Abstract
We transfected human and rabbit peripheral blood mononuclear cells (PBMC) with the ACH molecular clone of human T-cell lymphotropic virus type 1 (HTLV-1) to study its in vitro and in vivo properties. PBMC transfected with ACH were shown to transfer infection to naive PBMC. ACH transformed rabbit PBMC, as indicated by interleukin-2-independent proliferation of a transfectant culture. This transformant culture was shown by flow cytometric analysis to be a CD4+ CD25+ T-lymphocyte population containing, as determined by Southern blot analysis, at least three integrated HTLV-1 proviral copies. HTLV-1 infection was produced in rabbits inoculated with ACH-transfected, irradiated PBMC. Inoculated rabbits seroconverted to positivity for antibodies against HTLV-1 and had steady or rising HTLV-1 enzyme-linked immunosorbent assay antibody titers. Western blot (immunoblot) analysis revealed sustained seroconversion of rabbits to positivity for antibodies against all major viral antigenic determinants. Infection of rabbits was further demonstrated by antigen capture assay of p24 in PBMC and lymph node cultures and PCR amplification of proviral sequences from PBMC. These data suggest that ACH, like wild-type HTLV-1, infects and transforms primary CD4+ T lymphocytes and is infectious in vivo. This clone will facilitate investigations into the role of viral genes on biological properties of HTLV-1 in vitro and in vivo.
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- Akagi T., Takeda I., Oka T., Ohtsuki Y., Yano S., Miyoshi I. Experimental infection of rabbits with human T-cell leukemia virus type I. Jpn J Cancer Res. 1985 Feb;76(2):86–94. [PubMed] [Google Scholar]
- Baum C., Forster P., Hegewisch-Becker S., Harbers K. An optimized electroporation protocol applicable to a wide range of cell lines. Biotechniques. 1994 Dec;17(6):1058–1062. [PubMed] [Google Scholar]
- Brown D. W., Blais B. P., Robinson H. L. Long terminal repeat (LTR) sequences, env, and a region near the 5' LTR influence the pathogenic potential of recombinants between Rous-associated virus types 0 and 1. J Virol. 1988 Sep;62(9):3431–3437. doi: 10.1128/jvi.62.9.3431-3437.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cockerell G. L., Lairmore M., De B., Rovnak J., Hartley T. M., Miyoshi I. Persistent infection of rabbits with HTLV-I: patterns of anti-viral antibody reactivity and detection of virus by gene amplification. Int J Cancer. 1990 Jan 15;45(1):127–130. doi: 10.1002/ijc.2910450123. [DOI] [PubMed] [Google Scholar]
- Derse D., Mikovits J., Polianova M., Felber B. K., Ruscetti F. Virions released from cells transfected with a molecular clone of human T-cell leukemia virus type I give rise to primary and secondary infections of T cells. J Virol. 1995 Mar;69(3):1907–1912. doi: 10.1128/jvi.69.3.1907-1912.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dobrescu D., Ursea B., Pope M., Asch A. S., Posnett D. N. Enhanced HIV-1 replication in V beta 12 T cells due to human cytomegalovirus in monocytes: evidence for a putative herpesvirus superantigen. Cell. 1995 Sep 8;82(5):753–763. doi: 10.1016/0092-8674(95)90472-7. [DOI] [PubMed] [Google Scholar]
- Donahue P. R., Quackenbush S. L., Gallo M. V., deNoronha C. M., Overbaugh J., Hoover E. A., Mullins J. I. Viral genetic determinants of T-cell killing and immunodeficiency disease induction by the feline leukemia virus FeLV-FAIDS. J Virol. 1991 Aug;65(8):4461–4469. doi: 10.1128/jvi.65.8.4461-4469.1991. [DOI] [PMC free article] [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]
- Fisher A. G., Collalti E., Ratner L., Gallo R. C., Wong-Staal F. A molecular clone of HTLV-III with biological activity. Nature. 1985 Jul 18;316(6025):262–265. doi: 10.1038/316262a0. [DOI] [PubMed] [Google Scholar]
