Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis

W.E. Marissen, R.A. Kramer, A. Rice, W.C. Weldon, M. Niezgoda, M. Faber, J.W. Slootstra, R.H. Meloen, M. Clijsters-van der Horst, T.J. Visser, M. Jongeneelen, S. Thijsse, M. Throsby, J. de Kruif, C.E. Rupprecht, B. Dietzschold, J. Goudsmit, A.B.H. Bakker

    Research output: Contribution to journalArticleAcademicpeer-review

    57 Citations (Scopus)

    Abstract

    Anti-rabies virus immunoglobulin combined with rabies vaccine protects humans from lethal rabies infections. For cost and safety reasons, replacement of the human or equine polyclonal immunoglobulin is advocated, and the use of rabies virus-specific monoclonal antibodies (MAbs) is recommended. We produced two previously described potent rabies virus-neutralizing human MAbs, CR57 and CRJB, in human PER.C6 cells. The two MAbs competed for binding to rabies virus glycoprotein. Using CR57 and a set of 15-mer overlapping peptides covering the glycoprotein ectodomain, a neutralization domain was identified between amino acids (aa) 218 and 240. The minimal binding region was identified as KLCGVL (aa 226 to 231), with key residues K-CGV- identified by alanine replacement scanning. The critical binding region of this novel nonconformational rabies virus epitope is highly conserved within rabies viruses of genotype 1. Subsequently, we generated six rabies virus variants escaping neutralization by CR57 and six variants escaping CRJB. The CR57 escape mutants were only partially covered by CRJB, and all CRJB-resistant variants completely escaped neutralization by CR57. Without exception, the CR57-resistant variants showed a mutation at key residues within the defined minimal binding region, while the CRJB escape viruses showed a single mutation distant from the CR57 epitope (N182D) combined with mutations in the CR57 epitope. The competition between CR57 and CRJB, the in vitro escape profile, and the apparent overlap between the recognized epitopes argues against including both CR57 and CRJB in a MAb cocktail aimed at replacing classical immunoglobulin preparations.
    Original languageEnglish
    Pages (from-to)4672-4678
    JournalJournal of Virology
    Volume79
    Issue number8
    DOIs
    Publication statusPublished - 2005

    Fingerprint

    Rabies virus
    neutralization
    epitopes
    Epitopes
    monoclonal antibodies
    Monoclonal Antibodies
    mutants
    immunoglobulins
    Immunoglobulins
    rabies
    mutation
    Mutation
    glycoproteins
    Glycoproteins
    Rabies Vaccines
    Amino Acids
    amino acids
    Rabies
    lethal genes
    Alanine

    Keywords

    • high-level expression
    • postexposure prophylaxis
    • glycoprotein
    • pathogenicity
    • determinant
    • virulence
    • mice
    • igg

