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Macrophage response to LPS

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Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:TET
Link to TET:FILE
Sample providers :David Hume, Kenneth Baillie
Germ layer:mesoderm
Primary cells or cell line:primary cells
Time span:48 hours
Number of time points:23

Overview

Macrophages are effector cells of the innate immune system. Their effector function is modulated in response to a wide range of stimuli. Amongst the most studied is lipopolysaccharide (LPS) or endotoxin, the major cell wall component of gram-negative microorganisms. LPS signals through the toll-like receptor 4 (TLR4) to initiate a cascade of transcriptional changes that have been analysed in detail in mice, and which involves waves of gene regulation starting within minutes and extending over 24-48 hours [1,2]. The earliest transcriptional responses, including key pro-inflammatory cytokines, involve the activation of transcriptional elongation from poised RNApolI complexes [3]. Recent studies in the mouse have identified many of the inducible enhancers activated in response to LPS, and the production of eRNAs from these elements [4,5]. However, there are many differences in the transcriptional responses of human and mouse macrophages to LPS [6], reflecting the impact of pathogen selection, and there is also significant variation between individuals. Accordingly, we have studied the response of macrophages from three different individuals.

References:
[1] PMID: 16688168
[2] PMID: 16698233
[3] PMID: 19859064
[4] PMID: 20206554
[5] PMID: 23332752
[6] PMID: 22451944

Sample description

Human monocytes were obtained from anonymised donors with approval of the Human Ethics Committee of the University of Edinburgh (8/9/09). 320mls of blood was extracted from healthy human volunteers and the mononuclear cell fraction was purified using Ficoll gradient centrifugation. The CD14-positive monocytes were purified from the mononuclear cell fraction using magnetic beads (Miltenyi Biotech), and cultured on bacteriological plastic plates in medium (RPMI-1640 plus 10% foetal calf serum) in recombinant human CSF1 (a gift from Chiron) at 100ng/ml. After 7 days the monocyte-derived macrophages were stimulated with 100ng/ml of salmonella R595 LPS. Samples were taken at 0 time, then 15, 30, 45, 60, 80, 100, 120, 150, 180, 210, 240 mins; then 5, 6, 7, 8, 10, 12, 14, 16, 18, 20, 22, 24, 36, 48 hours. Each time course was carried out on one preparation of cells from a single donor, and the precise temporal profile of gene expression varies somewhat between the three replicates. The system is comparable to more limited previous studies based upon microarrays and low-coverage CAGE analysis [6].

References:
[6] PMID: 22451944

Quality control

In each case, the cells respond morphologically to LPS with increased spreading and vacuolation. The response to LPS can be detected through the early induction of classical inflammatory markers, TNF, IL6 and IL1B and late response genes such as IDO1 and CYP27B1 which are induced in human, but not in mouse macrophages [6]. The early response gene IFNB1, which acts in an autocrine manner to induce downstream target genes, is known to vary in its expression between individuals [7]. It was induced somewhat later in Donor 1 than in Donors 2 and 3, but the downstream response was evident in induction of known IFN targets such as MX1.

