Abstract
The ability of molecular clones of human T-cell leukemia virus type I (HTLV-I) to direct the synthesis of infectious virions has not previously been demonstrated. An HTLV-I provirus originating from an adult T-cell leukemia patient was cloned into a plasmid vector and is designated pCS-HTLV. This molecular clone was shown to direct the synthesis of viral mRNA and proteins in transiently transfected cells; in addition, virus structural proteins were released into the culture medium. Viral proteins were assembled into virions that sedimented at a buoyant density characteristic of retrovirus particles and whose morphology was verified by electron microscopy. Virions concentrated from transiently transfected cell supernatants were incubated with primary cord blood lymphocytes or with transformed T-cell lines to establish that these particles were infectious. Expression of spliced, viral mRNAs in the T-cell cultures after both primary and secondary infections with cell-free virus revealed that pCS-HTLV encodes an infectious provirus.
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- Ciminale V., Pavlakis G. N., Derse D., Cunningham C. P., Felber B. K. Complex splicing in the human T-cell leukemia virus (HTLV) family of retroviruses: novel mRNAs and proteins produced by HTLV type I. J Virol. 1992 Mar;66(3):1737–1745. doi: 10.1128/jvi.66.3.1737-1745.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clapham P., Nagy K., Cheingsong-Popov R., Exley M., Weiss R. A. Productive infection and cell-free transmission of human T-cell leukemia virus in a nonlymphoid cell line. Science. 1983 Dec 9;222(4628):1125–1127. doi: 10.1126/science.6316502. [DOI] [PubMed] [Google Scholar]
- Ehrlich G. D., Davey F. R., Kirshner J. J., Sninsky J. J., Kwok S., Slamon D. J., Kalish R., Poiesz B. J. A polyclonal CD4+ and CD8+ lymphocytosis in a patient doubly infected with HTLV-I and HIV-1: a clinical and molecular analysis. Am J Hematol. 1989 Mar;30(3):128–139. doi: 10.1002/ajh.2830300304. [DOI] [PubMed] [Google Scholar]
- Fan N., Gavalchin J., Paul B., Wells K. H., Lane M. J., Poiesz B. J. Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I. J Clin Microbiol. 1992 Apr;30(4):905–910. doi: 10.1128/jcm.30.4.905-910.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Felber B. K., Derse D., Athanassopoulos A., Campbell M., Pavlakis G. N. Cross-activation of the Rex proteins of HTLV-I and BLV and of the Rev protein of HIV-1 and nonreciprocal interactions with their RNA responsive elements. New Biol. 1989 Dec;1(3):318–328. [PubMed] [Google Scholar]
- Felber B. K., Paskalis H., Kleinman-Ewing C., Wong-Staal F., Pavlakis G. N. The pX protein of HTLV-I is a transcriptional activator of its long terminal repeats. Science. 1985 Aug 16;229(4714):675–679. doi: 10.1126/science.2992082. [DOI] [PubMed] [Google Scholar]
- Fujisawa J., Seiki M., Kiyokawa T., Yoshida M. Functional activation of the long terminal repeat of human T-cell leukemia virus type I by a trans-acting factor. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2277–2281. doi: 10.1073/pnas.82.8.2277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gendelman H. E., Friedman R. M., Joe S., Baca L. M., Turpin J. A., Dveksler G., Meltzer M. S., Dieffenbach C. A selective defect of interferon alpha production in human immunodeficiency virus-infected monocytes. J Exp Med. 1990 Nov 1;172(5):1433–1442. doi: 10.1084/jem.172.5.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gessain A., Barin F., Vernant J. C., Gout O., Maurs L., Calender A., de Thé G. Antibodies to human T-lymphotropic virus type-I in patients with tropical spastic paraparesis. Lancet. 1985 Aug 24;2(8452):407–410. doi: 10.1016/s0140-6736(85)92734-5. [DOI] [PubMed] [Google Scholar]
- Gessain A., Saal F., Giron M. L., Lasneret J., Lagaye S., Gout O., De Thé G., Sigaux F., Peries J. Cell surface phenotype and human T lymphotropic virus type 1 antigen expression in 12 T cell lines derived from peripheral blood and cerebrospinal fluid of West Indian, Guyanese and African patients with tropical spastic paraparesis. J Gen Virol. 1990 Feb;71(Pt 2):333–341. doi: 10.1099/0022-1317-71-2-333. [DOI] [PubMed] [Google Scholar]
