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J Virol. Apr 1995; 69(4): 2159–2167.
PMCID: PMC188884

Restriction of human immunodeficiency virus type 1 production in a human astrocytoma cell line is associated with a cellular block in Rev function.


Chronically human immunodeficiency virus type 1 (HIV-1) strain IIIB-infected human TH4-7-5 astrocytoma cells show low-level virus production. Cocultivation of TH4-7-5 cells with myelomonocytic cells led to active virus production in these target cells after a lag period, indicating cell-determined restriction of virus replication in the glial cells. HIV-1 transcript patterns of TH4-7-5 cells contained only a small proportion of Rev-dependent mRNA species, mimicking a Rev-negative phenotype despite the presence of rev mRNAs and protein. Sequencing of the single provirus integrated in TH4-7-5 cells demonstrated that the rev gene and the Rev-responsive element are intact. These results suggested inhibited function of the Rev-regulatory unit in these astrocytoma cells. Transfection of TH4-7-5 cells with a Rev expression plasmid resulted in weak or no induction of proviral p24gag antigen levels compared with the dramatic increase observed in Rev-permissive HeLa cells. Immunofluorescence analysis of TH4-7-5 cells transfected with a rev-expressing plasmid revealed prominent cytoplasmic and nuclear-nucleolar localization of Rev, in contrast to the predominant nuclear-nucleolar localization pattern of Rev in HeLa cells. We conclude that restriction of virus production in TH4-7-5 cells is at least partially due to a block in Rev-dependent posttranscriptional regulation of HIV expression.

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Selected References

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  • Ahmed YF, Hanly SM, Malim MH, Cullen BR, Greene WC. Structure-function analyses of the HTLV-I Rex and HIV-1 Rev RNA response elements: insights into the mechanism of Rex and Rev action. Genes Dev. 1990 Jun;4(6):1014–1022. [PubMed]
  • Arrigo SJ, Chen IS. Rev is necessary for translation but not cytoplasmic accumulation of HIV-1 vif, vpr, and env/vpu 2 RNAs. Genes Dev. 1991 May;5(5):808–819. [PubMed]
  • Atwood WJ, Berger JR, Kaderman R, Tornatore CS, Major EO. Human immunodeficiency virus type 1 infection of the brain. Clin Microbiol Rev. 1993 Oct;6(4):339–366. [PMC free article] [PubMed]
  • Benko DM, Robinson R, Solomin L, Mellini M, Felber BK, Pavlakis GN. Binding of trans-dominant mutant Rev protein of human immunodeficiency virus type 1 to the cis-acting Rev-responsive element does not affect the fate of viral mRNA. New Biol. 1990 Dec;2(12):1111–1122. [PubMed]
  • Bigner DD, Bigner SH, Pontén J, Westermark B, Mahaley MS, Ruoslahti E, Herschman H, Eng LF, Wikstrand CJ. Heterogeneity of Genotypic and phenotypic characteristics of fifteen permanent cell lines derived from human gliomas. J Neuropathol Exp Neurol. 1981 May;40(3):201–229. [PubMed]
  • Brack-Werner R, Kleinschmidt A, Ludvigsen A, Mellert W, Neumann M, Herrmann R, Khim MC, Burny A, Müller-Lantzsch N, Stavrou D, et al. Infection of human brain cells by HIV-1: restricted virus production in chronically infected human glial cell lines. AIDS. 1992 Mar;6(3):273–285. [PubMed]
  • Bradley TR, Pilkington G, Garson M, Hodgson GS, Kraft N. Cell lines derived from a human myelomonocytic leukaemia. Br J Haematol. 1982 Aug;51(4):595–604. [PubMed]
  • Cannon P, Kim SH, Ulich C, Kim S. Analysis of Tat function in human immunodeficiency virus type 1-infected low-level-expression cell lines U1 and ACH-2. J Virol. 1994 Mar;68(3):1993–1997. [PMC free article] [PubMed]
