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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



12698-135D7Monocyte-derived macrophages response to LPS00hr00mindonor1 (t1 Subject1)
12706-135E6Monocyte-derived macrophages response to LPS02hr30mindonor1 (t9 Subject1)
12707-135E7Monocyte-derived macrophages response to LPS03hr00mindonor1 (t10 Subject1)
12710-135F1Monocyte-derived macrophages response to LPS05hrdonor1 (t13 Subject1)
12713-135F4Monocyte-derived macrophages response to LPS08hrdonor1 (t16 Subject1)
12715-135F6Monocyte-derived macrophages response to LPS12hrdonor1 (t18 Subject1)
12716-135F7Monocyte-derived macrophages response to LPS14hrdonor1 (t19 Subject1)
12718-135F9Monocyte-derived macrophages response to LPS18hrdonor1 (t21 Subject1)
12719-135G1Monocyte-derived macrophages response to LPS20hrdonor1 (t22 Subject1)
12720-135G2Monocyte-derived macrophages response to LPS22hrdonor1 (t23 Subject1)
12721-135G3Monocyte-derived macrophages response to LPS24hrdonor1 (t24 Subject1)
12722-135G4Monocyte-derived macrophages response to LPS36hrdonor1 (t25 Subject1)
12723-135G5Monocyte-derived macrophages response to LPS48hrdonor1 (t26 Subject1)
12796-136F6Monocyte-derived macrophages response to LPS00hr00mindonor2 (t1 Subject2)
12797-136F7Monocyte-derived macrophages response to LPS00hr15mindonor2 (t2 Subject2)
12798-136F8Monocyte-derived macrophages response to LPS00hr30mindonor2 (t3 Subject2)
12799-136F9Monocyte-derived macrophages response to LPS00hr45mindonor2 (t4 Subject2)
12800-136G1Monocyte-derived macrophages response to LPS01hr00mindonor2 (t5 Subject2)
12801-136G2Monocyte-derived macrophages response to LPS01hr20mindonor2 (t6 Subject2)
12803-136G4Monocyte-derived macrophages response to LPS02hr00mindonor2 (t8 Subject2)
12804-136G5Monocyte-derived macrophages response to LPS02hr30mindonor2 (t9 Subject2)
12805-136G6Monocyte-derived macrophages response to LPS03hr00mindonor2 (t10 Subject2)
12806-136G7Monocyte-derived macrophages response to LPS03hr30mindonor2 (t11 Subject2)
12807-136G8Monocyte-derived macrophages response to LPS04hrdonor2 (t12 Subject2)
12808-136G9Monocyte-derived macrophages response to LPS05hrdonor2 (t13 Subject2)
12811-136H3Monocyte-derived macrophages response to LPS08hrdonor2 (t16 Subject2)
12812-136H4Monocyte-derived macrophages response to LPS10hrdonor2 (t17 Subject2)
12813-136H5Monocyte-derived macrophages response to LPS12hrdonor2 (t18 Subject2)
12814-136H6Monocyte-derived macrophages response to LPS14hrdonor2 (t19 Subject2)
12815-136H7Monocyte-derived macrophages response to LPS16hrdonor2 (t20 Subject2)
12816-136H8Monocyte-derived macrophages response to LPS18hrdonor2 (t21 Subject2)
12817-136H9Monocyte-derived macrophages response to LPS20hrdonor2 (t22 Subject2)
12818-136I1Monocyte-derived macrophages response to LPS22hrdonor2 (t23 Subject2)
12819-136I2Monocyte-derived macrophages response to LPS24hrdonor2 (t24 Subject2)
12820-136I3Monocyte-derived macrophages response to LPS36hrdonor2 (t25 Subject2)
12821-136I4Monocyte-derived macrophages response to LPS48hrdonor2 (t26 Subject2)
12894-137H5Monocyte-derived macrophages response to LPS00hr00mindonor3 (t1 Subject3)
12895-137H6Monocyte-derived macrophages response to LPS00hr15mindonor3 (t2 Subject3)
12896-137H7Monocyte-derived macrophages response to LPS00hr30mindonor3 (t3 Subject3)
12897-137H8Monocyte-derived macrophages response to LPS00hr45mindonor3 (t4 Subject3)
12898-137H9Monocyte-derived macrophages response to LPS01hr00mindonor3 (t5 Subject3)
12899-137I1Monocyte-derived macrophages response to LPS01hr20mindonor3 (t6 Subject3)
12901-137I3Monocyte-derived macrophages response to LPS02hr00mindonor3 (t8 Subject3)
12902-137I4Monocyte-derived macrophages response to LPS02hr30mindonor3 (t9 Subject3)
12903-137I5Monocyte-derived macrophages response to LPS03hr00mindonor3 (t10 Subject3)
12904-137I6Monocyte-derived macrophages response to LPS03hr30mindonor3 (t11 Subject3)
12905-137I7Monocyte-derived macrophages response to LPS04hrdonor3 (t12 Subject3)
12906-137I8Monocyte-derived macrophages response to LPS05hrdonor3 (t13 Subject3)
12909-138A2Monocyte-derived macrophages response to LPS08hrdonor3 (t16 Subject3)
12910-138A3Monocyte-derived macrophages response to LPS10hrdonor3 (t17 Subject3)
12911-138A4Monocyte-derived macrophages response to LPS12hrdonor3 (t18 Subject3)
12912-138A5Monocyte-derived macrophages response to LPS14hrdonor3 (t19 Subject3)
12913-138A6Monocyte-derived macrophages response to LPS16hrdonor3 (t20 Subject3)
12914-138A7Monocyte-derived macrophages response to LPS18hrdonor3 (t21 Subject3)
12915-138A8Monocyte-derived macrophages response to LPS20hrdonor3 (t22 Subject3)
12916-138A9Monocyte-derived macrophages response to LPS22hrdonor3 (t23 Subject3)
12917-138B1Monocyte-derived macrophages response to LPS24hrdonor3 (t24 Subject3)
12918-138B2Monocyte-derived macrophages response to LPS36hrdonor3 (t25 Subject3)