IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality

J Immunol Methods. 2019 Feb:465:13-19. doi: 10.1016/j.jim.2018.11.012. Epub 2018 Nov 26.

Abstract

Background: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs).

Methods: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times.

Results: We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.

Keywords: PBMC; Preanalytical; Quality control.

Publication types

  • Clinical Trial
  • Multicenter Study

MeSH terms

  • Adult
  • Aged
  • Calcium-Binding Proteins / biosynthesis*
  • Female
  • Gene Expression Regulation*
  • Humans
  • Interleukin-8 / biosynthesis*
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / metabolism*
  • Male
  • Middle Aged
  • alpha-Mannosidase / biosynthesis*

Substances

  • CXCL8 protein, human
  • Calcium-Binding Proteins
  • Interleukin-8
  • EDEM3 protein, human
  • alpha-Mannosidase