• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. Sep 1997; 71(9): 6641–6649.
PMCID: PMC191942

Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector.

Abstract

The identification of monogenic and complex genes responsible for neurological disorders requires new approaches for delivering therapeutic protein genes to significant numbers of cells in the central nervous system. A lentivirus-based vector capable of infecting dividing and quiescent cells was investigated in vivo by injecting highly concentrated viral vector stock into the striatum and hippocampus of adult rats. Control brains were injected with a Moloney murine leukemia virus, adenovirus, or adeno-associated virus vector. The volumes of the areas containing transduced cells and the transduced-cell densities were stereologically determined to provide a basis for comparison among different viral vectors and variants of the viral vector stocks. The efficiency of infection by the lentivirus vector was improved by deoxynucleoside triphosphate pretreatment of the vector and was reduced following mutation of integrase and the Vpr-matrix protein complex involved in the nuclear translocation of the preintegration complex. The lentivirus vector system was able to efficiently and stably infect quiescent cells in the primary injection site with transgene expression for over 6 months. Triple labeling showed that 88.7% of striatal cells transduced by the lentivirus vector were terminally differentiated neurons.

Full Text

The Full Text of this article is available as a PDF (558K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Afione SA, Conrad CK, Kearns WG, Chunduru S, Adams R, Reynolds TC, Guggino WB, Cutting GR, Carter BJ, Flotte TR. In vivo model of adeno-associated virus vector persistence and rescue. J Virol. 1996 May;70(5):3235–3241. [PMC free article] [PubMed]
  • Akli S, Caillaud C, Vigne E, Stratford-Perricaudet LD, Poenaru L, Perricaudet M, Kahn A, Peschanski MR. Transfer of a foreign gene into the brain using adenovirus vectors. Nat Genet. 1993 Mar;3(3):224–228. [PubMed]
  • Bajocchi G, Feldman SH, Crystal RG, Mastrangeli A. Direct in vivo gene transfer to ependymal cells in the central nervous system using recombinant adenovirus vectors. Nat Genet. 1993 Mar;3(3):229–234. [PubMed]
  • Baskar JF, Smith PP, Nilaver G, Jupp RA, Hoffmann S, Peffer NJ, Tenney DJ, Colberg-Poley AM, Ghazal P, Nelson JA. The enhancer domain of the human cytomegalovirus major immediate-early promoter determines cell type-specific expression in transgenic mice. J Virol. 1996 May;70(5):3207–3214. [PMC free article] [PubMed]
  • Berns KI, Linden RM. The cryptic life style of adeno-associated virus. Bioessays. 1995 Mar;17(3):237–245. [PubMed]
  • Brubaker JO, Thompson CM, Morrison LA, Knipe DM, Siber GR, Finberg RW. Th1-associated immune responses to beta-galactosidase expressed by a replication-defective herpes simplex virus. J Immunol. 1996 Aug 15;157(4):1598–1604. [PubMed]
  • Burns JC, Friedmann T, Driever W, Burrascano M, Yee JK. Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8033–8037. [PMC free article] [PubMed]
  • Byrnes AP, MacLaren RE, Charlton HM. Immunological instability of persistent adenovirus vectors in the brain: peripheral exposure to vector leads to renewed inflammation, reduced gene expression, and demyelination. J Neurosci. 1996 May 1;16(9):3045–3055. [PubMed]
  • Byrnes AP, Rusby JE, Wood MJ, Charlton HM. Adenovirus gene transfer causes inflammation in the brain. Neuroscience. 1995 Jun;66(4):1015–1024. [PubMed]
  • Byrnes AP, Wood MJ, Charlton HM. Role of T cells in inflammation caused by adenovirus vectors in the brain. Gene Ther. 1996 Jul;3(7):644–651. [PubMed]
