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. 1994 May;68(5):3401–3409. doi: 10.1128/jvi.68.5.3401-3409.1994

Microglial infection by a neurovirulent murine retrovirus results in defective processing of envelope protein and intracellular budding of virus particles.

W P Lynch 1, W J Brown 1, G J Spangrude 1, J L Portis 1
PMCID: PMC236834  PMID: 8151801

Abstract

The observation of murine retrovirus infection of microglial cells in brain regions expressing spongiform neurodegenerative changes suggests that these cells may play an important role in pathogenesis. To evaluate this potential in vitro, murine microglial cells were infected in mixed glial cultures with the highly neurovirulent murine retrovirus, FrCasE. The microglia were then isolated from the mixed cultures on the basis of their differential adherence and shown to be approximately 98% pure. The infected microglia expressed viral envelope protein at the plasma membrane, while viral budding was primarily intracellular. Evaluation of the viral envelope protein by immunoblotting indicated that the immunoreactive species produced was exclusively a 90-kDa precursor protein. Very little of the envelope protein was associated with particles released from these cells, and viral titers in the culture supernatant were low. Interestingly, these cells were still capable of infecting permissive target cells when seeded as infectious centers. This partially defective infection of microglial cells suggests a potential cellular means by which a neurovirulent retrovirus could disrupt normal microglia and in turn central nervous system motor system functioning.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  2. Baszler T. V., Zachary J. F. Murine retroviral neurovirulence correlates with an enhanced ability ofvirus to infect selectively, replicate in, and activate resident microglial cells. Am J Pathol. 1991 Mar;138(3):655–671. [PMC free article] [PubMed] [Google Scholar]
  3. Baszler T. V., Zachary J. F. Murine retroviral-induced spongiform neuronal degeneration parallels resident microglial cell infection: ultrastructural findings. Lab Invest. 1990 Nov;63(5):612–623. [PubMed] [Google Scholar]
  4. Bignami A., Eng L. F., Dahl D., Uyeda C. T. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 1972 Aug 25;43(2):429–435. doi: 10.1016/0006-8993(72)90398-8. [DOI] [PubMed] [Google Scholar]
  5. Brooks B. R., Swarz J. R., Narayan O., Johnson R. T. Murine neurotropic retrovirus spongiform polioencephalomyelopathy: acceleration of disease by virus inoculum concentration. Infect Immun. 1979 Feb;23(2):540–544. doi: 10.1128/iai.23.2.540-544.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown W. J., Farquhar M. G. Immunoperoxidase methods for the localization of antigens in cultured cells and tissue sections by electron microscopy. Methods Cell Biol. 1989;31:553–569. doi: 10.1016/s0091-679x(08)61626-x. [DOI] [PubMed] [Google Scholar]
  7. Chesebro B., Britt W., Evans L., Wehrly K., Nishio J., Cloyd M. Characterization of monoclonal antibodies reactive with murine leukemia viruses: use in analysis of strains of friend MCF and Friend ecotropic murine leukemia virus. Virology. 1983 May;127(1):134–148. doi: 10.1016/0042-6822(83)90378-1. [DOI] [PubMed] [Google Scholar]
  8. Chesebro B., Wehrly K., Watson K., Chesebro K. Murine leukemia virus infectious centers are dependent on the rate of virus production by infected cells. Virology. 1978 Jan;84(1):222–226. doi: 10.1016/0042-6822(78)90239-8. [DOI] [PubMed] [Google Scholar]
  9. Czub M., Czub S., McAtee F. J., Portis J. L. Age-dependent resistance to murine retrovirus-induced spongiform neurodegeneration results from central nervous system-specific restriction of virus replication. J Virol. 1991 May;65(5):2539–2544. doi: 10.1128/jvi.65.5.2539-2544.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Czub M., McAtee F. J., Portis J. L. Murine retrovirus-induced spongiform encephalomyelopathy: host and viral factors which determine the length of the incubation period. J Virol. 1992 Jun;66(6):3298–3305. doi: 10.1128/jvi.66.6.3298-3305.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dong J. Y., Dubay J. W., Perez L. G., Hunter E. Mutations within the proteolytic cleavage site of the Rous sarcoma virus glycoprotein define a requirement for dibasic residues for intracellular cleavage. J Virol. 1992 Feb;66(2):865–874. doi: 10.1128/jvi.66.2.865-874.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gardner M. B., Henderson B. E., Officer J. E., Rongey R. W., Parker J. C., Oliver C., Estes J. D., Huebner R. J. A spontaneous lower motor neuron disease apparently caused by indigenous type-C RNA virus in wild mice. J Natl Cancer Inst. 1973 Oct;51(4):1243–1254. doi: 10.1093/jnci/51.4.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gendelman H. E., Orenstein J. M., Martin M. A., Ferrua C., Mitra R., Phipps T., Wahl L. A., Lane H. C., Fauci A. S., Burke D. S. Efficient isolation and propagation of human immunodeficiency virus on recombinant colony-stimulating factor 1-treated monocytes. J Exp Med. 1988 Apr 1;167(4):1428–1441. doi: 10.1084/jem.167.4.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Giulian D., Baker T. J. Characterization of ameboid microglia isolated from developing mammalian brain. J Neurosci. 1986 Aug;6(8):2163–2178. doi: 10.1523/JNEUROSCI.06-08-02163.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gliniak B. C., Kozak S. L., Jones R. T., Kabat D. Disulfide bonding controls the processing of retroviral envelope glycoproteins. J Biol Chem. 1991 Dec 5;266(34):22991–22997. [PubMed] [Google Scholar]
