Archives of Virology

, Volume 111, Issue 1–2, pp 87–101 | Cite as

The budding of defective human immunodeficiency virus type 1 (HIV-1) particles from cell clones persistently infected with HIV-1

  • T. Goto
  • K. Ikuta
  • J. J. Zhang
  • C. Morita
  • K. Sano
  • M. Komatsu
  • H. Fujita
  • S. Kato
  • M. Nakai
Original Papers


Three cell clones producing large numbers of infectious or noninfectious particles of human immunodeficiency virus type 1 (HIV-1), designated M 10/LAV-2, M 16/LAV-3, and MT/LAV-17, were isolated from persistently HIV-1-infected MT-4 cells. In M 10/LAV-2, the HIV-1 proteins were defective in the cleavage ofgag precursor protein, and the particles were doughnutshaped with a double-ring structure. These particles were produced by budding at the cell surface from crescentic structures followed by the formation of double-ring structures. The viral proteins in M 16/LAV-3 were defective in the cleavage ofenv precursor protein. The morphology of the virus particles was intact, and an electron dense bar-shaped core was seen inside a single-ring enveloped structure. The intact particles were released from the cell surface by a budding process in which crescent shape structures first appeared at the cell membrane, then subsequently just before release matured to a complete structure with an electron dense core. In MT/LAV-17, the synthesis of HIV-1 proteins was normal, and the particles were teardrop-shaped with an intact core structure. These particles were produced by budding with an electron dense core at the cell surface.

Thus, it was suggested that the morphological maturation of HIV-1 particles was completed just before release from the cell surface in several cell clones producing HIV-1 particles of different morphology.