  • Charneau P, Mirambeau G, Roux P, Paulous S, Buc H, Clavel F. HIV-1 reverse transcription. A termination step at the center of the genome. J Mol Biol. 1994 Sep 2;241(5):651–662. [PubMed]
  • Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PMC free article] [PubMed]
  • Chiocca EA, Choi BB, Cai WZ, DeLuca NA, Schaffer PA, DiFiglia M, Breakefield XO, Martuza RL. Transfer and expression of the lacZ gene in rat brain neurons mediated by herpes simplex virus mutants. New Biol. 1990 Aug;2(8):739–746. [PubMed]
  • Davidson BL, Doran SE, Shewach DS, Latta JM, Hartman JW, Roessler BJ. Expression of Escherichia coli beta-galactosidase and rat HPRT in the CNS of Macaca mulatta following adenoviral mediated gene transfer. Exp Neurol. 1994 Feb;125(2):258–267. [PubMed]
  • Engelhardt JF, Ye X, Doranz B, Wilson JM. Ablation of E2A in recombinant adenoviruses improves transgene persistence and decreases inflammatory response in mouse liver. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6196–6200. [PMC free article] [PubMed]
  • Esiri MM. Herpes simplex encephalitis. An immunohistological study of the distribution of viral antigen within the brain. J Neurol Sci. 1982 May;54(2):209–226. [PubMed]
  • Ferrari FK, Samulski T, Shenk T, Samulski RJ. Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors. J Virol. 1996 May;70(5):3227–3234. [PMC free article] [PubMed]
  • Friedman B, Hockfield S, Black JA, Woodruff KA, Waxman SG. In situ demonstration of mature oligodendrocytes and their processes: an immunocytochemical study with a new monoclonal antibody, rip. Glia. 1989;2(5):380–390. [PubMed]
  • Friedmann T. Gene therapy for disorders of the CNS. Gene Ther. 1994;1 (Suppl 1):S47–S48. [PubMed]
  • Friedmann T. The promise and overpromise of human gene therapy. Gene Ther. 1994 Jul;1(4):217–218. [PubMed]
  • Gallay P, Stitt V, Mundy C, Oettinger M, Trono D. Role of the karyopherin pathway in human immunodeficiency virus type 1 nuclear import. J Virol. 1996 Feb;70(2):1027–1032. [PMC free article] [PubMed]
  • Gallay P, Swingler S, Aiken C, Trono D. HIV-1 infection of nondividing cells: C-terminal tyrosine phosphorylation of the viral matrix protein is a key regulator. Cell. 1995 Feb 10;80(3):379–388. [PubMed]
  • Gallay P, Swingler S, Song J, Bushman F, Trono D. HIV nuclear import is governed by the phosphotyrosine-mediated binding of matrix to the core domain of integrase. Cell. 1995 Nov 17;83(4):569–576. [PubMed]
  • Gao WY, Cara A, Gallo RC, Lori F. Low levels of deoxynucleotides in peripheral blood lymphocytes: a strategy to inhibit human immunodeficiency virus type 1 replication. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8925–8928. [PMC free article] [PubMed]
  • Goodman S, Xiao X, Donahue RE, Moulton A, Miller J, Walsh C, Young NS, Samulski RJ, Nienhuis AW. Recombinant adeno-associated virus-mediated gene transfer into hematopoietic progenitor cells. Blood. 1994 Sep 1;84(5):1492–1500. [PubMed]
  • Halbert CL, Alexander IE, Wolgamot GM, Miller AD. Adeno-associated virus vectors transduce primary cells much less efficiently than immortalized cells. J Virol. 1995 Mar;69(3):1473–1479. [PMC free article] [PubMed]
  • Huang Q, Vonsattel JP, Schaffer PA, Martuza RL, Breakefield XO, DiFiglia M. Introduction of a foreign gene (Escherichia coli lacZ) into rat neostriatal neurons using herpes simplex virus mutants: a light and electron microscopic study. Exp Neurol. 1992 Mar;115(3):303–316. [PubMed]
  • Jolly D. Viral vector systems for gene therapy. Cancer Gene Ther. 1994 Mar;1(1):51–64. [PubMed]
  • Kaplitt MG, Leone P, Samulski RJ, Xiao X, Pfaff DW, O'Malley KL, During MJ. Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain. Nat Genet. 1994 Oct;8(2):148–154. [PubMed]
