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Fine mapping of HIV-1 Nef-epitopes by monoclonal antibodies

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Summary

A panel of newly isolated murine monoclonal antibodies is described which are specific for the Nef protein of the human immunodeficiency virus type 1 (HIV-1). Epitope mapping using recombinant Nef-related proteins, synthetic peptides and lipopeptides showed 3 independent antigenic determinants located within the regions of amino acids 83–93, 175–190 and 86–166 of the Nef protein. None of the monoclonal antibodies reacted with recombinant Nef proteins of HIV-2.

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References

  1. Ahmad N, Venkatesan S (1988) Nef protein of HIV-1 is a transcriptional repressor of HIV-1 LTR. Science 241: 1481–1485

    Google Scholar 

  2. Allan SJ, Coligan JE, Lee TH, McLane MF, Kauki PJ, Groopman JE, Essex M (1986) A new HTLVIII/LAV encoded antigen detected by antibodies from AIDS patients. Science 230: 810–813

    Google Scholar 

  3. Ameisen JC, Guy B, Chamaret S, Loche M, Mouton Y, Neyrinck JL, Khalife J, Leprevost C, Beaucaire G, Boutillon C, Grasse-Masse H, Manziez M, Kieny MP, Laustriat D, Berthier A, Mach B, Montagnier L, Lecocq JP, Capron A (1989) Antibodies to the Nef protein and to Nef peptides in HIV-1 infected seronegative individuals. AIDS 5: 279–291

    Google Scholar 

  4. Beck-Sickinger AG, Dürr H, Jung G (1991) Semiautomated T-bag peptide synthesis using 9-fluorenyl-methoxycarbonyl-strategy and benzotriazol-1-yl-tetramethyl uronium tetrafluoroborate activation. Peptide Res 4: 88–94

    Google Scholar 

  5. Böltz T, Hummel R-P, Tröger W, Rübsamen-Waigmann H, Biesert L, Müller-Lantzsch N, Koch P, Bessler W, Jung G (1988) Distinction between HIV-1 and HIV-2 infection using novel synthetic lipopeptide conjugates as antigens in enzyme immunoassays. J Virol Meth 22: 173–182

    Google Scholar 

  6. Brack-Werner R, Kleinschmidt A, Ludvigsen A, Mellert W, Neumann M, Herrmann R, Khim MCL, Burny A, Müller-Lantzsch N, Stavrou D, Erfle V (1992) Infection of human brain cells by human immunodeficiency virus-1 (HIV-1): restricted virus production in chronically infected human glial cell lines. AIDS 6: 273–285

    Google Scholar 

  7. Bröker M (1986) Vectors for regulated high-level expression of proteins fused to truncated forms ofEscherichia coli β-galactosidase. Gene Anal Techn 3: 53–57

    Google Scholar 

  8. Bröker M, Harthus HP (1989) Defined monoclonal antibodies toEscherichia coli β-galactosidase as a tool for characterization of recombinant expression products. FEBS Lett 257: 118–122

    Google Scholar 

  9. Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112: 195–203

    Google Scholar 

  10. Cheingsong-Popov R, Panagiotidi C, Munaf A, Bowcock S, Watkins P, Aronstam A, Wassef M, Weber J (1990) Prevalence, significance and relationship to seroconversion. AIDS Res Human Retrovir 6: 1099–1105

    Google Scholar 

  11. Chenciner N, Michel F, Dadaglio G, Langlade-Demoyen P, Hoffenbach A, Leroux A, Garcia-Pons F, Rautmann G, Guy B, Guillon JM (1989) Multiple subsets of HIV-specific cytotoxic T lymphocytes in humans and in mice. Eur J Immunol 19: 1537–1544

    Google Scholar 

  12. Cullen BR, Green WC (1990) Functions of the auxiliary gene products of the human immunodeficiency virus type 1. Virology 178: 1–5

    Google Scholar 

  13. Culmann B, Gomard E, Kieny MP, Guy B, Dreyfus F, Saimot AG, Sereni D, Sicard D, Levy JP (1991) Six epitopes reacting with human cytotoxic CD 8+ T cells in the central region of the HIV-1 Nef protein. J Immunol 146: 1560–1565

    Google Scholar 

  14. De Santis R, Anastasi AM, Marcolini S, Valesini G, Pezella M, Vonesch N, Sturchio E, Mele A (1991) Production of a Nef-specific monoclonal antibody by the use of a synthetic peptide. AIDS Res Human Retrovir 7: 315–321

    Google Scholar 

  15. Franchini G, Robert-Guroff M, Ghrayeb J, Chang NT, and Wong-Staal F (1986) Cytoplasmic localization of the HTLV-III 3′ orf protein in cultured T cells. Virology 155: 593–599

