Summary
The expression of the capsid antigen (CA) and the two regulatory proteinsnef andvpu as well as the CD4 cell surface receptor was followed in HIV-infected lymphoid and promonocytic cells. In the lytic phase of infection all three viral proteins were expressed; production of these proteins coincided with the increase of CA antigen and infectious virus in culture supernatants and with prominent cytopathic effects. After selection of persistently infected cells, the number of lymphoid cells expressing detectable levels ofnef decreased to zero; the number of cells positive for CA ranged between 40 to 70%. In chronically infected promonocytic cellsnef andvpu expression was reduced to undetectable levels, whereas most of the cells accumulated CA intracellularly. Infectious cell free virus and CA in the supernatant of promonocytic cells had low titers. CD4 surface expression declined in all cell lines investigated before cell free virus was detectable.
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References
Ahmad N, Venkatesan S (1988)Nef protein is a transcriptional repressor of HIV-1 LTR. Science 241: 1481–1485
Albert J, Bredeberg U, Chiodi F, Böttinger B, Fenyö EM, Norrby E, Biberfeld G (1987) A new human retrovirus of West African origin (SBL6669) and its relationship to HTLV-IV, LAV-II, and HTLV-IIIB. AIDS Res Hum Retroviruses 3: 3–10
Åsjö B, Ivhed I, Gidlund M, Fuerstenberg S, Fenyö EM, Nilsson K, Wigzell H (1987) Susceptibility to infection by the human immunodeficiency virus (HIV) correlates with T4 expression in an parental monocytoid cell line and its subclones. Virology 157: 359–365
Bachelerie F, Alcami J Hazan U, Israël N, Goud B, Arenzana-Seisdedos F, Virelizier J-L (1990) Constitutive expression of human immunodeficiency virus (HIV)nef protein in human astrocytes does not influence basal or induced HIV long terminal repeat activity. J Virol 64: 3059–3062
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: 868–871
Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins form sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated Protein A. Anal Biochem 112: 195–203
Bröker M (1986) Vectors for regulated high-level expression of proteins fused to truncated forms ofEschericha coli β-galactosidase. Gene Anal Techn 3: 53–57
Bruce C, Buonocore L, Rose JK (1990) CD4 is retained in the endoplasmatic reticulum by the human immunodeficiency virus type 1 glycoprotein precursor. J Virol 64: 5585–5593
Cheng-Mayer C, Jannello P, Shaw K, Luciw PA, Levy JA (1989) Differential effects ofnef on HIV replication: implications for viral pathogenesis in the host. Science 246: 1629–1632
Cohen EA, Terwilliger EF, Sodroski JG, Haseltine WA (1988) Identification of a protein encoded by thevpu gene of HIV-1. Nature 334: 532–534
Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z, MacDonald S, Pulford KAF, Stein H, Mason DY (1984) Immunoenzymatic labeling of monoclonal antibodies using complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complex). J Histochem Cytochem 32: 219–229
Cullen BR (1991) The positive effect of the negative factor. Nature 351: 698–699
Daniel MD, Letvin NL, King NW, Kannagi M, Seghal PK, Hunt RD, Kanki PJ, Essex M, Desrosiers RC (1985) Isolation of a T-cell tropic HTLV-III-like retrovirus from macaques. Science 228: 1201–1204
Delassus S, Cheynier R, Wain-Hobson S (1991) Evolution of human immunodeficiency virus type 1nef and long terminal repeat sequences over 4 years in vitro. J Virol 65: 225–231
