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Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay.
J Virol Methods. 1998 Mar; 71(1):35-44.JV

Abstract

A FACS-measured immunofluorescence assay was developed for the detection of antibodies directed against the hemagglutinin (H) and fusion (F) glycoproteins of measles virus (MV). Human melanoma cell lines transfected with either the MV H or F genes, which showed a high surface expression of the respective proteins in their native conformation, were used as target cells. The cells were incubated with diluted plasma samples, and stained subsequently with FITC-conjugated secondary antibodies. The FACS-measured fluorescence signals correlated directly with the amount of specific immunoglobulins over a wide concentration range. The use of different conjugates enabled the separate detection of MV-specific IgG, IgM, IgA and IgG subclasses, with relatively low backgrounds. Hemagglutinin-specific IgG, IgM and IgA fluorescence signals were shown to correlate well with MV-specific IgG ELISA titers and MV-specific IgM or IgA capture ELISA OD450-values, respectively. The polyclonal conjugates with specificity for human immunoglobulins offered sufficient cross-reactivity to detect MV-specific IgG, IgM and IgA in plasma samples of cynomolgus macaques, making this technique a useful tool for studying serological responses in vaccination and challenge experiments in non-human primate models.

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Authors+Show Affiliations

de Swart RL
Institute of Virology, Erasmus University, Rotterdam, The Netherlands. deswart@viro.fgg.eur.nl
Vos HW
No affiliation info available
UytdeHaag FG
No affiliation info available
Osterhaus AD
No affiliation info available
van Binnendijk RS
No affiliation info available

MeSH

AnimalsAntibodies, ViralCell LineEnzyme-Linked Immunosorbent AssayFluorescent Antibody Technique, IndirectHemagglutinins, ViralHumansImmunoglobulin AImmunoglobulin GImmunoglobulin MMacaca fascicularisMeasles virusMelanocytesTransfectionViral Fusion Proteins

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9628219

Citation

de Swart, R L., et al. "Measles Virus Fusion Protein- and Hemagglutinin-transfected Cell Lines Are a Sensitive Tool for the Detection of Specific Antibodies By a FACS-measured Immunofluorescence Assay." Journal of Virological Methods, vol. 71, no. 1, 1998, pp. 35-44.
de Swart RL, Vos HW, UytdeHaag FG, et al. Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay. J Virol Methods. 1998;71(1):35-44.
de Swart, R. L., Vos, H. W., UytdeHaag, F. G., Osterhaus, A. D., & van Binnendijk, R. S. (1998). Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay. Journal of Virological Methods, 71(1), 35-44.
de Swart RL, et al. Measles Virus Fusion Protein- and Hemagglutinin-transfected Cell Lines Are a Sensitive Tool for the Detection of Specific Antibodies By a FACS-measured Immunofluorescence Assay. J Virol Methods. 1998;71(1):35-44. PubMed PMID: 9628219.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay. AU - de Swart,R L, AU - Vos,H W, AU - UytdeHaag,F G, AU - Osterhaus,A D, AU - van Binnendijk,R S, PY - 1998/6/17/pubmed PY - 1998/6/17/medline PY - 1998/6/17/entrez SP - 35 EP - 44 JF - Journal of virological methods JO - J Virol Methods VL - 71 IS - 1 N2 - A FACS-measured immunofluorescence assay was developed for the detection of antibodies directed against the hemagglutinin (H) and fusion (F) glycoproteins of measles virus (MV). Human melanoma cell lines transfected with either the MV H or F genes, which showed a high surface expression of the respective proteins in their native conformation, were used as target cells. The cells were incubated with diluted plasma samples, and stained subsequently with FITC-conjugated secondary antibodies. The FACS-measured fluorescence signals correlated directly with the amount of specific immunoglobulins over a wide concentration range. The use of different conjugates enabled the separate detection of MV-specific IgG, IgM, IgA and IgG subclasses, with relatively low backgrounds. Hemagglutinin-specific IgG, IgM and IgA fluorescence signals were shown to correlate well with MV-specific IgG ELISA titers and MV-specific IgM or IgA capture ELISA OD450-values, respectively. The polyclonal conjugates with specificity for human immunoglobulins offered sufficient cross-reactivity to detect MV-specific IgG, IgM and IgA in plasma samples of cynomolgus macaques, making this technique a useful tool for studying serological responses in vaccination and challenge experiments in non-human primate models. SN - 0166-0934 UR - https://www.unboundmedicine.com/medline/citation/9628219/Measles_virus_fusion_protein__and_hemagglutinin_transfected_cell_lines_are_a_sensitive_tool_for_the_detection_of_specific_antibodies_by_a_FACS_measured_immunofluorescence_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(97)00188-2 DB - PRIME DP - Unbound Medicine ER -