- Franchini G. Molecular mechanisms of human T-cell leukemia/lymphotropic virus type I infection. Blood. 1995 Nov 15;86(10):3619–3639. [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]
- Hartley T. M., Khabbaz R. F., Cannon R. O., Kaplan J. E., Lairmore M. D. Characterization of antibody reactivity to human T-cell lymphotropic virus types I and II using immunoblot and radioimmunoprecipitation assays. J Clin Microbiol. 1990 Apr;28(4):646–650. doi: 10.1128/jcm.28.4.646-650.1990. [DOI] [PMC free article] [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]
- Herrmann B. G., Frischauf A. M. Isolation of genomic DNA. Methods Enzymol. 1987;152:180–183. doi: 10.1016/0076-6879(87)52018-3. [DOI] [PubMed] [Google Scholar]
- Itoyama Y., Minato S., Kira J., Goto I., Sato H., Okochi K., Yamamoto N. Spontaneous proliferation of peripheral blood lymphocytes increased in patients with HTLV-I-associated myelopathy. Neurology. 1988 Aug;38(8):1302–1307. doi: 10.1212/wnl.38.8.1302. [DOI] [PubMed] [Google Scholar]
- Kimata J. T., Ratner L. Temporal regulation of viral and cellular gene expression during human T-lymphotropic virus type I-mediated lymphocyte immortalization. J Virol. 1991 Aug;65(8):4398–4407. doi: 10.1128/jvi.65.8.4398-4407.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kimata J. T., Wong F. H., Wang J. J., Ratner L. Construction and characterization of infectious human T-cell leukemia virus type 1 molecular clones. Virology. 1994 Nov 1;204(2):656–664. doi: 10.1006/viro.1994.1581. [DOI] [PubMed] [Google Scholar]
- Korber B., Okayama A., Donnelly R., Tachibana N., Essex M. Polymerase chain reaction analysis of defective human T-cell leukemia virus type I proviral genomes in leukemic cells of patients with adult T-cell leukemia. J Virol. 1991 Oct;65(10):5471–5476. doi: 10.1128/jvi.65.10.5471-5476.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lairmore M. D., DiGeorge A. M., Conrad S. F., Trevino A. V., Lal R. B., Kaumaya P. T. Human T-lymphotropic virus type 1 peptides in chimeric and multivalent constructs with promiscuous T-cell epitopes enhance immunogenicity and overcome genetic restriction. J Virol. 1995 Oct;69(10):6077–6089. doi: 10.1128/jvi.69.10.6077-6089.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lairmore M. D., Roberts B., Frank D., Rovnak J., Weiser M. G., Cockerell G. L. Comparative biological responses of rabbits infected with human T-lymphotropic virus type I isolates from patients with lymphoproliferative and neurodegenerative disease. Int J Cancer. 1992 Jan 2;50(1):124–130. doi: 10.1002/ijc.2910500125. [DOI] [PubMed] [Google Scholar]
- Luciw P. A., Shaw K. E., Unger R. E., Planelles V., Stout M. W., Lackner J. E., Pratt-Lowe E., Leung N. J., Banapour B., Marthas M. L. Genetic and biological comparisons of pathogenic and nonpathogenic molecular clones of simian immunodeficiency virus (SIVmac). AIDS Res Hum Retroviruses. 1992 Mar;8(3):395–402. doi: 10.1089/aid.1992.8.395. [DOI] [PubMed] [Google Scholar]
- Matsumoto M., Sugimoto M., Nakashima H., Imamura F., Kawano O., Uyama E., Takatsu K., Araki S. Spontaneous T cell proliferation and release of soluble interleukin-2 receptors in patients with HTLV-I-associated myelopathy. Am J Trop Med Hyg. 1990 Apr;42(4):365–373. doi: 10.4269/ajtmh.1990.42.365. [DOI] [PubMed] [Google Scholar]
- Merl S., Kloster B., Moore J., Hubbell C., Tomar R., Davey F., Kalinowski D., Planas A., Ehrlich G., Clark D. Efficient transformation of previously activated and dividing T lymphocytes by human T cell leukemia-lymphoma virus. Blood. 1984 Nov;64(5):967–974. [PubMed] [Google 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 Mar;83(3):236–239. doi: 10.1111/j.1349-7006.1992.tb00092.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgan O. S., Rodgers-Johnson P., Mora C., Char G. HTLV-1 and polymyositis in Jamaica. Lancet. 1989 Nov 18;2(8673):1184–1187. doi: 10.1016/s0140-6736(89)91793-5. [DOI] [PubMed] [Google Scholar]
