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

doi: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

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 journal › Article › peer-review

322 Scopus citations

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 language	English (US)
Pages (from-to)	205-216
Number of pages	12
Journal	Virology
Volume	321
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2003.12.027
State	Published - Apr 10 2004
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

Virology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.virol.2003.12.027

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@article{69ec6e0aced24206bd6e9a75f671eef9,

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

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.",

keywords = "Antibodies, HPV, Neutralization, Papillomavirus, Pseudovirus, Serology, Vaccine, Vector, Virion, Virus-like particles",

author = "Pastrana, {Diana V.} and Buck, {Christopher B.} and Pang, {Yuk Ying S.} and Thompson, {Cynthia D.} and Castle, {Philip E.} and FitzGerald, {Peter C.} and Kjaer, {Susanne Kr{\"u}ger} and Lowy, {Douglas R.} and Schiller, {John T.}",

note = "Funding Information: This work was supported by the intramural research program of the National Cancer Institute. We thank R. Roden (The Johns Hopkins University) for his insight on using SEAP as a reporter and P. Day (NCI) for helpful discussions and manuscript revisions.",

year = "2004",

month = apr,

day = "10",

doi = "10.1016/j.virol.2003.12.027",

language = "English (US)",

volume = "321",

pages = "205--216",

journal = "Virology",

issn = "0042-6822",

publisher = "Academic Press Inc.",

number = "2",

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TY - JOUR

T1 - Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

AU - Pastrana, Diana V.

AU - Buck, Christopher B.

AU - Pang, Yuk Ying S.

AU - Thompson, Cynthia D.

AU - Castle, Philip E.

AU - FitzGerald, Peter C.

AU - Kjaer, Susanne Krüger

AU - Lowy, Douglas R.

AU - Schiller, John T.

N1 - Funding Information: This work was supported by the intramural research program of the National Cancer Institute. We thank R. Roden (The Johns Hopkins University) for his insight on using SEAP as a reporter and P. Day (NCI) for helpful discussions and manuscript revisions.

PY - 2004/4/10

Y1 - 2004/4/10

N2 - 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.

AB - 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.

KW - Antibodies

KW - HPV

KW - Neutralization

KW - Papillomavirus

KW - Pseudovirus

KW - Serology

KW - Vaccine

KW - Vector

KW - Virion

KW - Virus-like particles

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U2 - 10.1016/j.virol.2003.12.027

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M3 - Article

C2 - 15051381

AN - SCOPUS:1642446613

SN - 0042-6822

VL - 321

SP - 205

EP - 216

JO - Virology

JF - Virology

IS - 2

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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