The Atypical Inhibitor of NF-κB, IκBζ, Controls Macrophage Interleukin-10 Expression

Hörber, Sebastian; Hildebrand, Dominic G.; Lieb, Wolfgang; Lorscheid, Sebastian; Hailfinger, Stephan; Schulze‐Osthoff, Klaus; Eßmann, Frank

doi:10.1074/jbc.m116.718825

Cited by 42 publications

(38 citation statements)

References 51 publications

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“…RNA was reverse transcribed using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). For real-time PCR, gene-specific primers for iNOS [ 16 ], Arg I [ 17 ], COX-2 [ 18 ], TNF-α [ 19 ], Fizz1/RELMα , Ym-1 [ 20 ], SPHK1 , LIGHT [ 21 ], MertK [ 22 ], IFN-ɣ , IL-2 , IL-4 [ 23 ], IL-10 [ 24 ], and GATA-3 [ 25 ] were used. To detect IL-6 and 18S RNA expression, the following primers were used: IL-6 forward 5′-CCACTTCACAAGTCGGAGGCTTA-3′; IL-6 reverse 5′-GCAAGTGCATCATCGTTGTTCATAC-3′; 18S RNA forward 5′-CGGCTACCACATCCAAGGAA-3′, 18S RNA reverse 5′-GCTGGAATTACCGCGGCT-3′.…”

Section: Methodsmentioning

confidence: 99%

Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization

et al. 2018

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BackgroundDuring the last decade, mesenchymal stem cells (MSCs) have gained much attention in the field of regenerative medicine due to their capacity to differentiate into different cell types and to promote immunosuppressive effects. However, the underlying mechanism of MSC-mediated immunoregulation is not fully understood so far. Macrophages are distinguished in classical activated, pro-inflammatory M1 and alternatively activated M2 cells, which possess different functions and transcriptional profiles with respect to inflammatory responses. As polarization is not fixed, macrophage functional plasticity might be modulated by the microenvironment allowing them to rapidly react to danger signals and maintaining tissue homeostasis.MethodsMurine MSCs were preconditioned with IL-1ß and IFN-ɣ to enhance their immunosuppressive capacity regarding macrophage polarization under M1- and M2a-polarizing conditions. Macrophage polarization was analyzed by real-time PCR, flow cytometry, and cytokine detection in culture supernatants. The role of MSC-derived nitric oxide (NO), prostaglandin E2 (PGE2), and IL-6 in this process has been evaluated using siRNA transfection and IL-6 receptor-deficient macrophages, respectively.ResultsPreconditioned, but not unprimed, MSCs secreted high levels of NO, IL-6, and PGE2. Co-culture with macrophages (M0) in the presence of M1 inducers (LPS + IFN-ɣ) led to significant reduction of CD86 and iNOS protein in macrophages and diminished TNF-α secretion. Additionally, CD86 and iNOS protein expression as well as NO and IL-10 secretion were markedly increased under M2a-polarizing culture conditions (IL-4). MSC-dependent macrophage polarization did not depend on direct cell-cell contact. Co-culturing in the presence of LPS and IFN-ɣ resulted in the upregulation of M2a, M2b, and M2c marker genes, whereas in the presence of IL-4 only M2b markers were significantly increased. In turn, IL-10-producing regulatory M2b cells significantly inhibited IFN-ɣ expression in CD4+ T lymphocytes. Finally, we show that MSC-mediated macrophage polarization strongly depends on IL-6, whereas a minor role for NO and PGE2 was found.ConclusionsPreconditioning of MSCs highly strengthens their capacity to regulate macrophage features and to promote immunosuppression. Repression of M1 polarization during inflammation and M2b polarization under anti-inflammatory conditions strongly depend on functional IL-6 signaling in macrophages. The potential benefit of preconditioned MSCs and IL-6 should be considered for future clinical treatment.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-1039-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization

et al. 2018

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“…IB 1/16). Organs were excised at the indicated times post-irradiation and bone marrow was isolated as described [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

Simultaneous quantification of DNA damage and mitochondrial copy number by long-run DNA-damage quantification (LORD-Q)

et al. 2017

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DNA damage and changes in the mitochondrial DNA content have been implicated in ageing and cancer development. To prevent genomic instability and tumorigenesis, cells must maintain the integrity of their nuclear and mitochondrial DNA. Advances in the research of DNA damage protection and genomic stability, however, also depend on the availability of techniques that can reliably quantify alterations of mitochondrial DNA copy numbers and DNA lesions in an accurate high-throughput manner. Unfortunately, no such method has been established yet. Here, we describe the high-sensitivity long-run real-time PCR technique for DNA-damage quantification (LORD-Q) and its suitability to simultaneously measure DNA damage rates and mitochondrial DNA copy numbers in cultured cells and tissue samples. Using the LORD-Q multiplex assay, we exemplarily show that the mitochondrial DNA content does not directly affect DNA damage susceptibility, but influences the efficacy of certain anticancer drugs. Hence, LORD-Q provides a fast and precise method to assess DNA lesions, DNA repair and mtDNA replication as well as their role in a variety of pathological settings.

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“…In addition, different T H cell subsets produce IL-10 in response to distinct combinations of cytokines (Gabryšová et al, 2014). These signals lead to the binding of diverse transcription factors at various promoter and enhancer elements within the Il10 locus to activate transcription within myeloid and lymphoid cells (Saraiva and O'Garra, 2010;Gabryšová et al, 2014;Hörber et al, 2016). Much less is known about transcriptional pathways that limit the production of IL-10 (Iyer and Cheng, 2012).…”

Section: Introductionmentioning

confidence: 99%

Bhlhe40 is an essential repressor of IL-10 during Mycobacterium tuberculosis infection

Huynh

Lin

Kimmey

et al. 2018

Journal of Experimental Medicine

102

138

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The cytokine IL-10 antagonizes pathways that control () infection. Nevertheless, the impact of IL-10 during infection has been difficult to decipher because loss-of-function studies in animal models have yielded only mild phenotypes. We have discovered that the transcription factor basic helix-loop-helix family member e40 (Bhlhe40) is required to repress expression during infection. Loss of Bhlhe40 in mice results in higher expression, higher bacterial burden, and early susceptibility similar to that observed in mice lacking IFN-γ. Deletion of in mice reverses these phenotypes. Bhlhe40 deletion in T cells or CD11c cells is sufficient to cause susceptibility to Bhlhe40 represents the first transcription factor found to be essential during infection to specifically regulate expression, revealing the importance of strict control of IL-10 production by innate and adaptive immune cells during infection. Our findings uncover a previously elusive but significant role for IL-10 in pathogenesis.

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The Atypical Inhibitor of NF-κB, IκBζ, Controls Macrophage Interleukin-10 Expression

Cited by 42 publications

References 51 publications

Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization

Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization

Simultaneous quantification of DNA damage and mitochondrial copy number by long-run DNA-damage quantification (LORD-Q)

Bhlhe40 is an essential repressor of IL-10 during Mycobacterium tuberculosis infection

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