- Hattori T., Uchiyama T., Toibana T., Takatsuki K., Uchino H. Surface phenotype of Japanese adult T-cell leukemia cells characterized by monoclonal antibodies. Blood. 1981 Sep;58(3):645–647. [PubMed] [Google Scholar]
- Hinuma Y., Nagata K., Hanaoka M., Nakai M., Matsumoto T., Kinoshita K. I., 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 U S A. 1981 Oct;78(10):6476–6480. doi: 10.1073/pnas.78.10.6476. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hiramatsu K., Masuda M., Yoshikura H. Mode of transmission of human T-cell leukemia virus type I (HTLV I) in a human promyelocytic leukemia HL60 cell. Int J Cancer. 1986 Apr 15;37(4):601–606. doi: 10.1002/ijc.2910370420. [DOI] [PubMed] [Google Scholar]
- Ho D. D., Rota T. R., Hirsch M. S. Infection of human endothelial cells by human T-lymphotropic virus type I. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7588–7590. doi: 10.1073/pnas.81.23.7588. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoffman P. M., Dhib-Jalbut S., Mikovits J. A., Robbins D. S., Wolf A. L., Bergey G. K., Lohrey N. C., Weislow O. S., Ruscetti F. W. Human T-cell leukemia virus type I infection of monocytes and microglial cells in primary human cultures. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11784–11788. doi: 10.1073/pnas.89.24.11784. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoxie J. A., Matthews D. M., Cines D. B. Infection of human endothelial cells by human T-cell leukemia virus type I. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7591–7595. doi: 10.1073/pnas.81.23.7591. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inoue J., Seiki M., Yoshida M. The second pX product p27 chi-III of HTLV-1 is required for gag gene expression. FEBS Lett. 1986 Dec 15;209(2):187–190. doi: 10.1016/0014-5793(86)81108-5. [DOI] [PubMed] [Google Scholar]
- Inoue J., Yoshida M., Seiki M. Transcriptional (p40x) and post-transcriptional (p27x-III) regulators are required for the expression and replication of human T-cell leukemia virus type I genes. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3653–3657. doi: 10.1073/pnas.84.11.3653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Longo D. L., Gelmann E. P., Cossman J., Young R. A., Gallo R. C., O'Brien S. J., Matis L. A. Isolation of HTLV-transformed B-lymphocyte clone from a patient with HTLV-associated adult T-cell leukaemia. Nature. 1984 Aug 9;310(5977):505–506. doi: 10.1038/310505a0. [DOI] [PubMed] [Google Scholar]
- Mann D. L., Clark J., Clarke M., Reitz M., Popovic M., Franchini G., Trainor C. D., Strong D. M., Blattner W. A., Gallo R. C. Identification of the human T cell lymphoma virus in B cell lines established from patients with adult T cell leukemia. J Clin Invest. 1984 Jul;74(1):56–62. doi: 10.1172/JCI111418. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martarano L., Stephens R., Rice N., Derse D. Equine infectious anemia virus trans-regulatory protein Rev controls viral mRNA stability, accumulation, and alternative splicing. J Virol. 1994 May;68(5):3102–3111. doi: 10.1128/jvi.68.5.3102-3111.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mori K., Sabe H., Siomi H., Iino T., Tanaka A., Takeuchi K., Hirayoshi K., Hatanaka M. Expression of a provirus of human T cell leukaemia virus type I by DNA transfection. J Gen Virol. 1987 Feb;68(Pt 2):499–506. doi: 10.1099/0022-1317-68-2-499. [DOI] [PubMed] [Google Scholar]
- Nicot C., Astier-Gin T., Edouard E., Legrand E., Moynet D., Vital A., Londos-Gagliardi D., Moreau J. P., Guillemain B. Establishment of HTLV-I-infected cell lines from French, Guianese and West Indian patients and isolation of a proviral clone producing viral particles. Virus Res. 1993 Dec;30(3):317–334. doi: 10.1016/0168-1702(93)90099-9. [DOI] [PubMed] [Google Scholar]
- Okada M., Koyanagi Y., Kobayashi N., Tanaka Y., Nakai M., Sano K., Takeuchi K., Hinuma Y., Hatanaka M., Yamamoto N. In vitro infection of human B lymphocytes with adult T-cell leukemia virus. Cancer Lett. 1984 Feb;22(1):11–21. doi: 10.1016/0304-3835(84)90038-7. [DOI] [PubMed] [Google 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 May 3;1(8488):1031–1032. doi: 10.1016/s0140-6736(86)91298-5. [DOI] [PubMed] [Google Scholar]
