Development of an opsonophagocytic killing assay using HL-60 cells for detection of functional antibodies against Streptococcus pyogenes

Sanaz Salehi, Claudia M. Hohn, Thomas A. Penfound, James Dale

Research output: Contribution to journalArticle

Abstract

The clinical development of group A streptococcal (GAS) vaccines will require the implementation of a standardized, high-throughput assay to measure the activity of functional opsonic antibodies in vaccine recipients. In the present study, we adapted and modified the HL-60-based protocol that was developed for the detection of opsonic antibodies against Streptococcus pneumoniae for use with multiple M types of GAS. Modifications of the assay conditions permitted the evaluation of 21 different M types of GAS in the assay. The specificity of the antibodymediated opsonization was demonstrated by inhibition with homologous, but not heterologous, M proteins. Maximum rates of opsonophagocytic killing (OPK) of 14 different M types promoted by rabbit antiserum against the 30-valent M proteinbased vaccine were comparable in whole-blood and HL-60 assays. Data are also presented showing OPK serum titers (opsonic index) of naturally acquired human antibodies present in IVIG [intravenous immune globulin (human)]. Results of the HL-60 assay performed on different days using 21 different M types of GAS and IVIG as the antibody source were significantly concordant. This report indicates that the OPK assay conditions may be optimized for the measurement of opsonic antibodies against a number of epidemiologically important M types of GAS and, once standardized, should facilitate the clinical development of effective vaccines to prevent these infections.

Original languageEnglish (US)
Article numbere00617-18
JournalmSphere
Volume3
Issue number6
DOIs
StatePublished - Nov 1 2018

Fingerprint

Streptococcus pyogenes
HL-60 Cells
Intravenous Immunoglobulins
Antibodies
Vaccines
Streptococcal Vaccines
Streptococcus pneumoniae
Immune Sera
Rabbits
Infection
Serum
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Development of an opsonophagocytic killing assay using HL-60 cells for detection of functional antibodies against Streptococcus pyogenes. / Salehi, Sanaz; Hohn, Claudia M.; Penfound, Thomas A.; Dale, James.

In: mSphere, Vol. 3, No. 6, e00617-18, 01.11.2018.

Research output: Contribution to journalArticle

@article{3ffb58d8390c43b0b81332d986ad38f5,
title = "Development of an opsonophagocytic killing assay using HL-60 cells for detection of functional antibodies against Streptococcus pyogenes",
abstract = "The clinical development of group A streptococcal (GAS) vaccines will require the implementation of a standardized, high-throughput assay to measure the activity of functional opsonic antibodies in vaccine recipients. In the present study, we adapted and modified the HL-60-based protocol that was developed for the detection of opsonic antibodies against Streptococcus pneumoniae for use with multiple M types of GAS. Modifications of the assay conditions permitted the evaluation of 21 different M types of GAS in the assay. The specificity of the antibodymediated opsonization was demonstrated by inhibition with homologous, but not heterologous, M proteins. Maximum rates of opsonophagocytic killing (OPK) of 14 different M types promoted by rabbit antiserum against the 30-valent M proteinbased vaccine were comparable in whole-blood and HL-60 assays. Data are also presented showing OPK serum titers (opsonic index) of naturally acquired human antibodies present in IVIG [intravenous immune globulin (human)]. Results of the HL-60 assay performed on different days using 21 different M types of GAS and IVIG as the antibody source were significantly concordant. This report indicates that the OPK assay conditions may be optimized for the measurement of opsonic antibodies against a number of epidemiologically important M types of GAS and, once standardized, should facilitate the clinical development of effective vaccines to prevent these infections.",
author = "Sanaz Salehi and Hohn, {Claudia M.} and Penfound, {Thomas A.} and James Dale",
year = "2018",
month = "11",
day = "1",
doi = "10.1128/mSphereDirect.00617-18",
language = "English (US)",
volume = "3",
journal = "mSphere",
issn = "2379-5042",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - Development of an opsonophagocytic killing assay using HL-60 cells for detection of functional antibodies against Streptococcus pyogenes

AU - Salehi, Sanaz

AU - Hohn, Claudia M.

AU - Penfound, Thomas A.

AU - Dale, James

PY - 2018/11/1

Y1 - 2018/11/1

N2 - The clinical development of group A streptococcal (GAS) vaccines will require the implementation of a standardized, high-throughput assay to measure the activity of functional opsonic antibodies in vaccine recipients. In the present study, we adapted and modified the HL-60-based protocol that was developed for the detection of opsonic antibodies against Streptococcus pneumoniae for use with multiple M types of GAS. Modifications of the assay conditions permitted the evaluation of 21 different M types of GAS in the assay. The specificity of the antibodymediated opsonization was demonstrated by inhibition with homologous, but not heterologous, M proteins. Maximum rates of opsonophagocytic killing (OPK) of 14 different M types promoted by rabbit antiserum against the 30-valent M proteinbased vaccine were comparable in whole-blood and HL-60 assays. Data are also presented showing OPK serum titers (opsonic index) of naturally acquired human antibodies present in IVIG [intravenous immune globulin (human)]. Results of the HL-60 assay performed on different days using 21 different M types of GAS and IVIG as the antibody source were significantly concordant. This report indicates that the OPK assay conditions may be optimized for the measurement of opsonic antibodies against a number of epidemiologically important M types of GAS and, once standardized, should facilitate the clinical development of effective vaccines to prevent these infections.

AB - The clinical development of group A streptococcal (GAS) vaccines will require the implementation of a standardized, high-throughput assay to measure the activity of functional opsonic antibodies in vaccine recipients. In the present study, we adapted and modified the HL-60-based protocol that was developed for the detection of opsonic antibodies against Streptococcus pneumoniae for use with multiple M types of GAS. Modifications of the assay conditions permitted the evaluation of 21 different M types of GAS in the assay. The specificity of the antibodymediated opsonization was demonstrated by inhibition with homologous, but not heterologous, M proteins. Maximum rates of opsonophagocytic killing (OPK) of 14 different M types promoted by rabbit antiserum against the 30-valent M proteinbased vaccine were comparable in whole-blood and HL-60 assays. Data are also presented showing OPK serum titers (opsonic index) of naturally acquired human antibodies present in IVIG [intravenous immune globulin (human)]. Results of the HL-60 assay performed on different days using 21 different M types of GAS and IVIG as the antibody source were significantly concordant. This report indicates that the OPK assay conditions may be optimized for the measurement of opsonic antibodies against a number of epidemiologically important M types of GAS and, once standardized, should facilitate the clinical development of effective vaccines to prevent these infections.

UR - http://www.scopus.com/inward/record.url?scp=85058873063&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058873063&partnerID=8YFLogxK

U2 - 10.1128/mSphereDirect.00617-18

DO - 10.1128/mSphereDirect.00617-18

M3 - Article

VL - 3

JO - mSphere

JF - mSphere

SN - 2379-5042

IS - 6

M1 - e00617-18

ER -