Identification of the key differential transcriptional responses of human whole blood following TLR2 or TLR4 ligation in-vitro

Simon Blankley, Christine M. Graham, Ashleigh Howes, Chloe I. Bloom, Matthew P R Berry, Damien J. Chaussabel, Virginia Pascual, Jacques Banchereau, Marc Lipman, Anne O'Garra

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The use of human whole blood for transcriptomic analysis has potential advantages over the use of isolated immune cells for studying the transcriptional response to pathogens and their products. Whole blood stimulation can be carried out in a laboratory without the expertise or equipment to isolate immune cells from blood, with the added advantage of being able to undertake experiments using very small volumes of blood. Toll like receptors (TLRs) are a family of pattern recognition receptors which recognise highly conserved microbial products. Using the TLR2 ligand (Pam3CSK4) and the TLR4 ligand (LPS), human whole blood was stimulated for 0, 1, 3, 6, 12 or 24 hours at which times mRNA was isolated and a comparative microarray was undertaken. A common NFκB transcriptional programme was identified following both TLR2 and TLR4 ligation which peaked at between 3 to 6 hours including upregulation of many of the NFκB family members. In contrast an interferon transcriptional response was observed following TLR4 but not TLR2 ligation as early as 1 hour post stimulation and peaking at 6 hours. These results recapitulate the findings observed in previously published studies using isolated murine and human myeloid cells indicating that in vitro stimulated human whole blood can be used to interrogate the early transcriptional kinetic response of innate cells to TLR ligands. Our study demonstrates that a transcriptomic analysis of mRNA isolated from human whole blood can delineate both the temporal response and the key transcriptional differences following TLR2 and TLR4 ligation.

Original languageEnglish
Article numbere97702
JournalPLoS One
Volume9
Issue number5
DOIs
Publication statusPublished - 19 May 2014
Externally publishedYes

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taxonomic keys
Ligation
Blood
blood
Toll-Like Receptors
Ligands
transcriptomics
Pattern Recognition Receptors
Messenger RNA
cells
Myeloid Cells
Blood Volume
blood volume
Interferons
interferons
Blood Cells
Up-Regulation
Pathogens
Microarrays
In Vitro Techniques

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Identification of the key differential transcriptional responses of human whole blood following TLR2 or TLR4 ligation in-vitro. / Blankley, Simon; Graham, Christine M.; Howes, Ashleigh; Bloom, Chloe I.; Berry, Matthew P R; Chaussabel, Damien J.; Pascual, Virginia; Banchereau, Jacques; Lipman, Marc; O'Garra, Anne.

In: PLoS One, Vol. 9, No. 5, e97702, 19.05.2014.

Research output: Contribution to journalArticle

Blankley, S, Graham, CM, Howes, A, Bloom, CI, Berry, MPR, Chaussabel, DJ, Pascual, V, Banchereau, J, Lipman, M & O'Garra, A 2014, 'Identification of the key differential transcriptional responses of human whole blood following TLR2 or TLR4 ligation in-vitro', PLoS One, vol. 9, no. 5, e97702. https://doi.org/10.1371/journal.pone.0097702
Blankley, Simon ; Graham, Christine M. ; Howes, Ashleigh ; Bloom, Chloe I. ; Berry, Matthew P R ; Chaussabel, Damien J. ; Pascual, Virginia ; Banchereau, Jacques ; Lipman, Marc ; O'Garra, Anne. / Identification of the key differential transcriptional responses of human whole blood following TLR2 or TLR4 ligation in-vitro. In: PLoS One. 2014 ; Vol. 9, No. 5.
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