Determination of antibody concentration as the main parameter in a dengue virus antibody-dependent enhancement assay using FcγR-expressing BHK cells

Determination of antibody concentration as the main parameter in a dengue virus... Dengue virus (DENV) causes a life-threatening illness, with a wide range of symptoms from mild febrile illness, dengue fever (DF), to life-threatening illness, dengue hemorrhagic fever (DHF). Antibody-dependent enhancement (ADE) is considered to be a risk factor for DHF. In the present study, we determined the parameters for ADE assays using FcγR-expressing BHK cells. Monoclonal antibodies and human serum samples were used in the assays. We examined antibody concentration and virus concentration and analyzed whether antibody concentration or DENV-antibody ratio determines ADE activity. Virus growth was quantified by a conventional plaque titration method using FcγR-expressing BHK cells. The assay allowed the detection of DENV growth with inoculation doses ranging from 10 2 PFU/ml to 10 6 PFU/ml using monoclonal antibodies and undiluted or diluted serum samples. The results indicate that antibody concentration rather than DENV-antibody ratio determines the demonstration of ADE activity. Thus, antibody concentration rather than multiplicity of infection was defined as the main determinant in ADE assays using FcγR-expressing BHK cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Determination of antibody concentration as the main parameter in a dengue virus antibody-dependent enhancement assay using FcγR-expressing BHK cells

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Publisher
Springer Vienna
Copyright
Copyright © 2014 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-013-1787-3
Publisher site
See Article on Publisher Site

Abstract

Dengue virus (DENV) causes a life-threatening illness, with a wide range of symptoms from mild febrile illness, dengue fever (DF), to life-threatening illness, dengue hemorrhagic fever (DHF). Antibody-dependent enhancement (ADE) is considered to be a risk factor for DHF. In the present study, we determined the parameters for ADE assays using FcγR-expressing BHK cells. Monoclonal antibodies and human serum samples were used in the assays. We examined antibody concentration and virus concentration and analyzed whether antibody concentration or DENV-antibody ratio determines ADE activity. Virus growth was quantified by a conventional plaque titration method using FcγR-expressing BHK cells. The assay allowed the detection of DENV growth with inoculation doses ranging from 10 2 PFU/ml to 10 6 PFU/ml using monoclonal antibodies and undiluted or diluted serum samples. The results indicate that antibody concentration rather than DENV-antibody ratio determines the demonstration of ADE activity. Thus, antibody concentration rather than multiplicity of infection was defined as the main determinant in ADE assays using FcγR-expressing BHK cells.

Journal

Archives of VirologySpringer Journals

Published: Jan 1, 2014

References

  • Secondary heterologous dengue infection risk: Disequilibrium between immune regulation and inflammation?
    Sierra, B; Perez, AB; Vogt, K; Garcia, G; Schmolke, K; Aguirre, E; Alvarez, M; Kern, F; Kourí, G; Volk, HD; Guzman, MG
  • Immune response to dengue virus and prospects for a vaccine
    Murphy, BR; Whitehead, SS
  • An immunocytometric assay based on dengue infection via DC-SIGN permits rapid measurement of anti-dengue neutralizing antibodies
    Martin, NC; Pardo, J; Simmons, M; Tjaden, JA; Widjaja, S; Marovich, MA; Sun, W; Porter, KR; Burgess, TH
  • Study of the distribution of antibody-dependent enhancement determinants on dengue 2 isolates using dengue 2-derived monoclonal antibodies
    Morens, DM; Larsen, LK; Halstead, SB
  • Phylogenetic analysis of dengue viruses isolated from imported dengue patients: possible aid for determining the countries where infections occurred
    Ito, M; Yamada, K; Takasaki, T; Pandey, B; Nerome, R; Tajima, S; Morita, K; Kurane, I
  • A simple assay system for infection-enhancing and -neutralizing antibodies to dengue type 2 virus using layers of semi-adherent K562 cells
    Konishi, E; Tabuchi, Y; Yamanaka, A

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