- Poiesz B. J., Ruscetti F. W., Gazdar A. F., Bunn P. A., Minna J. D., Gallo R. C. 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. 1980 Dec;77(12):7415–7419. doi: 10.1073/pnas.77.12.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poiesz B. J., Ruscetti F. W., Reitz M. S., Kalyanaraman V. S., Gallo R. C. Isolation of a new type C retrovirus (HTLV) in primary uncultured cells of a patient with Sézary T-cell leukaemia. Nature. 1981 Nov 19;294(5838):268–271. doi: 10.1038/294268a0. [DOI] [PubMed] [Google Scholar]
- Richardson J. H., Edwards A. J., Cruickshank J. K., Rudge P., Dalgleish A. G. In vivo cellular tropism of human T-cell leukemia virus type 1. J Virol. 1990 Nov;64(11):5682–5687. doi: 10.1128/jvi.64.11.5682-5687.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruscetti F. W., Mikovits J. A., Kalyanaraman V. S., Overton R., Stevenson H., Stromberg K., Herberman R. B., Farrar W. L., Ortaldo J. R. Analysis of effector mechanisms against HTLV-I- and HTLV-III/LAV-infected lymphoid cells. J Immunol. 1986 May 15;136(10):3619–3624. [PubMed] [Google Scholar]
- Ruscetti F. W., Robert-Guroff M., Ceccherini-Nelli L., Minowada J., Popovic M., Gallo R. C. Persistent in vitro infection by human T-cell leukemia-lymphoma virus (HTLV) of normal human T-lymphocytes from blood relatives of patients with HTLV-associated mature T-cell neoplasms. Int J Cancer. 1983 Feb 15;31(2):171–180. doi: 10.1002/ijc.2910310207. [DOI] [PubMed] [Google Scholar]
- Saida T., Saida K., Funauchi M., Nishiguchi E., Nakajima M., Matsuda S., Ohta M., Ohta K., Nishitani H., Hatanaka M. HTLV-I myelitis: isolation of virus, genomic analysis, and infection in neural cell cultures. Ann N Y Acad Sci. 1988;540:636–638. doi: 10.1111/j.1749-6632.1988.tb27196.x. [DOI] [PubMed] [Google Scholar]
- Saksela K., Muchmore E., Girard M., Fultz P., Baltimore D. High viral load in lymph nodes and latent human immunodeficiency virus (HIV) in peripheral blood cells of HIV-1-infected chimpanzees. J Virol. 1993 Dec;67(12):7423–7427. doi: 10.1128/jvi.67.12.7423-7427.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seiki M., Inoue J., Takeda T., Yoshida M. Direct evidence that p40x of human T-cell leukemia virus type I is a trans-acting transcriptional activator. EMBO J. 1986 Mar;5(3):561–565. doi: 10.1002/j.1460-2075.1986.tb04247.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sodroski J. G., Rosen C. A., Haseltine W. A. Trans-acting transcriptional activation of the long terminal repeat of human T lymphotropic viruses in infected cells. Science. 1984 Jul 27;225(4660):381–385. doi: 10.1126/science.6330891. [DOI] [PubMed] [Google Scholar]
- Sodroski J. The human T-cell leukemia virus (HTLV) transactivator (Tax) protein. Biochim Biophys Acta. 1992 Sep 14;1114(1):19–29. doi: 10.1016/0304-419x(92)90003-h. [DOI] [PubMed] [Google Scholar]
- Sugamura K., Sakitani M., Hinuma Y. Microplate method for retrovirus-induced transformation of normal human T-cells. J Immunol Methods. 1984 Oct 26;73(2):379–385. doi: 10.1016/0022-1759(84)90413-7. [DOI] [PubMed] [Google Scholar]
- Yamamoto N., Matsumoto T., Koyanagi Y., Tanaka Y., Hinuma Y. Unique cell lines harbouring both Epstein-Barr virus and adult T-cell leukaemia virus, established from leukaemia patients. Nature. 1982 Sep 23;299(5881):367–369. doi: 10.1038/299367a0. [DOI] [PubMed] [Google 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 U S A. 1982 Mar;79(6):2031–2035. doi: 10.1073/pnas.79.6.2031. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhao L. J., Giam C. Z. Interaction of the human T-cell lymphotrophic virus type I (HTLV-I) transcriptional activator Tax with cellular factors that bind specifically to the 21-base-pair repeats in the HTLV-I enhancer. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11445–11449. doi: 10.1073/pnas.88.24.11445. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Rossi A., Aldovini A., Franchini G., Mann D., Gallo R. C., Wong-Staal F. Clonal selection of T lymphocytes infected by cell-free human T-cell leukemia/lymphoma virus type I: parameters of virus integration and expression. Virology. 1985 Jun;143(2):640–645. doi: 10.1016/0042-6822(85)90405-2. [DOI] [PubMed] [Google Scholar]