    Google Scholar 

  16. Franchini G, Robert-Guroff M, Wong-Staal F, Ghrayeb J, Kato I, Chang TW (1986) Expression of the protein encoded by the 3′ open reading frame of human T cell lymphotropic virus type III in bacteria: demonstration of its immunoreactivity with human sera. Proc Natl Acad Sci USA 83: 5282–5285

    Google Scholar 

  17. Garcia JV, Miller AD (1991) Serine phosphorylation-independent downregulation of cell surface CD 4 by Nef. Nature 350: 508–511

    Google Scholar 

  18. Geysen HM, Meloen RH, Barteling S (1984) Use of peptide synthesis to probe viral antigens to a resolution of a single amino acid. Proc Natl Acad Sci USA 81: 39998–40002

    Google Scholar 

  19. Gombert FO, Blecha W, Tähtinen M, Ranki A, Pfeifer S, Tröger W, Müller-Lantzsch N, Jung G, Rübsamen-Waigmann H, Krohn K (1990) Antigenic epitopes of Nef proteins from different HIV-1 strains as recognized by sera from patients with manifest and latent HIV infection. Virology 176: 458–466

    Google Scholar 

  20. Guy B, Kieny MP, Reviere Y, LePeuch C, Dott K, Girard M, Montagnier L, Lecocq JP (1987) HIVF/3′ orf encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature 330: 266–269

    Google Scholar 

  21. Guy B, Riviere J, Dott K, Regnault A, Kieny MP (1990) Mutational analysis of the Nef protein. Virology 176: 413–425

    Google Scholar 

  22. Hammes SR, Dixon EP, Malim MH, Cullen BR, Green WC (1989) Nef protein of human immundeficiency virus type 1: evidence against its role as a transcriptional inhibitor. Proc Natl Acad Sci USA 86: 9549–9553

    Google Scholar 

  23. Jung G, Wiesmüller K-H, Becker G, Bühring H-J, Bessler WG (1985) Increased production of specific antibodies by presentation of the antigen determinants with covalently coupled lipopeptide mitogens. Angew Chem (Int Ed Engl) 24: 872–873

    Google Scholar 

  24. Kaminchik J, Bashan N, Itach A, Sarver N, Gorecki M, Panet A (1991) Genetic characterization of human immunodeficiency virus type 1 Nef gene products translated in vitro and expressed in mammalian cells. J Virol 65: 583–588

    Google Scholar 

  25. Kearney JF, Radbruch A, Liesegang B, Rajewsky K (1979) A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 123: 1548–1550

    Google Scholar 

  26. Kestler HW, Ringler DJ, Mori K, 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

    Google Scholar 

  27. Kienzle N, Bröker M, Harthus HP, Enders M, Erfle V, Buck M, Müller-Lantzsch N (1991) Immunological study of the Nef protein from HIV-1 by polyclonal and monoclonal antibodies. Arch Virol 118: 29–41

    Google Scholar 

  28. Kienzle N, Enders M, Buck M, Siakkou H, Jahn S, Petzold G, Schneweis KE, Bachmann M, Müller WEG, Müller-Lantzsch N (1992) Expression of the HIV-1 Nef protein in the baculovirus system: investigation of anti-Nef antibodies response in human sera and subcellular localization of Nef. Arch Virol 126: 293–301

    Google Scholar 

  29. Kim S, Ikeuchi K, Byrn R, Groopman J, Baltimore D (1989) Lack of a negative influence on viral growth by the Nef gene of human immunodeficiency virus type 1. Proc Natl Acad Sci USA 86: 9544–9548

    Google Scholar 

  30. Kohleisen B, Neumann M, Herrmann R, Brack-Werner R, Krohn KJE, Ovod V, Ranki A, Erfle V (1992) Cellular localization of Nef expressed in persistently HIV-1 infected low-producer astrocytes. AIDS (in press)

  31. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  32. Metzger J, Wiesmüller K-H, Schaude R, Bessler WG, Jung G (1991) Synthesis of novel immunologically active tripalmitoyl-S-glycerylcysteinyl lipopeptides as useful intermediates for immunogen preparations. Int J Peptide Protein Res 37: 46–57

    Google Scholar 

  33. Muesing MA, Smith DH, Cabradilla CD, Benton CV, Lasky LA, Capon DJ (1985) Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. Nature 313: 450–458

    Google Scholar 

  34. Myers G, Rabson AB, Smith TF, Wong-Staal F, Berzofsky JA (eds) (1990) A compilation and analysis of nucleic acid and amino acid sequences. Human Retroviruses and AIDS 1990, Los Alamos National Laboratory

  35. 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

    Google Scholar 

  36. Porstmann T, Wietschke R, Grunow R, Jahn S, Porstmann B, Schmechta H, Pergande M, von Baehr R (1990) Production and characterization of monoclonal antibodies against human Cu/Zn superoxide dismutase and the establishment of a super-rapid enzyme-linked immunosorbant assay. J Immunol Methods 127: 1–10

