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

Olga Kofanova, Camille Bellora, Rocio Aguilar Quesada, Alexandre Bulla, Sonia Panadero-Fajardo, Marc Keipes, Kathi Shea, Mars Stone, Pierre Lescuyer, Fay Betsou

Journal: Journal of immunological methods 2020;465():13-19

PMID: 30496732

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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.

Address: Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address: [email protected].; Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg. Electronic address: [email protected].; Biobanco del Sistema Sanitario Público de Andalucía (SSPA Biobank), Avda. del Conocimiento s/n 18016, Granada, Spain; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address: [email protected].; Service de Médecine de Laboratoire, Hôpitaux Universitaires de Genève, 1211 Genève 14, Switzerland; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada.; Biobanco del Sistema Sanitario Público de Andalucía (SSPA Biobank), Avda. del Conocimiento s/n 18016, Granada, Spain. Electronic address: [email protected].; ZithaGesondheetsZentrum, 36 rue Zithe, 2763, Luxembourg. Electronic address: [email protected].; Precision for Medicine, 8425 Progress Drive, Frederick, MD 21701, USA; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address: [email protected].; Blood Systems Research Institute, 270 Masonic Ave., San Francisco, CA 94118, USA; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address: [email protected].; Service de Médecine de Laboratoire, Hôpitaux Universitaires de Genève, 1211 Genève 14, Switzerland. Electronic address: [email protected].; Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address: [email protected].

DOI: 10.1016/j.jim.2018.11.012

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MeSH Terms: Adult, Aged, Calcium-Binding Proteins, Female, Gene Expression Regulation, Humans, Interleukin-8, Leukocytes, Mononuclear, Male, Middle Aged, alpha-Mannosidase