References:
[6] PMID: 22451944
[7] PMID: 24604202

Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples




Sample Name Time point Donor/Replica
12698-135D7 Monocyte-derived macrophages response to LPS 00hr00min donor1 (t1 Subject1)
12706-135E6 Monocyte-derived macrophages response to LPS 02hr30min donor1 (t9 Subject1)
12707-135E7 Monocyte-derived macrophages response to LPS 03hr00min donor1 (t10 Subject1)
12710-135F1 Monocyte-derived macrophages response to LPS 05hr donor1 (t13 Subject1)
12713-135F4 Monocyte-derived macrophages response to LPS 08hr donor1 (t16 Subject1)
12715-135F6 Monocyte-derived macrophages response to LPS 12hr donor1 (t18 Subject1)
12716-135F7 Monocyte-derived macrophages response to LPS 14hr donor1 (t19 Subject1)
12718-135F9 Monocyte-derived macrophages response to LPS 18hr donor1 (t21 Subject1)
12719-135G1 Monocyte-derived macrophages response to LPS 20hr donor1 (t22 Subject1)
12720-135G2 Monocyte-derived macrophages response to LPS 22hr donor1 (t23 Subject1)
12721-135G3 Monocyte-derived macrophages response to LPS 24hr donor1 (t24 Subject1)
12722-135G4 Monocyte-derived macrophages response to LPS 36hr donor1 (t25 Subject1)
12723-135G5 Monocyte-derived macrophages response to LPS 48hr donor1 (t26 Subject1)
12796-136F6 Monocyte-derived macrophages response to LPS 00hr00min donor2 (t1 Subject2)
12797-136F7 Monocyte-derived macrophages response to LPS 00hr15min donor2 (t2 Subject2)
12798-136F8 Monocyte-derived macrophages response to LPS 00hr30min donor2 (t3 Subject2)
12799-136F9 Monocyte-derived macrophages response to LPS 00hr45min donor2 (t4 Subject2)
12800-136G1 Monocyte-derived macrophages response to LPS 01hr00min donor2 (t5 Subject2)
12801-136G2 Monocyte-derived macrophages response to LPS 01hr20min donor2 (t6 Subject2)
12803-136G4 Monocyte-derived macrophages response to LPS 02hr00min donor2 (t8 Subject2)
12804-136G5 Monocyte-derived macrophages response to LPS 02hr30min donor2 (t9 Subject2)
12805-136G6 Monocyte-derived macrophages response to LPS 03hr00min donor2 (t10 Subject2)
12806-136G7 Monocyte-derived macrophages response to LPS 03hr30min donor2 (t11 Subject2)
12807-136G8 Monocyte-derived macrophages response to LPS 04hr donor2 (t12 Subject2)
12808-136G9 Monocyte-derived macrophages response to LPS 05hr donor2 (t13 Subject2)
12811-136H3 Monocyte-derived macrophages response to LPS 08hr donor2 (t16 Subject2)
12812-136H4 Monocyte-derived macrophages response to LPS 10hr donor2 (t17 Subject2)
12813-136H5 Monocyte-derived macrophages response to LPS 12hr donor2 (t18 Subject2)
12814-136H6 Monocyte-derived macrophages response to LPS 14hr donor2 (t19 Subject2)
12815-136H7 Monocyte-derived macrophages response to LPS 16hr donor2 (t20 Subject2)
12816-136H8 Monocyte-derived macrophages response to LPS 18hr donor2 (t21 Subject2)
12817-136H9 Monocyte-derived macrophages response to LPS 20hr donor2 (t22 Subject2)
12818-136I1 Monocyte-derived macrophages response to LPS 22hr donor2 (t23 Subject2)
12819-136I2 Monocyte-derived macrophages response to LPS 24hr donor2 (t24 Subject2)
12820-136I3 Monocyte-derived macrophages response to LPS 36hr donor2 (t25 Subject2)
12821-136I4 Monocyte-derived macrophages response to LPS 48hr donor2 (t26 Subject2)
12894-137H5 Monocyte-derived macrophages response to LPS 00hr00min donor3 (t1 Subject3)
12895-137H6 Monocyte-derived macrophages response to LPS 00hr15min donor3 (t2 Subject3)
12896-137H7 Monocyte-derived macrophages response to LPS 00hr30min donor3 (t3 Subject3)
12897-137H8 Monocyte-derived macrophages response to LPS 00hr45min donor3 (t4 Subject3)
12898-137H9 Monocyte-derived macrophages response to LPS 01hr00min donor3 (t5 Subject3)
12899-137I1 Monocyte-derived macrophages response to LPS 01hr20min donor3 (t6 Subject3)
12901-137I3 Monocyte-derived macrophages response to LPS 02hr00min donor3 (t8 Subject3)
12902-137I4 Monocyte-derived macrophages response to LPS 02hr30min donor3 (t9 Subject3)
12903-137I5 Monocyte-derived macrophages response to LPS 03hr00min donor3 (t10 Subject3)
12904-137I6 Monocyte-derived macrophages response to LPS 03hr30min donor3 (t11 Subject3)
12905-137I7 Monocyte-derived macrophages response to LPS 04hr donor3 (t12 Subject3)
12906-137I8 Monocyte-derived macrophages response to LPS 05hr donor3 (t13 Subject3)
12909-138A2 Monocyte-derived macrophages response to LPS 08hr donor3 (t16 Subject3)
12910-138A3 Monocyte-derived macrophages response to LPS 10hr donor3 (t17 Subject3)
12911-138A4 Monocyte-derived macrophages response to LPS 12hr donor3 (t18 Subject3)
12912-138A5 Monocyte-derived macrophages response to LPS 14hr donor3 (t19 Subject3)
12913-138A6 Monocyte-derived macrophages response to LPS 16hr donor3 (t20 Subject3)
12914-138A7 Monocyte-derived macrophages response to LPS 18hr donor3 (t21 Subject3)
12915-138A8 Monocyte-derived macrophages response to LPS 20hr donor3 (t22 Subject3)
12916-138A9 Monocyte-derived macrophages response to LPS 22hr donor3 (t23 Subject3)
12917-138B1 Monocyte-derived macrophages response to LPS 24hr donor3 (t24 Subject3)
12918-138B2 Monocyte-derived macrophages response to LPS 36hr donor3 (t25 Subject3)