Precursor Protein Cell Clone Shape Structure Intact Core Crescent Shape 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Barré-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, Dauguet C, Axler-Blin C, Vézinet-Brun F, Rouzioux C, Rozenbaum W, Montagnier L (1983) Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 220: 868–871Google Scholar
  2. 2.
    Dalgleish AG, Beverley PCL, Clapham PR, Crawford DH, Greaves MF, Weiss RA (1984) The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus. Nature 312: 763–767Google Scholar
  3. 3.
    Fitting T, Kabat D (1982) Evidence for a glycoprotein “signal” involved in transport between subcellular organelles. Two membrane glycoproteins encoded by murine leukemia virus reach the cell surface at different rates. J Biol Chem 257: 14011–14017Google Scholar
  4. 4.
    Gazdar AF, Phillips LA, Sarma PS, Peebles PT, Chopra HC (1971) Presence of sarcoma genome in a “non-infectious” mammalian virus. Nature [New Biol] 234: 69–72Google Scholar
  5. 5.
    Gelderblom HR, Hausmann EHS, Özel M, Pauli G, Koch MA (1987) Fine structure of human immunodeficiency virus (HIV) and immunolocalization of structural proteins. Virology 156: 171–176Google Scholar
  6. 6.
    Goto T, Harada S, Yamamoto N, Nakai M (1988) Entry of human immunodeficiency virus (HIV) into MT-2, human T cell leukemia virus carrier cell line. Arch Virol 102: 29–38Google Scholar
  7. 7.
    Gumbiner B, Kelly RB (1982) Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell 28: 51–59Google Scholar
  8. 8.
    Harada S, Koyanagi Y, Yamamoto N (1985) Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. Science 229: 563–566Google Scholar
  9. 9.
    Henderson LE, Sowder R, Copeland TD, Smythers G, Oroszlan S (1984) Quantitative separation of murine leukemia virus proteins by reversed-phase high-pressure liquid chromatography reveals newly describedgag andenv cleavage products. J Virol 52: 492–500Google Scholar
  10. 10.
    Hoxie JA, Alpers JD, Rackowski JL, Huebner K, Haggarty BS, Cedarbaum AJ, Read JC (1986) Alterations in T4(CD4) protein and mRNA synthesis in cells infected with HIV. Science 234: 1123–1127Google Scholar
  11. 11.
    Ikuta K, Coward J, Luftig RB (1986) The effect cerulenin on the synthesis of the precursorgag polyprotein in defective murine leukemia and sarcoma virus producing cell lines. Virology 154: 207–213Google Scholar
  12. 12.
    Ikuta K, Imai H, Ueda S, Suehiro S, Yamamoto N, Kato S (1987) Amplification of human immunodeficiency virus production from a virus-producing cell line in culture at high cell density. Jpn J Cancer Res (Gann) 78: 643–647Google Scholar
  13. 13.
    Ikuta K, Morita C, Miyake S, Ito T, Okabayashi M, Sano K, Nakai M, Hirai K, Kato S (1989) Expression of human immunodeficiency virus type 1 (HIV-1)gag antigens on the surface of a cell line persistently infected with HIV-1 that highly expressed HIV-1 antigens. Virology 170: 408–417Google Scholar
  14. 14.
    Ikuta K, Morita C, Nakai M, Yamamoto N, Kato S (1988) Defective human immunodeficiency virus (HIV) particles produced by cloned cells of HTLV-I-carrying MT-4 cells persistently infected with HIV. Jpn J Cancer Res (Gann) 79: 418–423Google Scholar
  15. 15.
    Katoh I, Yoshinaka Y, Rein A, Shibuya M, Odaka T, Oroszlan S (1985) Murine leukemia virus maturation: protease region required for conversion from “immature” to “mature” core form and for virus infectivity. Virology 145: 280–292Google Scholar
  16. 16.
    Katsumoto T, Hattori N, Kurimura T (1987) Maturation of human immunodeficiency virus, strain LAV, in vitro. Intervirology 27: 148–153Google Scholar
  17. 17.
    Katsumoto T, Hattori N, Yamada O, Kurimura T (1988) Intermediate virions in maturation of human immunodeficiency virus, strain LAV. J Electron Microsc 37: 205–207Google Scholar
  18. 18.
    Klatzmann D, Barré-Sinoussi F, Nugeyre MT, Dauguet C, Vilmer E, Griscelli C, Brun-Vezinet F, Rouzioux C, Gluckman JC, Chermann JC, Montagnier L (1984) Selective tropism of lymphadenopathy associated virus (LAV) for helper-inducer T lymphocytes. Science 225: 59–63Google Scholar
  19. 19.
    Klatzmann D, Champagne E, Chamaret S, Gruest J, Guetard D, Hercend T, Gluckman JC, Montagnier L (1984) T-lymphocyte T4 molecule behaves as the receptor for human retrovirus LAV. Nature 312: 767–768Google Scholar
  20. 20.
    Koyanagi Y, Harada S, Yamamoto N (1985) Correlation between high susceptibility to AIDS virus and surface expression of OKT-4 antigen in HTLV-I-positive cell lines. Jpn J Cancer Res (Gann) 76: 799–802Google Scholar
  21. 21.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685Google Scholar
  22. 22.
    Lee NH, Sano K, Morales FE, Imagawa DT (1987) Sensitive reverse transcriptase assay to detect and quantitate human immunodeficiency virus. J Clin Microbiol 25: 1717–1721Google Scholar
  23. 23.
    Lifson JD, Reyes GR, McGrath MS, Stein BS, Engleman EG (1986) AIDS retrovirus induced cytopathology: giant cell formation and involvement of CD 4 antigen. Science 232: 1123–1127Google Scholar