- Newbound G. C., Andrews J. M., O'Rourke J. P., Brady J. N., Lairmore M. D. Human T-cell lymphotropic virus type 1 Tax mediates enhanced transcription in CD4+ T lymphocytes. J Virol. 1996 Apr;70(4):2101–2106. doi: 10.1128/jvi.70.4.2101-2106.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nishioka K., Maruyama I., Sato K., Kitajima I., Nakajima Y., Osame M. Chronic inflammatory arthropathy associated with HTLV-I. Lancet. 1989 Feb 25;1(8635):441–441. doi: 10.1016/s0140-6736(89)90038-x. [DOI] [PubMed] [Google Scholar]
- Ohshima K., Kikuchi M., Masuda Y., Kobari S., Sumiyoshi Y., Eguchi F., Mohtai H., Yoshida T., Takeshita M., Kimura N. Defective provirus form of human T-cell leukemia virus type I in adult T-cell leukemia/lymphoma: clinicopathological features. Cancer Res. 1991 Sep 1;51(17):4639–4642. [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]
- Persaud D., Muñoz J. L., Tarsis S. L., Parks E. S., Parks W. P. Time course and cytokine dependence of human T-cell lymphotropic virus type 1 T-lymphocyte transformation as revealed by a microtiter infectivity assay. J Virol. 1995 Oct;69(10):6297–6303. doi: 10.1128/jvi.69.10.6297-6303.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Prince H. E., Golding J., York J. Lymphocyte subset alterations associated with increased spontaneous lymphocyte proliferation in human T lymphotropic virus (HTLV) infection: distinctive patterns for HTLV-I versus HTLV-II infection. J Infect Dis. 1994 Jun;169(6):1409–1410. doi: 10.1093/infdis/169.6.1409. [DOI] [PubMed] [Google Scholar]
- Prince H. E., Lee H., Jensen E. R., Swanson P., Weber D., Fitzpatrick L., Doyle M., Kleinman S. Immunologic correlates of spontaneous lymphocyte proliferation in human T-lymphotropic virus infection. Blood. 1991 Jul 1;78(1):169–174. [PubMed] [Google Scholar]
- Robinson H. L., Blais B. M., Tsichlis P. N., Coffin J. M. At least two regions of the viral genome determine the oncogenic potential of avian leukosis viruses. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1225–1229. doi: 10.1073/pnas.79.4.1225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rowe T., Dezzutti C., Guenthner P. C., Lam L., Hodge T., Lairmore M. D., Lal R. B., Folks T. M. Characterization of a HTLV-I-infected cell line derived from a patient with adult T-cell leukemia with stable co-expression of CD4 and CD8. Leuk Res. 1995 Sep;19(9):621–628. doi: 10.1016/0145-2126(95)00030-r. [DOI] [PubMed] [Google Scholar]
- Tokudome S., Tokunaga O., Shimamoto Y., Miyamoto Y., Sumida I., Kikuchi M., Takeshita M., Ikeda T., Fujiwara K., Yoshihara M. Incidence of adult T-cell leukemia/lymphoma among human T-lymphotropic virus type I carriers in Saga, Japan. Cancer Res. 1989 Jan 1;49(1):226–228. [PubMed] [Google Scholar]
- Willems L., Kerkhofs P., Dequiedt F., Portetelle D., Mammerickx M., Burny A., Kettmann R. Attenuation of bovine leukemia virus by deletion of R3 and G4 open reading frames. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11532–11536. doi: 10.1073/pnas.91.24.11532. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamaguchi K., Takatsuki K. Adult T cell leukaemia-lymphoma. Baillieres Clin Haematol. 1993 Dec;6(4):899–915. doi: 10.1016/s0950-3536(05)80183-0. [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 T. M., Robinson M. A., Bowers F. S., Kindt T. J. Characterization of an infectious molecular clone of human T-cell leukemia virus type I. J Virol. 1995 Apr;69(4):2024–2030. doi: 10.1128/jvi.69.4.2024-2030.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]