Baseline levels and temporal stability of 27 multiplexed serum cytokine concentrations in healthy subjects

PLoS One. 2013 Dec 12;8(12):e76091. doi: 10.1371/journal.pone.0076091. eCollection 2013.

Abstract

Background: Cytokines are humoral molecules that elicit regulatory function in immunologic pathways. The level and type of cytokine production has become critical in distinguishing physiologic from pathologic immune conditions. Cytokine profiling has become an important biomarker discovery tool in monitoring of the immune system. However, the variations in cytokine levels in individual subjects over time in healthy individuals have not been extensively studied. In this study, we use multiplex bead arrays to evaluate 27 analytes in paired serum samples taken seven days apart from 144 healthy individuals in order to assess variations over a short time period.

Methods: Fluorescent bead-based immunoassay (Luminex) was used to measure 27 analytes in serum samples. Measurements were performed on matched samples from 144 healthy donors. To assess inter-plate variability, one arbitrarily selected serum sample was analyzed on each of the first ten plates as bridge sample.

Results: Using the bridge sample, we showed minimal inter-plate variations in the measurement of most analytes. In measurement of cytokines from the 144 patients at two time points, we found that three cytokines (IL-2, IL-15 and GM-CSF) were undetectable and five analytes (RANTES, MCP-1, VEGF, MIP-1β and PDGF-BB) showed significant difference in concentrations at Day 0 compared to Day 7.

Conclusions: The current study demonstrated higher variations in cytokine levels among individuals than were observed for samples obtained one week apart from identical donors. These data suggest that a serum sample from each subject for use as a baseline measurement is a better control for clinical trials rather than sera from a paired cohort.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adult
  • Aged
  • Becaplermin
  • Chemokine CCL2 / blood
  • Chemokine CCL4 / blood
  • Chemokine CCL5 / blood
  • Cytokines / blood*
  • Female
  • Granulocyte-Macrophage Colony-Stimulating Factor / blood
  • Humans
  • Immunoassay
  • Interleukin-15 / blood
  • Interleukin-2 / blood
  • Male
  • Middle Aged
  • Proto-Oncogene Proteins c-sis / blood
  • Vascular Endothelial Growth Factor A / blood

Substances

  • Chemokine CCL2
  • Chemokine CCL4
  • Chemokine CCL5
  • Cytokines
  • Interleukin-15
  • Interleukin-2
  • Proto-Oncogene Proteins c-sis
  • Vascular Endothelial Growth Factor A
  • Becaplermin
  • Granulocyte-Macrophage Colony-Stimulating Factor

Grant support

This work was supported by the NIH CHI intramural research program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.