    Cite this

    Marissen, W. E., Kramer, R. A., Rice, A., Weldon, W. C., Niezgoda, M., Faber, M., ... Bakker, A. B. H. (2005). Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis. Journal of Virology, 79(8), 4672-4678. https://doi.org/10.1128/JVI.79.8.4672-4678.2005
    Marissen, W.E. ; Kramer, R.A. ; Rice, A. ; Weldon, W.C. ; Niezgoda, M. ; Faber, M. ; Slootstra, J.W. ; Meloen, R.H. ; Clijsters-van der Horst, M. ; Visser, T.J. ; Jongeneelen, M. ; Thijsse, S. ; Throsby, M. ; de Kruif, J. ; Rupprecht, C.E. ; Dietzschold, B. ; Goudsmit, J. ; Bakker, A.B.H. / Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis. In: Journal of Virology. 2005 ; Vol. 79, No. 8. pp. 4672-4678.
    @article{de76751c350f455cb33bc1ace1d12315,
    title = "Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis",
    abstract = "Anti-rabies virus immunoglobulin combined with rabies vaccine protects humans from lethal rabies infections. For cost and safety reasons, replacement of the human or equine polyclonal immunoglobulin is advocated, and the use of rabies virus-specific monoclonal antibodies (MAbs) is recommended. We produced two previously described potent rabies virus-neutralizing human MAbs, CR57 and CRJB, in human PER.C6 cells. The two MAbs competed for binding to rabies virus glycoprotein. Using CR57 and a set of 15-mer overlapping peptides covering the glycoprotein ectodomain, a neutralization domain was identified between amino acids (aa) 218 and 240. The minimal binding region was identified as KLCGVL (aa 226 to 231), with key residues K-CGV- identified by alanine replacement scanning. The critical binding region of this novel nonconformational rabies virus epitope is highly conserved within rabies viruses of genotype 1. Subsequently, we generated six rabies virus variants escaping neutralization by CR57 and six variants escaping CRJB. The CR57 escape mutants were only partially covered by CRJB, and all CRJB-resistant variants completely escaped neutralization by CR57. Without exception, the CR57-resistant variants showed a mutation at key residues within the defined minimal binding region, while the CRJB escape viruses showed a single mutation distant from the CR57 epitope (N182D) combined with mutations in the CR57 epitope. The competition between CR57 and CRJB, the in vitro escape profile, and the apparent overlap between the recognized epitopes argues against including both CR57 and CRJB in a MAb cocktail aimed at replacing classical immunoglobulin preparations.",
    keywords = "high-level expression, postexposure prophylaxis, glycoprotein, pathogenicity, determinant, virulence, mice, igg",
    author = "W.E. Marissen and R.A. Kramer and A. Rice and W.C. Weldon and M. Niezgoda and M. Faber and J.W. Slootstra and R.H. Meloen and {Clijsters-van der Horst}, M. and T.J. Visser and M. Jongeneelen and S. Thijsse and M. Throsby and {de Kruif}, J. and C.E. Rupprecht and B. Dietzschold and J. Goudsmit and A.B.H. Bakker",
    note = "Times Cited: 6",
    year = "2005",
    doi = "10.1128/JVI.79.8.4672-4678.2005",
    language = "English",
    volume = "79",
    pages = "4672--4678",
    journal = "Journal of Virology",
    issn = "0022-538X",
    publisher = "American Society for Microbiology",
    number = "8",

    }

    Marissen, WE, Kramer, RA, Rice, A, Weldon, WC, Niezgoda, M, Faber, M, Slootstra, JW, Meloen, RH, Clijsters-van der Horst, M, Visser, TJ, Jongeneelen, M, Thijsse, S, Throsby, M, de Kruif, J, Rupprecht, CE, Dietzschold, B, Goudsmit, J & Bakker, ABH 2005, 'Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis', Journal of Virology, vol. 79, no. 8, pp. 4672-4678. https://doi.org/10.1128/JVI.79.8.4672-4678.2005

    Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis. / Marissen, W.E.; Kramer, R.A.; Rice, A.; Weldon, W.C.; Niezgoda, M.; Faber, M.; Slootstra, J.W.; Meloen, R.H.; Clijsters-van der Horst, M.; Visser, T.J.; Jongeneelen, M.; Thijsse, S.; Throsby, M.; de Kruif, J.; Rupprecht, C.E.; Dietzschold, B.; Goudsmit, J.; Bakker, A.B.H.

    In: Journal of Virology, Vol. 79, No. 8, 2005, p. 4672-4678.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: Fine mapping and escape mutant analysis

    AU - Marissen, W.E.

    AU - Kramer, R.A.

    AU - Rice, A.

    AU - Weldon, W.C.

    AU - Niezgoda, M.

    AU - Faber, M.

    AU - Slootstra, J.W.

    AU - Meloen, R.H.

    AU - Clijsters-van der Horst, M.

    AU - Visser, T.J.

    AU - Jongeneelen, M.

    AU - Thijsse, S.

    AU - Throsby, M.

    AU - de Kruif, J.

    AU - Rupprecht, C.E.

    AU - Dietzschold, B.

    AU - Goudsmit, J.

    AU - Bakker, A.B.H.

    N1 - Times Cited: 6

    PY - 2005

    Y1 - 2005

    N2 - Anti-rabies virus immunoglobulin combined with rabies vaccine protects humans from lethal rabies infections. For cost and safety reasons, replacement of the human or equine polyclonal immunoglobulin is advocated, and the use of rabies virus-specific monoclonal antibodies (MAbs) is recommended. We produced two previously described potent rabies virus-neutralizing human MAbs, CR57 and CRJB, in human PER.C6 cells. The two MAbs competed for binding to rabies virus glycoprotein. Using CR57 and a set of 15-mer overlapping peptides covering the glycoprotein ectodomain, a neutralization domain was identified between amino acids (aa) 218 and 240. The minimal binding region was identified as KLCGVL (aa 226 to 231), with key residues K-CGV- identified by alanine replacement scanning. The critical binding region of this novel nonconformational rabies virus epitope is highly conserved within rabies viruses of genotype 1. Subsequently, we generated six rabies virus variants escaping neutralization by CR57 and six variants escaping CRJB. The CR57 escape mutants were only partially covered by CRJB, and all CRJB-resistant variants completely escaped neutralization by CR57. Without exception, the CR57-resistant variants showed a mutation at key residues within the defined minimal binding region, while the CRJB escape viruses showed a single mutation distant from the CR57 epitope (N182D) combined with mutations in the CR57 epitope. The competition between CR57 and CRJB, the in vitro escape profile, and the apparent overlap between the recognized epitopes argues against including both CR57 and CRJB in a MAb cocktail aimed at replacing classical immunoglobulin preparations.

    AB - Anti-rabies virus immunoglobulin combined with rabies vaccine protects humans from lethal rabies infections. For cost and safety reasons, replacement of the human or equine polyclonal immunoglobulin is advocated, and the use of rabies virus-specific monoclonal antibodies (MAbs) is recommended. We produced two previously described potent rabies virus-neutralizing human MAbs, CR57 and CRJB, in human PER.C6 cells. The two MAbs competed for binding to rabies virus glycoprotein. Using CR57 and a set of 15-mer overlapping peptides covering the glycoprotein ectodomain, a neutralization domain was identified between amino acids (aa) 218 and 240. The minimal binding region was identified as KLCGVL (aa 226 to 231), with key residues K-CGV- identified by alanine replacement scanning. The critical binding region of this novel nonconformational rabies virus epitope is highly conserved within rabies viruses of genotype 1. Subsequently, we generated six rabies virus variants escaping neutralization by CR57 and six variants escaping CRJB. The CR57 escape mutants were only partially covered by CRJB, and all CRJB-resistant variants completely escaped neutralization by CR57. Without exception, the CR57-resistant variants showed a mutation at key residues within the defined minimal binding region, while the CRJB escape viruses showed a single mutation distant from the CR57 epitope (N182D) combined with mutations in the CR57 epitope. The competition between CR57 and CRJB, the in vitro escape profile, and the apparent overlap between the recognized epitopes argues against including both CR57 and CRJB in a MAb cocktail aimed at replacing classical immunoglobulin preparations.

    KW - high-level expression

    KW - postexposure prophylaxis

    KW - glycoprotein

    KW - pathogenicity

    KW - determinant

    KW - virulence

    KW - mice

    KW - igg

    U2 - 10.1128/JVI.79.8.4672-4678.2005

    DO - 10.1128/JVI.79.8.4672-4678.2005

    M3 - Article

    VL - 79

    SP - 4672

    EP - 4678

    JO - Journal of Virology

    JF - Journal of Virology

    SN - 0022-538X

    IS - 8

    ER -