  • Kass-Eisler A, Falck-Pedersen E, Elfenbein DH, Alvira M, Buttrick PM, Leinwand LA. The impact of developmental stage, route of administration and the immune system on adenovirus-mediated gene transfer. Gene Ther. 1994 Nov;1(6):395–402. [PubMed]
  • Knowles MR, Hohneker KW, Zhou Z, Olsen JC, Noah TL, Hu PC, Leigh MW, Engelhardt JF, Edwards LJ, Jones KR, et al. A controlled study of adenoviral-vector-mediated gene transfer in the nasal epithelium of patients with cystic fibrosis. N Engl J Med. 1995 Sep 28;333(13):823–831. [PubMed]
  • Kotin RM, Berns KI. Organization of adeno-associated virus DNA in latently infected Detroit 6 cells. Virology. 1989 Jun;170(2):460–467. [PubMed]
  • Kuo H, Ingram DK, Crystal RG, Mastrangeli A. Retrograde transfer of replication deficient recombinant adenovirus vector in the central nervous system for tracing studies. Brain Res. 1995 Dec 24;705(1-2):31–38. [PubMed]
  • Leavitt AD, Robles G, Alesandro N, Varmus HE. Human immunodeficiency virus type 1 integrase mutants retain in vitro integrase activity yet fail to integrate viral DNA efficiently during infection. J Virol. 1996 Feb;70(2):721–728. [PMC free article] [PubMed]
  • Lebkowski JS, McNally MM, Okarma TB, Lerch LB. Adeno-associated virus: a vector system for efficient introduction and integration of DNA into a variety of mammalian cell types. Mol Cell Biol. 1988 Oct;8(10):3988–3996. [PMC free article] [PubMed]
  • Leeds JM, Mathews CK. Cell cycle-dependent effects on deoxyribonucleotide and DNA labeling by nucleoside precursors in mammalian cells. Mol Cell Biol. 1987 Jan;7(1):532–534. [PMC free article] [PubMed]
  • Leeds JM, Slabaugh MB, Mathews CK. DNA precursor pools and ribonucleotide reductase activity: distribution between the nucleus and cytoplasm of mammalian cells. Mol Cell Biol. 1985 Dec;5(12):3443–3450. [PMC free article] [PubMed]
  • Lewis P, Hensel M, Emerman M. Human immunodeficiency virus infection of cells arrested in the cell cycle. EMBO J. 1992 Aug;11(8):3053–3058. [PMC free article] [PubMed]
  • Meyerhans A, Vartanian JP, Hultgren C, Plikat U, Karlsson A, Wang L, Eriksson S, Wain-Hobson S. Restriction and enhancement of human immunodeficiency virus type 1 replication by modulation of intracellular deoxynucleoside triphosphate pools. J Virol. 1994 Jan;68(1):535–540. [PMC free article] [PubMed]
  • Miller AD. Cell-surface receptors for retroviruses and implications for gene transfer. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11407–11413. [PMC free article] [PubMed]
  • Miller AD, Law MF, Verma IM. Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene. Mol Cell Biol. 1985 Mar;5(3):431–437. [PMC free article] [PubMed]
  • Miller DG, Adam MA, Miller AD. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol. 1990 Aug;10(8):4239–4242. [PMC free article] [PubMed]
  • Miller JL, Donahue RE, Sellers SE, Samulski RJ, Young NS, Nienhuis AW. Recombinant adeno-associated virus (rAAV)-mediated expression of a human gamma-globin gene in human progenitor-derived erythroid cells. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10183–10187. [PMC free article] [PubMed]
  • Mullen RJ, Buck CR, Smith AM. NeuN, a neuronal specific nuclear protein in vertebrates. Development. 1992 Sep;116(1):201–211. [PubMed]
  • Mulligan RC. The basic science of gene therapy. Science. 1993 May 14;260(5110):926–932. [PubMed]
  • Muzyczka N. Use of adeno-associated virus as a general transduction vector for mammalian cells. Curr Top Microbiol Immunol. 1992;158:97–129. [PubMed]
  • Naldini L, Blömer U, Gage FH, Trono D, Verma IM. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11382–11388. [PMC free article] [PubMed]
  • Naldini L, Blömer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science. 1996 Apr 12;272(5259):263–267. [PubMed]
