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Bone Marrow-derived Macrophage Production

Bone Marrow-derived Macrophage Production

Published on: November 22, 2013

Transcriptomic identification of genes associated with thrombosis and coagulation in lipopolysaccharide-exposed

Hannah R Cohen¹, Michelle A DeCourcey², William C Davis³

¹Department of Veterinary Clinical Sciences, Washington State University, Pullman, WA.

American Journal of Veterinary Research

|September 30, 2024

Insights

Bovine monocyte-derived macrophages (MoMΦ) exposed to lipopolysaccharide (LPS) show complex gene expression changes, involving both procoagulant and anticoagulant pathways. This study reveals MoMΦ

Area of Science:

Immunology
Molecular Biology
Genomics

Background:

Sepsis can lead to coagulopathies, affecting blood clotting.
Monocyte-derived macrophages (MoMΦ) play a role in the immune response to pathogens like LPS.
Understanding early cellular responses is crucial for sepsis management.

Purpose of the Study:

To investigate gene expression changes in bovine MoMΦ stimulated with LPS.
To analyze differential expression of genes related to coagulation pathways.
To test the hypothesis of upregulated procoagulant and downregulated anticoagulant genes.

Main Methods:

Isolation of MoMΦ from Holstein steers.
In vitro exposure of MoMΦ to lipopolysaccharide (LPS) or control for 3 hours.
Transcriptomic analysis using RNA sequencing to identify differentially expressed genes.

Main Results:

1,602 genes upregulated and 1,209 downregulated post-LPS exposure.
Significant upregulation of proinflammatory, anti-inflammatory, procoagulant, and anticoagulant genes.
Downregulation of one gene involved in coagulation promotion.

Conclusions:

MoMΦ exhibit a multifaceted response to LPS, balancing procoagulant and anticoagulant gene expression.
This complex response highlights the interplay of pathways in early sepsis.
Bovine MoMΦ serve as a valuable model for studying sepsis-associated coagulopathies.

Read full analysis

Abstract

Objective:

The objective of this study was to investigate the differential expression of genes associated with coagulation in bovine monocyte-derived macrophages (MoMΦ) exposed to lipopolysaccharide (LPS) in vitro. We hypothesized that MoMΦ stimulated with LPS would have upregulation of procoagulant genes and downregulation of genes protecting against coagulation.

Methods:

MoMΦ were isolated from Holstein steers and exposed to Escherichia coli-derived LPS or a control for 3 hours. We used transcriptomics (RNA sequencing) to characterize the differential expression of genes associated with coagulation in the LPS-exposed MoMΦ relative to the control group.

Results:

1,602 genes were upregulated and 1,209 genes downregulated 3 hours after exposure to LPS. Monocyte-derived macrophages exposed to LPS displayed statistically significant upregulation of 4 proinflammatory genes, 2 anti-inflammatory genes, 8 genes involved in promoting coagulation, and 5 genes considered protective against coagulation. There was significant downregulation of 1 gene involved in the promotion of coagulation.

Conclusions:

Our results showed increased expression of most genes investigated promoting and protecting against coagulation and increased expression of proinflammatory and anti-inflammatory genes. These findings suggest that MoMΦ exhibit a multifaceted response in the early response to LPS, promoting and protecting against excessive coagulation. This multifaceted response highlights the interplay between different pathways involved in early sepsis.

Clinical Relevance:

Our data demonstrate the utility of using MoMΦ as a model system to investigate sepsis-associated coagulopathies. These insights into the early transcriptomic changes in response to LPS may help guide future research on the development of treatment modalities or diagnostic tests for patients with sepsis.

Keywords:

bovine coagulation lipopolysaccharide monocyte-derived macrophages sepsis

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