in 200 patients with HTLV-I-associated myelopathy/tropical spastic paraparesis. J Neurovirol. 1996, 2: 345-355. 10.3109/13550289609146899.View ArticlePubMedGoogle Scholar
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