  • Chatterjee B, Gimbel W, Görblich T, Werner T. HPLC purification of PCR products for direct sequencing. Trends Genet. 1993 Dec;9(12):406–406. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Cullen BR. Human immunodeficiency virus as a prototypic complex retrovirus. J Virol. 1991 Mar;65(3):1053–1056. [PMC free article] [PubMed]
  • D'Agostino DM, Felber BK, Harrison JE, Pavlakis GN. The Rev protein of human immunodeficiency virus type 1 promotes polysomal association and translation of gag/pol and vpu/env mRNAs. Mol Cell Biol. 1992 Mar;12(3):1375–1386. [PMC free article] [PubMed]
  • Dayton ET, Powell DM, Dayton AI. Functional analysis of CAR, the target sequence for the Rev protein of HIV-1. Science. 1989 Dec 22;246(4937):1625–1629. [PubMed]
  • de Wet JR, Wood KV, DeLuca M, Helinski DR, Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. [PMC free article] [PubMed]
  • Emerman M, Vazeux R, Peden K. The rev gene product of the human immunodeficiency virus affects envelope-specific RNA localization. Cell. 1989 Jun 30;57(7):1155–1165. [PubMed]
  • Fankhauser C, Izaurralde E, Adachi Y, Wingfield P, Laemmli UK. Specific complex of human immunodeficiency virus type 1 rev and nucleolar B23 proteins: dissociation by the Rev response element. Mol Cell Biol. 1991 May;11(5):2567–2575. [PMC free article] [PubMed]
  • Feinberg MB, Jarrett RF, Aldovini A, Gallo RC, Wong-Staal F. HTLV-III expression and production involve complex regulation at the levels of splicing and translation of viral RNA. Cell. 1986 Sep 12;46(6):807–817. [PubMed]
  • Felber BK, Drysdale CM, Pavlakis GN. Feedback regulation of human immunodeficiency virus type 1 expression by the Rev protein. J Virol. 1990 Aug;64(8):3734–3741. [PMC free article] [PubMed]
  • Felber BK, Hadzopoulou-Cladaras M, Cladaras C, Copeland T, Pavlakis GN. rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1495–1499. [PMC free article] [PubMed]
  • Felber BK, Pavlakis GN. Molecular biology of HIV-1: positive and negative regulatory elements important for virus expression. AIDS. 1993;7 (Suppl 1):S51–S62. [PubMed]
  • Folks TM, Clouse KA, Justement J, Rabson A, Duh E, Kehrl JH, Fauci AS. Tumor necrosis factor alpha induces expression of human immunodeficiency virus in a chronically infected T-cell clone. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2365–2368. [PMC free article] [PubMed]
  • Folks TM, Justement J, Kinter A, Dinarello CA, Fauci AS. Cytokine-induced expression of HIV-1 in a chronically infected promonocyte cell line. Science. 1987 Nov 6;238(4828):800–802. [PubMed]
  • Folks TM, Justement J, Kinter A, Schnittman S, Orenstein J, Poli G, Fauci AS. Characterization of a promonocyte clone chronically infected with HIV and inducible by 13-phorbol-12-myristate acetate. J Immunol. 1988 Feb 15;140(4):1117–1122. [PubMed]
  • Garry RF, Koch G. Tat contains a sequence related to snake neurotoxins. AIDS. 1992 Dec;6(12):1541–1542. [PubMed]
  • Geleziunas R, Schipper HM, Wainberg MA. Pathogenesis and therapy of HIV-1 infection of the central nervous system. AIDS. 1992 Dec;6(12):1411–1426. [PubMed]
  • Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. [PubMed]
  • Greene WC. Regulation of HIV-1 gene expression. Annu Rev Immunol. 1990;8:453–475. [PubMed]
  • Hadzopoulou-Cladaras M, Felber BK, Cladaras C, Athanassopoulos A, Tse A, Pavlakis GN. The rev (trs/art) protein of human immunodeficiency virus type 1 affects viral mRNA and protein expression via a cis-acting sequence in the env region. J Virol. 1989 Mar;63(3):1265–1274. [PMC free article] [PubMed]
