Determination of cytokine responses using a multiplexed fluorescent microsphere immunoassay

We used a multiplexedfluorescent microsphere immunoassay to develop a sandwich capture assay to assess simultaneously the production of thymus helper (TH) 1- and TH2-type cytokines in tissue culture supernatant obtained from stimulated peripheral blood mononuclear cells. The assay then was used to a...

Full description

Saved in:
Bibliographic Details
Published in:American journal of clinical pathology 2002-09, Vol.118 (3), p.346-353
Main Authors: MARTINS, Thomas B, PASI, Brian M, PICKERING, Jerry W, JASKOWSKI, Troy D, LITWIN, Christine M, HILL, Harry R
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cells, Cultured
Cytokines - biosynthesis
Fetal Blood - immunology
Fluorescence
Humans
Hypergammaglobulinemia - immunology
Immunoassay - instrumentation
Immunoassay - methods
Immunoglobulin E
Immunologic Deficiency Syndromes - immunology
Infant, Newborn
Investigative techniques, diagnostic techniques (general aspects)
Leukocytes, Mononuclear - immunology
Lymphocyte Activation - physiology
Medical sciences
Microspheres
Miscellaneous. Technology
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Sensitivity and Specificity
Staphylococcus
Staphylococcus aureus - immunology
Th1 Cells - immunology
Th2 Cells - immunology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We used a multiplexedfluorescent microsphere immunoassay to develop a sandwich capture assay to assess simultaneously the production of thymus helper (TH) 1- and TH2-type cytokines in tissue culture supernatant obtained from stimulated peripheral blood mononuclear cells. The assay then was used to assess the cytokine production of patients with hyperimmunoglobulinemia E syndrome and in cord blood from neonates. The multiplexed assay has a reportable range of less than 10 to 50,000 pg/mL. For linearity and recovery studies, R2 values for the 6 cytokines ranged from 0.988 to 0.999 for samples spiked with known concentrations of recombinant cytokine standards and for patient samples. The assay showed good specificity, with little cross-reactivity between cytokines. Results from supernatants of Staphylococcus aureus-stimulated peripheral blood mononuclear cells obtainedfrom 6 patients with hyperimmunoglobulinemia E syndrome showed significantly less interferon (IFN)-gamma production than cells from healthy control subjects. Cord blood cells from neonates produced significantly less interleukin 12 and IFN-gamma than cells from adults in group B streptococci-stimulated mononuclear cells. The fluorescent multiplexed microsphere immunoassay can be used to quantitate multiple cytokines from 1 sample and should be useful for further understanding of the cytokine role in disease.
ISSN:0002-9173
1943-7722
DOI:10.1309/N0T6-C56B-GXB2-NVFB