  24. 24.
    Lu AH, Soong MM, Wong PKY (1979) Maturation of Moloney murine leukemia virus. Virology 93: 269–274Google Scholar
  25. 25.
    Luftig RB, Yoshinaka Y (1978) Rauscher leukemia virus populations enriched for “immature” virions contain increased amounts of P70, thegag gene product. J Virol 25: 416–421Google Scholar
  26. 26.
    Maddon PJ, Dalgleish AG, McDougal JS, Clapham PR, Weiss RA, Axel R (1986) The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell 47: 333–348Google Scholar
  27. 27.
    McDougal JS, Kennedy MS, Sligh JM, Cort SP, Mawle A, Nicholson JKA (1986) Binding of HTLV-III/LAV to T4+ T cells by a complex of the 110K viral protein and the T4 molecule. Science 231: 382–385Google Scholar
  28. 28.
    McDougal JS, Nicholson JKA, Cross GD, Cort SP, Kennedy MS, Mawle AC (1986) Binding of the human retrovirus HTLV-III/LAV/ARV/HIV to the CD4 (T4) molecule: conformation dependence, epitope mapping, antibody inhibition, and potential for idiotypic mimicry. J Immunol 137: 2937–2944Google Scholar
  29. 29.
    Minowada J, Ohnuma T, Moore GE (1972) Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst 49: 891–895Google Scholar
  30. 30.
    Miyoshi I, Taguchi H, Kubonishi I, Yoshimoto S, Ohtsuki Y, Shiraishi Y, Agaki T (1982) Type C virus-producing cell lines derived from adult T cell leukemia. In: Hanaoka M, Takatsuki K, Shimoyama M (ed) Adult T cell leukemia and related diseases. Japan Scientific Societies Press, Tokyo, Plenum Press, New York (GANN Monograph on cancer research, no 28, pp 219–228)Google Scholar
  31. 31.
    Nakai M, Imura S, Goto T, Imagawa DT, Sano K (1986) Ultrastructural features of AIDS virus morphogenesis. J Electron Microsc 35 [Suppl]: 3445–3446Google Scholar
  32. 32.
    Nakashima H, Koyanagi Y, Harada S, Yamamoto N (1986) Effect of HTLV-III on the macromolecular synthesis of HTLV-I carrying cell line, MT-4. Med Microbiol Immunol 175: 325–334Google Scholar
  33. 33.
    Palmer E, Sporborg C, Harrison A, Martin ML, Feorino P (1985) Morphology and immunoelectron microscopy of AIDS virus. Arch Virol 85: 189–196Google Scholar
  34. 34.
    Pinter A, de Harven E (1979) Protein composition of a defective murine sarcoma virus particle possessing the enveloped type-A morphology. Virology 99: 103–110Google Scholar
  35. 35.
    Popovic M, Read-Connole E, Gallo RC (1984) T4 positive human neoplastic cell lines susceptible to and permissive for HTLV-III. Lancet ii: 1472–1473Google Scholar
  36. 36.
    Popovic M, Sarngadharan MG, Read E, Gallo RC (1984) Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science 224: 497–500Google Scholar
  37. 37.
    Sarkar NH, Nowinski RC, Moore DH (1971) Helical nucleocapsid structure of the oncogenic ribonucleic acid viruses (oncornaviruses). J Virol 8: 564–572Google Scholar
  38. 38.
    Simionescu N, Simionescu M (1976) Galloylglucoses of low molecular weight as mordant in electron microscopy I. Procedure, and evidence for mordanting effect. J Cell Biol 70: 608–621Google Scholar
  39. 39.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354Google Scholar
  40. 40.
    Yoshinaka Y, Luftig RB (1977) Murine leukemia virus morphogenesis: Cleavage of P70 in vitro can be accompanied by a shift from a concentrically coiled internal strand (“immature”) to a collapsed (“mature”) form of the virus core. Proc Natl Acad Sci USA 74: 3446–3450Google Scholar
  41. 41.
    Yoshinaka Y, Luftig RB (1977) Properties of a P70 proteolytic factor of murine leukemia viruses. Cell 12: 709–719Google Scholar
  42. 42.
    Yoshinaka Y, Luftig RB (1978) Morphological conversion of “immature” Rauscher leukaemia virus cores to a “mature” form after addition of the P65–70 (gag gene product) proteolytic factor. J Gen Virol 40: 151–160Google Scholar
  43. 43.
    Yoshinaka Y, Luftig RB (1982) p 65 of Gazdar murine sarcoma viruses contains antigenic determinants from all four of the murine leukemia virus (MuLV)gag polypeptides (p 15, p 12, p 30, and p 10) and can be cleaved in vitro by the MuLV proteolytic activity. Virology 118: 380–388Google Scholar
  44. 44.
    Zhang JJ, Goto T, Morita C, Nakai M, Ikuta K, Kato S (1988) Localization of the components of human immunodeficiency virus by immunocolloidal gold labeling. J Clin Electron Microsc 21: 625–626Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • T. Goto
    • 1
  • K. Ikuta
    • 2
  • J. J. Zhang
    • 1
  • C. Morita
    • 1
  • K. Sano
    • 1
  • M. Komatsu
    • 4
  • H. Fujita
    • 4
  • S. Kato
    • 3
  • M. Nakai
    • 1
  1. 1.Department of MicrobiologyOsaka Medical CollegeTakatsuki, Osaka
  2. 2.Division of Serology, Institute of Immunological ScienceHokkaido UniversityKita-ku, Sapporo
  3. 3.Department of Pathology, Research Institute for Microbial DiseasesOsaka UniversitySuita, OsakaJapan
  4. 4.Research Center of UHVEMOsaka UniversitySuita, OsakaJapan

Personalised recommendations