Durda PJ, Leece B, Jenoski A, Rabin H, Fisher A, Wong-Staal F (1988) Characterization of murine monoclonal antibodies to HIV-1 induced by peptides. AIDS Res Human Retroviruses 4: 331–342
Franchini G, Ropert-Guroff M, Ghrayeb J, Chang NT, Wong-Staal F (1986) Cytoplasmic localization of the HTLV-III 3′-orf protein in cultured T-cells. Virology 155: 593–599
Garcia JV, Miller AD (1991) Serine phosphorylation independent downregulation of cell surface CD4 bynef. Nature 350: 508–511
Gelderblom H, Reupke H, Winkel T, Kunze R, Pauli G (1987) MHC-antigens: constituents of the envelope of human and simian immunodeficiency virus. Z Naturforsch 42: 1328–1334
Guy B, Kieny MP, Riviere Y, Peuch CL, Dott K, Girad M, Montagnier L, Lecocq JP (1987) HIV F/3′-orf encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature 330: 266–269
Guy B, Riviere Y, Dott K, Regnault A, Kieny MP (1990) Mutational analysis of the HIVnef protein. Virology 176:413–425
Hammes S, Dixon EP, Malim MH, Bryan MH, Cullen BR, Greene WC (1989)Nef protein of human imunodeficiency virus type 1: evidence against its role as a transcriptional inhibitor. Proc Natl Acad Sci USA 86: 9549–9553
Jabbar MA, Nayak DP (1990) Intracellular interaction of human immunodeficiency virus type 1 (ARV-2) envelope gylcoprotein gp160 with CD4 blocks the movement and maturation of CD4 to the plasma membrane. J Virol 64: 6297–6304
Kestler HW, Ringler DJ, Kazuyasu M, Panicali DL, Sehgal PK, Daniel MD, Desrosiers RC (1991) Importance of thenef gene for maintenance of high virus loads and for development of AIDS. Cell 65: 651–662
Kikukawa R, Koyanagi Y, Harade S, Kobayashi N, Hatanaka M, Yamamoto N (1986) Differential susceptibility to the acquired immunodeficiency syndrome retrovirus in cloned cells of human leukemic T-cell line MOLT-4. J Virol 57: 1159–1162
Kim S, Ikeuchi K, Byrn R, Groopman J, Baltimore D (1989) Lack of a negative influence on viral growth by thenef gene of human immunodeficiency virus type 1. Proc Natl Acad Sci USA 86: 9544–9548
Klimkait T, Strebel K, Hoggan MD, Martin MA, Orenstein JM (1990) The human immunodeficency virus type 1-specific proteinvpu is required for efficent virus maturation and release. J Virol 64: 621–629
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibodies of predefined specificity. Nature 256: 495–497
Kraus G, Werner A, Baier M, Binninger D, Ferdinand FJ, Norley S, Kurth R (1989) Isolation of human immunodeficiency virus-related simian immunodeficiency viruses from African green monkeys. Proc Natl Acad Sci USA 86: 2892–2896
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–684
Laurent AG, Hovanessian AG, Riviere Y, Krust B, Regnault A, Montagnier L, Findeli A, Kieny MP, Guy B (1990) Production of a non-functionalnef protein in human immunodeficiency virus type 1-infected CEM cells. J Gen Virol 71: 2273–2281
Liu FT, Zinnecker M, Hamaska T, Katz DH (1979) New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry 18: 690–697
Luciw PA, Cheng-Meyer C, Jay JA (1987) Mutational analysis of the human immunodeficiency virus: The orf-B region down-regulates virus replication. Proc Natl Acad Sci USA 84: 1434–1438
Montagnier L, Dauguet C, Axler C, Chamaret S, Gruest J, Nugeyre MT, Rey F, Barré-Sinoussi F, Chermannn JC (1984) A new type of retrovirus isolated from patients presenting with lymphadenopathy and acquired immune deficiency syndrome: structural and antigenic relatedness with equine infectious anaemia virus. Ann Virol 135E: 119–134
Niederman TMJ, Thielan BJ, Ratner L (1989) Human immunodeficiency virus type 1 negative factor is a transcriptional silencer. Proc Natl Acad Sci USA 86: 1128–1132
