title: Multiplexed measurements of immunomodulator levels in peripheral blood of healthy subjects: Effects of analytical variables based on anticoagulants, age, and gender.
abstract: Multiplex microbead immunoassay (MMIA) is a powerful technology for a wide range of biomedical and clinical applications. It is important to study the normal concentration ranges of immunomodulators under different sample preparation conditions and age groups of subjects in order to more precisely determine their reference values for use in assessing alterations of their levels in disease. The aim of this study was to determine the plasma concentrations of immunomodulators (cytokines, chemokines, and growth factors) in the peripheral blood from healthy subjects by the use of a large multiplex panel, and to determine the effects of different anticoagulants, age, and gender on the immunomodulator levels. In addition, the assay precision for these biomarker analytes was determined. Plasma samples from 107 healthy subjects, aged 18 to 85 years, were collected in three different anticoagulants (sodium citrate, EDTA, Heparin); corresponding serum samples were also obtained. Multiplex microbead immunoassays were performed for measuring a total of 23 analytes including chemokines, cytokines, and growth factors (IL-1β, IL-1ra, IL-2, IL-6, IL-7, IL-8, IL-12 p70, IL-17, IFN-γ, IP-10, MCP-1, PDGF-BB, RANTES, TNF-α, IL-1a, IL-16, HGF, MIG, TNF-β, PDGF-ABBB, EGF, Flt-3 Ligand, VEGF). For these analytes, our results showed that the anticoagulant affected the concentration measurements and the coefficients of variation. However, the relative levels of the analytes (profiles) of samples collected in a particular anticoagulant are consistent. The analytes IL-1β, IL-7, Flt-3 Ligand, and IL-12p70 show the largest variation (up to fourfold) between the age groups. In addition, no statistically significant differences in the level of the analytes were found between the sexes.
authors.affiliation: Department of Chemistry, California State University, Fresno, California, 93740; Center for Comparative Medicine, University of California Davis, Davis, California, 95616; Department of Pathology and Laboratory Medicine, University of California School of Medicine, Davis, California, 95616.
authors.forename: V V
authors.initials: VV
authors.lastname: Krishnan
authors.forename: Resmi
authors.initials: R
authors.lastname: Ravindran
authors.forename: Ted
authors.initials: T
authors.lastname: Wun
authors.forename: Paul A
authors.initials: PA
authors.lastname: Luciw
authors.forename: Imran H
authors.initials: IH
authors.lastname: Khan
authors.forename: Kim
authors.initials: K
authors.lastname: Janatpour
mesh_terms: D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D000925:Anticoagulants / Q000494:pharmacology*; D015415:Biomarkers / Q000097:blood; D018925:Chemokines / Q000097:blood; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007155:Immunologic Factors / Q000097:blood*; D036341:Intercellular Signaling Peptides and Proteins / Q000097:blood; D008297:Male; D008875:Middle Aged; D012737:Sex Factors; D055815:Young Adult
chemical_list: D000925:Anticoagulants; D015415:Biomarkers; D018925:Chemokines; D007155:Immunologic Factors; D036341:Intercellular Signaling Peptides and Proteins
keywords: anticoagulant; chemokines; cytokines; luminex; multiplex microbead; normal range