    Google Scholar 

  37. Ranki A, Krohn M, Allain JP, Franchini G, Valle SL, Antonen J, Leuther M, Krohn K (1987) Long latency preceeds over seroconversion in sexually transmitted human immunodeficiency virus infection. Lancet ii: 589–593

    Google Scholar 

  38. Ranki A, Järvinen K, Valle SL, Nurmilaakso P, Krohn K (1990) Antibodies to recombinant HIV-1 Nef detected in HIV-1 infection as well as in nonrisk individuals. J AIDS 3: 348–355

    Google Scholar 

  39. Ratner L, Haseltine W, Patarca R, Livak KJ, Starcich B, Josephs SF, Doran ER, Rafalski JA, Whitehorn EA, Baumeister K, Ivanoff L, Petteway SR, Pearson ML, Lautenberger JA, Papas TS, Ghrayeb J, Chang NT, Gallo RC, Wong-Staal F (1985) Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature 313: 277–284

    Google Scholar 

  40. Reiss P, de Ronde A, Lange JMA, de Wolf F, Dekker J, Debouk C, Goudsmit J (1989) Antibody response to the viral negative factor (Nef) in HIV-1 infection: a correlate of levels of HIV-1 expression. AIDS 3: 53–57

    Google Scholar 

  41. Schneider T, Harthus HP, 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 Retrovir 7: 37–44

    Google Scholar 

  42. Schneider T, Hildebrandt P, Rokos K, Schubert U, Rönspeck W, Grund C, Beck A, Blesken R, Kulins G, Oldenburg H, Pauli G (1992) Expression ofnef, vpu, CA and CD 4 during the infection of lymphoid and monocytic cell lines with HIV-1. Arch Virol 125: 161–176

    Google Scholar 

  43. Siakkou H, Ulrich R, Uelze A, Moehring R, Rosenthal S (1990) Immunological characterization of BLV proteins synthesized inE. coli. Acta Virol 34: 256–262

    Google Scholar 

  44. Spindler KR, Rosser DSE, Berk AJ (1984) Analysis of adenovirus transforming proteins from early regions 1 A and 1 B with antisera to inducible fusion antigens produced inEscherichia coli. J Virol 49: 132–141

    Google Scholar 

  45. Spohn R, Gombert FO, Jung G (1992) B-cell epitopes of the Nef protein. Res Virol 143: 70–72

    Google Scholar 

  46. Stanley KK, Luzio JP (1984) Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J 3: 1429–1434

    Google Scholar 

  47. Ulrich R, Siakkou H, Mayer J, Kienzle N, Müller-Lantzsch N, Krüger DH (1992) Subcloning of HIV-2nef genes inE. coli and immunological reactivity of expressed fusion proteins. Virus Genes (in press)

  48. Wain-Hobson S, Sonigo P, Danos O, Cole S, Alizon M (1985) Nucleotide sequence of the AIDS virus, LAV. Cell 40: 9–17

    Google Scholar 

  49. Wiesmüller K-H, Bessler WG, Jung G (1983) Synthesis of the mitogenic S-[2, 3-bis(palmitoyloxy)propyl]-N-palmitoylpentapeptide fromEscherichia coli lipoprotein. Hoppe-Seylers Z Physiol Chem 364: 593–606

    Google Scholar 

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Authors and Affiliations

  1. Institute für Virologie der Medizinischen Fakultät (Charité) der Humboldt-Universität, Berlin

    H. Siakkou, R. Ulrich, C. Grötzinger & D. H. Krüger

  2. Institute für Immunologie der Medizinischen Fakultät (Charité) der Humboldt-Universität, Berlin

    S. Jahn

  3. Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene der Universität des Saarlandes, Homburg

    N. Kienzle & N. Müller-Lantzsch

  4. Robert-Koch-Institut, AIDS-Zentrum des Bundesgesundheitsamtes, Berlin

    T. Schneider & G. Pauli

  5. Institut für Molekulare Virologie, GSF-Forschungszentrum für Umwelt und Gesundheit, Neuherberg

    B. Kohleisen

  6. Institut für Organische Chemie der Universität Tübingen, Tübingen, Germany

    R. Spohn & G. Jung

Authors
  1. H. Siakkou
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  2. S. Jahn
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  3. N. Kienzle
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  4. R. Ulrich
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  5. C. Grötzinger
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  6. T. Schneider
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  7. B. Kohleisen
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  8. G. Pauli
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  9. R. Spohn
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  10. G. Jung
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  11. N. Müller-Lantzsch
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  12. D. H. Krüger
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Siakkou, H., Jahn, S., Kienzle, N. et al. Fine mapping of HIV-1 Nef-epitopes by monoclonal antibodies. Archives of Virology 128, 81–95 (1993). https://doi.org/10.1007/BF01309790

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  • DOI: https://doi.org/10.1007/BF01309790

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