  • Hammarskjöld ML, Heimer J, Hammarskjöld B, Sangwan I, Albert L, Rekosh D. Regulation of human immunodeficiency virus env expression by the rev gene product. J Virol. 1989 May;63(5):1959–1966. [PMC free article] [PubMed]
  • Hope TJ, Huang XJ, McDonald D, Parslow TG. Steroid-receptor fusion of the human immunodeficiency virus type 1 Rev transactivator: mapping cryptic functions of the arginine-rich motif. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7787–7791. [PMC free article] [PubMed]
  • Hope TJ, McDonald D, Huang XJ, Low J, Parslow TG. Mutational analysis of the human immunodeficiency virus type 1 Rev transactivator: essential residues near the amino terminus. J Virol. 1990 Nov;64(11):5360–5366. [PMC free article] [PubMed]
  • Kleinschmidt A, Neumann M, Möller C, Erfle V, Brack-Werner R. Restricted expression of HIV1 in human astrocytes: molecular basis for viral persistence in the CNS. Res Virol. 1994 May-Aug;145(3-4):147–153. [PubMed]
  • Kohleisen B, Neumann M, Herrmann R, Brack-Werner R, Krohn KJ, Ovod V, Ranki A, Erfle V. Cellular localization of Nef expressed in persistently HIV-1-infected low-producer astrocytes. AIDS. 1992 Dec;6(12):1427–1436. [PubMed]
  • Kubota S, Furuta R, Maki M, Hatanaka M. Inhibition of human immunodeficiency virus type 1 Rev function by a Rev mutant which interferes with nuclear/nucleolar localization of Rev. J Virol. 1992 Apr;66(4):2510–2513. [PMC free article] [PubMed]
  • Kunsch C, Wigdahl B. Transient expression of human immunodeficiency virus type 1 genome results in a nonproductive infection in human fetal dorsal root ganglia glial cells. Virology. 1989 Dec;173(2):715–722. [PubMed]
  • Laughlin MA, Zeichner S, Kolson D, Alwine JC, Seshamma T, Pomerantz RJ, Gonzalez-Scarano F. Sodium butyrate treatment of cells latently infected with HIV-1 results in the expression of unspliced viral RNA. Virology. 1993 Oct;196(2):496–505. [PubMed]
  • Lawrence JB, Cochrane AW, Johnson CV, Perkins A, Rosen CA. The HIV-1 Rev protein: a model system for coupled RNA transport and translation. New Biol. 1991 Dec;3(12):1220–1232. [PubMed]
  • Levy JA. Pathogenesis of human immunodeficiency virus infection. Microbiol Rev. 1993 Mar;57(1):183–289. [PMC free article] [PubMed]
  • Mabrouk K, Van Rietschoten J, Vives E, Darbon H, Rochat H, Sabatier JM. Lethal neurotoxicity in mice of the basic domains of HIV and SIV Rev proteins. Study of these regions by circular dichroism. FEBS Lett. 1991 Sep 2;289(1):13–17. [PubMed]
  • Malim MH, Böhnlein S, Hauber J, Cullen BR. Functional dissection of the HIV-1 Rev trans-activator--derivation of a trans-dominant repressor of Rev function. Cell. 1989 Jul 14;58(1):205–214. [PubMed]
  • Malim MH, Hauber J, Le SY, Maizel JV, Cullen BR. The HIV-1 rev trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA. Nature. 1989 Mar 16;338(6212):254–257. [PubMed]
  • Malim MH, McCarn DF, Tiley LS, Cullen BR. Mutational definition of the human immunodeficiency virus type 1 Rev activation domain. J Virol. 1991 Aug;65(8):4248–4254. [PMC free article] [PubMed]
  • Mellert W, Kleinschmidt A, Schmidt J, Festl H, Emler S, Roth WK, Erfle V. Infection of human fibroblasts and osteoblast-like cells with HIV-1. AIDS. 1990 Jun;4(6):527–535. [PubMed]
  • Mermer B, Felber BK, Campbell M, Pavlakis GN. Identification of trans-dominant HIV-1 rev protein mutants by direct transfer of bacterially produced proteins into human cells. Nucleic Acids Res. 1990 Apr 25;18(8):2037–2044. [PMC free article] [PubMed]
  • Meyer BE, Malim MH. The HIV-1 Rev trans-activator shuttles between the nucleus and the cytoplasm. Genes Dev. 1994 Jul 1;8(13):1538–1547. [PubMed]
  • Michael NL, Morrow P, Mosca J, Vahey M, Burke DS, Redfield RR. Induction of human immunodeficiency virus type 1 expression in chronically infected cells is associated primarily with a shift in RNA splicing patterns. J Virol. 1991 Mar;65(3):1291–1303. [PMC free article] [PubMed]
  • Mikovits JA, Raziuddin, Gonda M, Ruta M, Lohrey NC, Kung HF, Ruscetti FW. Negative regulation of human immune deficiency virus replication in monocytes. Distinctions between restricted and latent expression in THP-1 cells. J Exp Med. 1990 May 1;171(5):1705–1720. [PMC free article] [PubMed]
  • Nasioulas G, Zolotukhin AS, Tabernero C, Solomin L, Cunningham CP, Pavlakis GN, Felber BK. Elements distinct from human immunodeficiency virus type 1 splice sites are responsible for the Rev dependence of env mRNA. J Virol. 1994 May;68(5):2986–2993. [PMC free article] [PubMed]
  • Neumann M, Harrison J, Saltarelli M, Hadziyannis E, Erfle V, Felber BK, Pavlakis GN. Splicing variability in HIV type 1 revealed by quantitative RNA polymerase chain reaction. AIDS Res Hum Retroviruses. 1994 Nov;10(11):1531–1542. [PubMed]
  • Perkins A, Cochrane AW, Ruben SM, Rosen CA. Structural and functional characterization of the human immunodeficiency virus rev protein. J Acquir Immune Defic Syndr. 1989;2(3):256–263. [PubMed]
  • Poli G, Kinter A, Justement JS, Kehrl JH, Bressler P, Stanley S, Fauci AS. Tumor necrosis factor alpha functions in an autocrine manner in the induction of human immunodeficiency virus expression. Proc Natl Acad Sci U S A. 1990 Jan;87(2):782–785. [PMC free article] [PubMed]
  • Pomerantz RJ, Bagasra O, Baltimore D. Cellular latency of human immunodeficiency virus type 1. Curr Opin Immunol. 1992 Aug;4(4):475–480. [PubMed]
  • Pomerantz RJ, Seshamma T, Trono D. Efficient replication of human immunodeficiency virus type 1 requires a threshold level of Rev: potential implications for latency. J Virol. 1992 Mar;66(3):1809–1813. [PMC free article] [PubMed]
  • Pomerantz RJ, Trono D, Feinberg MB, Baltimore D. Cells nonproductively infected with HIV-1 exhibit an aberrant pattern of viral RNA expression: a molecular model for latency. Cell. 1990 Jun 29;61(7):1271–1276. [PubMed]
  • Popovic M, Read-Connole E, Gallo RC. T4 positive human neoplastic cell lines susceptible to and permissive for HTLV-III. Lancet. 1984 Dec 22;2(8417-8418):1472–1473. [PubMed]
  • Richard N, Iacampo S, Cochrane A. HIV-1 Rev is capable of shuttling between the nucleus and cytoplasm. Virology. 1994 Oct;204(1):123–131. [PubMed]
  • Rosen CA, Pavlakis GN. Tat and Rev: positive regulators of HIV gene expression. AIDS. 1990 Jun;4(6):499–509. [PubMed]
  • Sabatier JM, Vives E, Mabrouk K, Benjouad A, Rochat H, Duval A, Hue B, Bahraoui E. Evidence for neurotoxic activity of tat from human immunodeficiency virus type 1. J Virol. 1991 Feb;65(2):961–967. [PMC free article] [PubMed]
  • Saito Y, Sharer LR, Epstein LG, Michaels J, Mintz M, Louder M, Golding K, Cvetkovich TA, Blumberg BM. Overexpression of nef as a marker for restricted HIV-1 infection of astrocytes in postmortem pediatric central nervous tissues. Neurology. 1994 Mar;44(3 Pt 1):474–481. [PubMed]
  • Salahuddin SZ, Markham PD, Wong-Staal F, Franchini G, Kalyanaraman VS, Gallo RC. Restricted expression of human T-cell leukemia--lymphoma virus (HTLV) in transformed human umbilical cord blood lymphocytes. Virology. 1983 Aug;129(1):51–64. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Schwartz S, Campbell M, Nasioulas G, Harrison J, Felber BK, Pavlakis GN. Mutational inactivation of an inhibitory sequence in human immunodeficiency virus type 1 results in Rev-independent gag expression. J Virol. 1992 Dec;66(12):7176–7182. [PMC free article] [PubMed]
  • Schwartz S, Felber BK, Benko DM, Fenyö EM, Pavlakis GN. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):2519–2529. [PMC free article] [PubMed]
  • Schwartz S, Felber BK, Pavlakis GN. Distinct RNA sequences in the gag region of human immunodeficiency virus type 1 decrease RNA stability and inhibit expression in the absence of Rev protein. J Virol. 1992 Jan;66(1):150–159. [PMC free article] [PubMed]
  • Schwartz S, Felber BK, Pavlakis GN. Mechanism of translation of monocistronic and multicistronic human immunodeficiency virus type 1 mRNAs. Mol Cell Biol. 1992 Jan;12(1):207–219. [PMC free article] [PubMed]
  • Shahabuddin M, Volsky B, Kim H, Sakai K, Volsky DJ. Regulated expression of human immunodeficiency virus type 1 in human glial cells: induction of dormant virus. Pathobiology. 1992;60(4):195–205. [PubMed]
  • Solomin L, Felber BK, Pavlakis GN. Different sites of interaction for Rev, Tev, and Rex proteins within the Rev-responsive element of human immunodeficiency virus type 1. J Virol. 1990 Dec;64(12):6010–6017. [PMC free article] [PubMed]
  • Tautz D, Renz M. An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem. 1983 Jul 1;132(1):14–19. [PubMed]
  • Tersmette M, Gruters RA, de Wolf F, de Goede RE, Lange JM, Schellekens PT, Goudsmit J, Huisman HG, Miedema F. Evidence for a role of virulent human immunodeficiency virus (HIV) variants in the pathogenesis of acquired immunodeficiency syndrome: studies on sequential HIV isolates. J Virol. 1989 May;63(5):2118–2125. [PMC free article] [PubMed]
  • Tornatore C, Chandra R, Berger JR, Major EO. HIV-1 infection of subcortical astrocytes in the pediatric central nervous system. Neurology. 1994 Mar;44(3 Pt 1):481–487. [PubMed]
  • Tornatore C, Meyers K, Atwood W, Conant K, Major E. Temporal patterns of human immunodeficiency virus type 1 transcripts in human fetal astrocytes. J Virol. 1994 Jan;68(1):93–102. [PMC free article] [PubMed]
  • Tornatore C, Nath A, Amemiya K, Major EO. Persistent human immunodeficiency virus type 1 infection in human fetal glial cells reactivated by T-cell factor(s) or by the cytokines tumor necrosis factor alpha and interleukin-1 beta. J Virol. 1991 Nov;65(11):6094–6100. [PMC free article] [PubMed]
  • Trono D, Baltimore D. A human cell factor is essential for HIV-1 Rev action. EMBO J. 1990 Dec;9(12):4155–4160. [PMC free article] [PubMed]
  • Vaishnav YN, Wong-Staal F. The biochemistry of AIDS. Annu Rev Biochem. 1991;60:577–630. [PubMed]
  • Werner T, Ferroni S, Saermark T, Brack-Werner R, Banati RB, Mager R, Steinaa L, Kreutzberg GW, Erfle V. HIV-1 Nef protein exhibits structural and functional similarity to scorpion peptides interacting with K+ channels. AIDS. 1991 Nov;5(11):1301–1308. [PubMed]
  • Westermark B, Pontén J, Hugosson R. Determinants for the establishment of permanent tissue culture lines from human gliomas. Acta Pathol Microbiol Scand A. 1973 Nov;81(6):791–805. [PubMed]
  • Winslow BJ, Pomerantz RJ, Bagasra O, Trono D. HIV-1 latency due to the site of proviral integration. Virology. 1993 Oct;196(2):849–854. [PubMed]
  • Winslow BJ, Trono D. The blocks to human immunodeficiency virus type 1 Tat and Rev functions in mouse cell lines are independent. J Virol. 1993 Apr;67(4):2349–2354. [PMC free article] [PubMed]
  • Zapp ML, Hope TJ, Parslow TG, Green MR. Oligomerization and RNA binding domains of the type 1 human immunodeficiency virus Rev protein: a dual function for an arginine-rich binding motif. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7734–7738. [PMC free article] [PubMed]

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