  16. Gravel C., Kay D. G., Jolicoeur P. Identification of the infected target cell type in spongiform myeloencephalopathy induced by the neurotropic Cas-Br-E murine leukemia virus. J Virol. 1993 Nov;67(11):6648–6658. doi: 10.1128/jvi.67.11.6648-6658.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jolicoeur P., Nicolaiew N., DesGroseillers L., Rassart E. Molecular cloning of infectious viral DNA from ecotropic neurotropic wild mouse retrovirus. J Virol. 1983 Mar;45(3):1159–1163. doi: 10.1128/jvi.45.3.1159-1163.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kay D. G., Gravel C., Pothier F., Laperrière A., Robitaille Y., Jolicoeur P. Neurological disease induced in transgenic mice expressing the env gene of the Cas-Br-E murine retrovirus. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4538–4542. doi: 10.1073/pnas.90.10.4538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kayman S. C., Kopelman R., Projan S., Kinney D. M., Pinter A. Mutational analysis of N-linked glycosylation sites of Friend murine leukemia virus envelope protein. J Virol. 1991 Oct;65(10):5323–5332. doi: 10.1128/jvi.65.10.5323-5332.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kim J. W., Cunningham J. M. N-linked glycosylation of the receptor for murine ecotropic retroviruses is altered in virus-infected cells. J Biol Chem. 1993 Aug 5;268(22):16316–16320. [PubMed] [Google Scholar]
  21. Kozak S. L., Kabat D. Ping-pong amplification of a retroviral vector achieves high-level gene expression: human growth hormone production. J Virol. 1990 Jul;64(7):3500–3508. doi: 10.1128/jvi.64.7.3500-3508.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Linial M., Fenno J., Burnette W. N., Rohrschneider L. Synthesis and processing of viral glycoproteins in two nonconditional mutants of Rous sarcoma virus. J Virol. 1980 Oct;36(1):280–290. doi: 10.1128/jvi.36.1.280-290.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lynch W. P., Czub S., McAtee F. J., Hayes S. F., Portis J. L. Murine retrovirus-induced spongiform encephalopathy: productive infection of microglia and cerebellar neurons in accelerated CNS disease. Neuron. 1991 Sep;7(3):365–379. doi: 10.1016/0896-6273(91)90289-c. [DOI] [PubMed] [Google Scholar]
  24. Lynch W. P., Portis J. L. Murine retrovirus-induced spongiform encephalopathy: disease expression is dependent on postnatal development of the central nervous system. J Virol. 1993 May;67(5):2601–2610. doi: 10.1128/jvi.67.5.2601-2610.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McAtee F. J., Portis J. L. Monoclonal antibodies specific for wild mouse neurotropic retrovirus: detection of comparable levels of virus replication in mouse strains susceptible and resistant to paralytic disease. J Virol. 1985 Dec;56(3):1018–1022. doi: 10.1128/jvi.56.3.1018-1022.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. McComb R. D., Jones T. R., Pizzo S. V., Bigner D. D. Specificity and sensitivity of immunohistochemical detection of factor VIII/von Willebrand factor antigen in formalin-fixed paraffin-embedded tissue. J Histochem Cytochem. 1982 Apr;30(4):371–377. doi: 10.1177/30.4.6801111. [DOI] [PubMed] [Google Scholar]
  27. Michaels J., Price R. W., Rosenblum M. K. Microglia in the giant cell encephalitis of acquired immune deficiency syndrome: proliferation, infection and fusion. Acta Neuropathol. 1988;76(4):373–379. doi: 10.1007/BF00686974. [DOI] [PubMed] [Google Scholar]
  28. Miller A. D., Law M. F., Verma I. M. 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. doi: 10.1128/mcb.5.3.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Orenstein J. M., Meltzer M. S., Phipps T., Gendelman H. E. Cytoplasmic assembly and accumulation of human immunodeficiency virus types 1 and 2 in recombinant human colony-stimulating factor-1-treated human monocytes: an ultrastructural study. J Virol. 1988 Aug;62(8):2578–2586. doi: 10.1128/jvi.62.8.2578-2586.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Portis J. L., Czub S., Garon C. F., McAtee F. J. Neurodegenerative disease induced by the wild mouse ecotropic retrovirus is markedly accelerated by long terminal repeat and gag-pol sequences from nondefective Friend murine leukemia virus. J Virol. 1990 Apr;64(4):1648–1656. doi: 10.1128/jvi.64.4.1648-1656.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Portis J. L. Wild mouse retrovirus: pathogenesis. Curr Top Microbiol Immunol. 1990;160:11–27. doi: 10.1007/978-3-642-75267-4_2. [DOI] [PubMed] [Google Scholar]