  • Naviaux RK, Costanzi E, Haas M, Verma IM. The pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses. J Virol. 1996 Aug;70(8):5701–5705. [PMC free article] [PubMed]
  • Palmer TD, Rosman GJ, Osborne WR, Miller AD. Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1330–1334. [PMC free article] [PubMed]
  • Pear WS, Nolan GP, Scott ML, Baltimore D. Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8392–8396. [PMC free article] [PubMed]
  • Peterson DA, Lucidi-Phillipi CA, Eagle KL, Gage FH. Perforant path damage results in progressive neuronal death and somal atrophy in layer II of entorhinal cortex and functional impairment with increasing postdamage age. J Neurosci. 1994 Nov;14(11 Pt 2):6872–6885. [PubMed]
  • Porter CD, Collins MK, Tailor CS, Parkar MH, Cosset FL, Weiss RA, Takeuchi Y. Comparison of efficiency of infection of human gene therapy target cells via four different retroviral receptors. Hum Gene Ther. 1996 May 20;7(8):913–919. [PubMed]
  • Reddy GP, Fager RS. Replitase: a complex integrating dNTP synthesis and DNA replication. Crit Rev Eukaryot Gene Expr. 1993;3(4):255–277. [PubMed]
  • Richardson WD, Westphal H. Requirement for either early region 1a or early region 1b adenovirus gene products in the helper effect for adeno-associated virus. J Virol. 1984 Aug;51(2):404–410. [PMC free article] [PubMed]
  • Samulski RJ, Chang LS, Shenk T. Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression. J Virol. 1989 Sep;63(9):3822–3828. [PMC free article] [PubMed]
  • Scharfmann R, Axelrod JH, Verma IM. Long-term in vivo expression of retrovirus-mediated gene transfer in mouse fibroblast implants. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4626–4630. [PMC free article] [PubMed]
  • Sterio DC. The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc. 1984 May;134(Pt 2):127–136. [PubMed]
  • St Louis D, Verma IM. An alternative approach to somatic cell gene therapy. Proc Natl Acad Sci U S A. 1988 May;85(9):3150–3154. [PMC free article] [PubMed]
  • Trapnell BC, Gorziglia M. Gene therapy using adenoviral vectors. Curr Opin Biotechnol. 1994 Dec;5(6):617–625. [PubMed]
  • Verma IM. Gene therapy: hopes, hypes, and hurdles. Mol Med. 1994 Nov;1(1):2–3. [PMC free article] [PubMed]
  • von Schwedler U, Kornbluth RS, Trono D. The nuclear localization signal of the matrix protein of human immunodeficiency virus type 1 allows the establishment of infection in macrophages and quiescent T lymphocytes. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6992–6996. [PMC free article] [PubMed]
  • West MH, Trempe JP, Tratschin JD, Carter BJ. Gene expression in adeno-associated virus vectors: the effects of chimeric mRNA structure, helper virus, and adenovirus VA1 RNA. Virology. 1987 Sep;160(1):38–47. [PubMed]
  • Wood MJ, Byrnes AP, Kaplitt MG, Pfaff DW, Rabkin SD, Charlton HM. Specific patterns of defective HSV-1 gene transfer in the adult central nervous system: implications for gene targeting. Exp Neurol. 1994 Nov;130(1):127–140. [PubMed]
  • Yang Y, Nunes FA, Berencsi K, Gönczöl E, Engelhardt JF, Wilson JM. Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosis. Nat Genet. 1994 Jul;7(3):362–369. [PubMed]
  • Závada J. Pseudotypes of vesicular stomatitis virus with the coat of murine leukaemia and of avian myeloblastosis viruses. J Gen Virol. 1972 Jun;15(3):183–191. [PubMed]
  • Zhang H, Duan LX, Dornadula G, Pomerantz RJ. Increasing transduction efficiency of recombinant murine retrovirus vectors by initiation of endogenous reverse transcription: potential utility for genetic therapies. J Virol. 1995 Jun;69(6):3929–3932. [PMC free article] [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...