Dataset Information

IL-4-primed bone marrow macrophages isolated from C57Bl6/J or Balb/cAnNCrl mice

ABSTRACT: To compare the expression profile of differentiated mouse bone marrow macrophages (BMM) in response to IL-4, we have employed whole genome microarray expression profiling. For this purpose, bone marrow cells were isolated from 8 to 12 weeks old C57BL6/J and Balb/cAnNCrl mice and cultured in the presence of the macrophage colony-stimulating factor (M-CSF). After seven days of culture, IL-4 was added for 4 and 18 hours. Keywords: Mice strain comparison; Gene expression profiling

ORGANISM(S): Mus musculus

PROVIDER: GSE14644 | GEO | 2009/08/10

SECONDARY ACCESSION(S): PRJNA112305

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:ABSTRACT Macrophages are essential components of the innate immune system and crucial for pathogen elimination in early stages of infection. We previously observed that bone marrow-derived macrophages (BMM) from C57BL/6 mice exhibited increased killing activity against Burkholderia pseudomallei compared to BMM from BALB/c mice. This effect was particularly pronounced when cells were treated with IFN-M-NM-3. To unravel mechanisms that could explain these distinct bactericidal effects, a comparative combined proteome and transcriptome analysis of untreated and IFN-M-NM-3 treated BALB/c and C57BL/6 BMM under standardized serum-free conditions was carried out. We found differences in gene expression/ protein abundance belonging to cellular oxidative and antioxidative stress systems. Genes/ proteins involved in the generation of oxidant molecules and the function of phagosomes (respiratory chain ATPase, lysosomal enzymes, cathepsin) were predominantly higher expressed/ more abundant in C57BL/6 BMM. Components involved in alleviation of oxidative stress (peroxiredoxin, mitochondrial superoxide dismutase) were more abundant in C57BL/6 BMM as well. Thus, C57BL/6 BMM seemed to be better equipped with cellular systems that may be advantageous in combating engulfed pathogens. Simultaneously, C57BL/6 BMM were well protected from oxidative burst. We assume that these variations co-determine differences in resistance between BALB/c and C57BL/6 mice observed in many infection models. Biological significance: In this study we performed combined transcriptome and proteome analyses on BMM derived from two inbred mouse strains that are frequently used for studies in the field of host-pathogen interaction research. Strain differences between BALB/c and C57BL/6 BMM were found to originate mainly from different protein abundance levels rather than from different gene expression. Differences in abundance of respiratory chain complexes and phagosomal proteins as well as differential regulation of components belonging to various antioxidant stress systems help to explain long-known differences between the mouse strains concerning their different susceptibility in several infection models. In this study 12 samples were analyzed. They are classified into four groups: BMM of Balb/c mice medium control, BMM of Balb/c mice interferon gamma treated, BMM of C57Bl/6 mice medium control, and BMM of C57Bl/6 mice interferon gamma treated. Each of the four groups contains 3 biological replicates.

Project description:ABSTRACT Macrophages are essential components of the innate immune system and crucial for pathogen elimination in early stages of infection. We previously observed that bone marrow-derived macrophages (BMM) from C57BL/6 mice exhibited increased killing activity against Burkholderia pseudomallei compared to BMM from BALB/c mice. This effect was particularly pronounced when cells were treated with IFN-γ. To unravel mechanisms that could explain these distinct bactericidal effects, a comparative combined proteome and transcriptome analysis of untreated and IFN-γ treated BALB/c and C57BL/6 BMM under standardized serum-free conditions was carried out. We found differences in gene expression/ protein abundance belonging to cellular oxidative and antioxidative stress systems. Genes/ proteins involved in the generation of oxidant molecules and the function of phagosomes (respiratory chain ATPase, lysosomal enzymes, cathepsin) were predominantly higher expressed/ more abundant in C57BL/6 BMM. Components involved in alleviation of oxidative stress (peroxiredoxin, mitochondrial superoxide dismutase) were more abundant in C57BL/6 BMM as well. Thus, C57BL/6 BMM seemed to be better equipped with cellular systems that may be advantageous in combating engulfed pathogens. Simultaneously, C57BL/6 BMM were well protected from oxidative burst. We assume that these variations co-determine differences in resistance between BALB/c and C57BL/6 mice observed in many infection models. Biological significance: In this study we performed combined transcriptome and proteome analyses on BMM derived from two inbred mouse strains that are frequently used for studies in the field of host-pathogen interaction research. Strain differences between BALB/c and C57BL/6 BMM were found to originate mainly from different protein abundance levels rather than from different gene expression. Differences in abundance of respiratory chain complexes and phagosomal proteins as well as differential regulation of components belonging to various antioxidant stress systems help to explain long-known differences between the mouse strains concerning their different susceptibility in several infection models.

Project description:Interferon (IFN)γ and interleukin (IL)-4 are central regulators of T helper 1 (Th1) and T helper 2 (Th2) immune responses, respectively. Both cytokines have a major impact on macrophage phenotypes: IFNγ–priming and subsequent TLR4 activation induces so called classically activated macrophages that are characterized by pronounced pro-inflammatory responses, whereas IL-4–treated macrophages, commonly called alternatively activated, are known to develop enhanced capacity for endocytosis, antigen presentation, and tissue repair and are generally considered anti-inflammatory. Considering IL-4 as priming rather than activating stimulus, we now compared the TLR4–dependent global gene activation program in IFNγ– versus IL-4–pretreated mouse macrophages, which has rarely been studied so far. Although both cytokines frequently induced opposing effects on gene transcription, the subsequent activation of bone marrow-derived macrophages by lipopolysaccharide (LPS) produced a strong, priming dependent pro-inflammatory response in both macrophage types. For example, the production of key pro-inflammatory cytokines IL-6 and IL-12 was significantly higher in IL-4– versus IFNγ–primed macrophages and several cytokine genes, including Il19, Ccl17, Ccl22, Ccl24 and Cxcl5, were preferentially induced in alternatively primed and LPS activated mouse macrophages. In a subset of genes, including IL12a, IFNγ priming was actually found to suppress LPS–induced gene expression in a Stat1–dependent manner. Our data suggest that IL-4–priming is not per se anti-inflammatory but generates a macrophage that is “tissue protective” but still capable of mounting a strong inflammatory response after TLR4–dependent activation. Keywords: Gene expression profiling Gene expression was investigated in mouse bone marrow-derived macrophages (BMM). On day 7, BMM were stimulated with either IL-4 or IFNγ overnight (18h in total). LPS treatment was performed in primed and unprimed macrophages 4 h prior to harvesting. At least three independent experiments were performed for each condition.

Dataset Information

IL-4-primed bone marrow macrophages isolated from C57Bl6/J or Balb/cAnNCrl mice

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets