Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

Diana V. Pastrana, Christopher B. Buck, Yuk Ying S Pang, Cynthia D. Thompson, Philip E. Castle, Peter C. FitzGerald, Susanne Krüger Kjaer, Douglas R. Lowy, John T. Schiller

Research output: Contribution to journalArticle

270 Citations (Scopus)

Abstract

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies.

Original languageEnglish (US)
Pages (from-to)205-216
Number of pages12
JournalVirology
Volume321
Issue number2
DOIs
StatePublished - Apr 10 2004
Externally publishedYes

Fingerprint

Alkaline Phosphatase
Bovine papillomavirus 1
Serum
Polyomavirus Transforming Antigens
Human papillomavirus 16
Neutralizing Antibodies
Natural History
Reporter Genes
Codon
Antibody Formation
Transfection
Vaccines
Enzyme-Linked Immunosorbent Assay
Genes

Keywords

  • Antibodies
  • HPV
  • Neutralization
  • Papillomavirus
  • Pseudovirus
  • Serology
  • Vaccine
  • Vector
  • Virion
  • Virus-like particles

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Pastrana, D. V., Buck, C. B., Pang, Y. Y. S., Thompson, C. D., Castle, P. E., FitzGerald, P. C., ... Schiller, J. T. (2004). Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. Virology, 321(2), 205-216. https://doi.org/10.1016/j.virol.2003.12.027

Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. / Pastrana, Diana V.; Buck, Christopher B.; Pang, Yuk Ying S; Thompson, Cynthia D.; Castle, Philip E.; FitzGerald, Peter C.; Kjaer, Susanne Krüger; Lowy, Douglas R.; Schiller, John T.

In: Virology, Vol. 321, No. 2, 10.04.2004, p. 205-216.

Research output: Contribution to journalArticle

Pastrana, DV, Buck, CB, Pang, YYS, Thompson, CD, Castle, PE, FitzGerald, PC, Kjaer, SK, Lowy, DR & Schiller, JT 2004, 'Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18', Virology, vol. 321, no. 2, pp. 205-216. https://doi.org/10.1016/j.virol.2003.12.027
Pastrana, Diana V. ; Buck, Christopher B. ; Pang, Yuk Ying S ; Thompson, Cynthia D. ; Castle, Philip E. ; FitzGerald, Peter C. ; Kjaer, Susanne Krüger ; Lowy, Douglas R. ; Schiller, John T. / Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. In: Virology. 2004 ; Vol. 321, No. 2. pp. 205-216.
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