  32. Poss M. L., Dow S. W., Hoover E. A. Cell-specific envelope glycosylation distinguishes FIV glycoproteins produced in cytopathically and noncytopathically infected cells. Virology. 1992 May;188(1):25–32. doi: 10.1016/0042-6822(92)90731-4. [DOI] [PubMed] [Google Scholar]
  33. Poss M. L., Quackenbush S. L., Mullins J. I., Hoover E. A. Characterization and significance of delayed processing of the feline leukemia virus FeLV-FAIDS envelope glycoprotein. J Virol. 1990 Sep;64(9):4338–4345. doi: 10.1128/jvi.64.9.4338-4345.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Raff M. C., Mirsky R., Fields K. L., Lisak R. P., Dorfman S. H., Silberberg D. H., Gregson N. A., Leibowitz S., Kennedy M. C. Galactocerebroside is a specific cell-surface antigenic marker for oligodendrocytes in culture. Nature. 1978 Aug 24;274(5673):813–816. [PubMed] [Google Scholar]
  35. Rassart E., Nelbach L., Jolicoeur P. Cas-Br-E murine leukemia virus: sequencing of the paralytogenic region of its genome and derivation of specific probes to study its origin and the structure of its recombinant genomes in leukemic tissues. J Virol. 1986 Dec;60(3):910–919. doi: 10.1128/jvi.60.3.910-919.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Richardson A., Hao C., Fedoroff S. Microglia progenitor cells: a subpopulation in cultures of mouse neopallial astroglia. Glia. 1993 Jan;7(1):25–33. doi: 10.1002/glia.440070107. [DOI] [PubMed] [Google Scholar]
  37. Sharpe A. H., Jaenisch R., Ruprecht R. M. Retroviruses and mouse embryos: a rapid model for neurovirulence and transplacental antiviral therapy. Science. 1987 Jun 26;236(4809):1671–1674. doi: 10.1126/science.3037694. [DOI] [PubMed] [Google Scholar]
  38. Shikova E., Lin Y. C., Saha K., Brooks B. R., Wong P. K. Correlation of specific virus-astrocyte interactions and cytopathic effects induced by ts1, a neurovirulent mutant of Moloney murine leukemia virus. J Virol. 1993 Mar;67(3):1137–1147. doi: 10.1128/jvi.67.3.1137-1147.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Springer T., Galfrè G., Secher D. S., Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978 Aug;8(8):539–551. doi: 10.1002/eji.1830080802. [DOI] [PubMed] [Google Scholar]
  40. Springer T., Galfré G., Secher D. S., Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979 Apr;9(4):301–306. doi: 10.1002/eji.1830090410. [DOI] [PubMed] [Google Scholar]
  41. Streit W. J., Kreutzberg G. W. Lectin binding by resting and reactive microglia. J Neurocytol. 1987 Apr;16(2):249–260. doi: 10.1007/BF01795308. [DOI] [PubMed] [Google Scholar]
  42. Vazeux R., Brousse N., Jarry A., Henin D., Marche C., Vedrenne C., Mikol J., Wolff M., Michon C., Rozenbaum W. AIDS subacute encephalitis. Identification of HIV-infected cells. Am J Pathol. 1987 Mar;126(3):403–410. [PMC free article] [PubMed] [Google Scholar]
  43. Watkins B. A., Dorn H. H., Kelly W. B., Armstrong R. C., Potts B. J., Michaels F., Kufta C. V., Dubois-Dalcq M. Specific tropism of HIV-1 for microglial cells in primary human brain cultures. Science. 1990 Aug 3;249(4968):549–553. doi: 10.1126/science.2200125. [DOI] [PubMed] [Google Scholar]
  44. Wiley C. A., Schrier R. D., Nelson J. A., Lampert P. W., Oldstone M. B. Cellular localization of human immunodeficiency virus infection within the brains of acquired immune deficiency syndrome patients. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7089–7093. doi: 10.1073/pnas.83.18.7089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wong P. K., Soong M. M., MacLeod R., Gallick G. E., Yuen P. H. A group of temperature-sensitive mutants of Moloney leukemia virus which is defective in cleavage of env precursor polypeptide in infected cells also induces hind-limb paralysis in newborn CFW/D mice. Virology. 1983 Mar;125(2):513–518. doi: 10.1016/0042-6822(83)90225-8. [DOI] [PubMed] [Google Scholar]
  46. Yuen P. H., Malehorn D., Knupp C., Wong P. K. A 1.6-kilobase-pair fragment in the genome of the ts1 mutant of Moloney murine leukemia virus TB that is associated with temperature sensitivity, nonprocessing of Pr80env, and paralytogenesis. J Virol. 1985 May;54(2):364–373. doi: 10.1128/jvi.54.2.364-373.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yuen P. H., Tzeng E., Knupp C., Wong P. K. The neurovirulent determinants of ts1, a paralytogenic mutant of Moloney murine leukemia virus TB, are localized in at least two functionally distinct regions of the genome. J Virol. 1986 Jul;59(1):59–65. doi: 10.1128/jvi.59.1.59-65.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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