Niedrig M, Hinkula J, Weigelt W, L'Age-Stehr J, Pauli G, Rosen J, Wahren B (1989) Epitope mapping of monoclonal antibodies against human immunodeficiency virus type 1 structural proteins by using peptides. J Virol 63: 3525–3528
Niedrig M, Rabanus JP, L'Age-Stehr J, Gelderblom H, Pauli G (1988) Monoclonal antibodies directed against human immunodeficiency virus (HIV) gag proteins with specificity for conserved epitopes in HIV-1, HIV-2 and simian immunodeficiency virus. J Gen Virol 69: 2109–2114
Popovic M, Read-Connole E, Gallo RC (1984) T4 positive human neoplastic cell lines susceptible to and permissive for HTLV-III. Lancet 2: 1472–1473
Popovic M, Sarngadharan MG, Read E, Gallo RC (1984) Detection, isolation and continuous production of cytopathic retroviruses (HTLV-III) from patiens with AIDS and pre-AIDS. Science 224: 497–500
Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Hyg 27: 493–497
Rieber EP, Rank G, Wirth S, Wilhelm M, Kopp E, Riethmüller G (1984) Modulation of T-cell functions by monoclonal “pan T-cell” antibodies not directed against the T-cell receptor complex. In: Reinherz EL, Haynes BF, Nadler LM, Bernstein JD (eds) Leucocyte typing II. Springer, Berlin Heidelberg New York, p 233
Schägger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophorese for the separation of proteins in the range from 1 to 100kD. Anal Biochem 166: 368–379
Schneider T, Harthus H, Hildebrandt P, Niedrig M, Bröker M, Weigelt W, Beck A, Pauli G (1991) Epitopes of the HIV-1 negative factor (nef) reactive with murine monoclonal antibodies and human HIV-1 positive sera. AIDS Res Human Retroviruses 7: 37–44
Schneider T, Hildebrandt P, Rönspeck W, Weigelt W, Pauli G (1990) The antibody response to the HIV-1 specific “out” (vpu) protein: identification of an immunodominant epitope and correlation of antibody detectability to clinical stages. AIDS Res Human Retroviruses 6: 943–950
Strebel K, Klimkait T, Martin MA (1988) A novel gene of HIV-1,vpu and its 16-kilodalton product. Science 241: 1221–1223
Sundstrom C, Nilsson K (1976) Establishment and characterization of a human histocytic lymphoma cell line (U937). Int J Cancer 17: 565–577
Terwilliger EF, Cohen EA, Lu Y, Sodroski JG, Haseltine WA (1989) Functional role of human immunodeficiency virus type 1vpu. Proc Natl Acad Sci USA 86: 5163–5167
Terwilliger EF, Sodroski JG, Rosen GA, Halseltine WA (1986) Effects of mutations within the 3′-orf open reading frame region of human T-cell lymphotropic virus type III (HTLV-III/LAV) on replication and cytopathogenicity. J Virol 60: 754–760
Valentin A, Albert J, Fenyö EM, Åsjö B (1990) HIV-1 infection of normal human macrophage cultures: implication for silent infection. Virology 177: 790–794
Ziegler-Heitbrock HWL, Thiel E, Fütterer A, Herzog V, Wirtz A, Rietmüller G (1988) Establishment of a human cell line (MonoMac6) with characteristics of mature monocytes. Int J Cancer 41: 455–461
Zweig M, Samuel KP, Showalter SD, Bladen SV, DuBois GC, Latenberger JA, Hodge DR, Papas TS (1990) Heterogenity ofnef proteins in cells infected with human immunodeficiency virus type 1. Virology 179: 504–507
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Schneider, T., Hildebrandt, P., Rokos, K. et al. Expression ofnef, vpu, CA and CD4 during the infection of lymphoid and monocytic cell lines with HIV-1. Archives of Virology 125, 161–176 (1992). https://doi.org/10.1007/BF01309635
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DOI: https://doi.org/10.1007/BF01309635