Limits...
RSV-Induced H3K4 Demethylase KDM5B Leads to Regulation of Dendritic Cell-Derived Innate Cytokines and Exacerbates Pathogenesis In Vivo.

Ptaschinski C, Mukherjee S, Moore ML, Albert M, Helin K, Kunkel SL, Lukacs NW - PLoS Pathog. (2015)

Bottom Line: In vivo, infection of Kdm5bfl/fl-CD11c-Cre+ mice with RSV resulted in higher production of IFN-γ and reduced IL-4 and IL-5 compared to littermate controls, with significantly decreased inflammation, IL-13, and mucus production in the lungs.Importantly, human monocyte-derived DCs treated with a chemical inhibitor for KDM5B resulted in increased innate cytokine levels as well as elicited decreased Th2 cytokines when co-cultured with RSV reactivated CD4+ T cells.These results suggest that KDM5B acts to repress type I IFN and other innate cytokines to promote an altered immune response following RSV infection that contributes to development of chronic disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Michigan, Ann Arbor, Michigan, United States of America.

ABSTRACT
Respiratory syncytial virus (RSV) infection can result in severe disease partially due to its ability to interfere with the initiation of Th1 responses targeting the production of type I interferons (IFN) and promoting a Th2 immune environment. Epigenetic modulation of gene transcription has been shown to be important in regulating inflammatory pathways. RSV-infected bone marrow-derived DCs (BMDCs) upregulated expression of Kdm5b/Jarid1b H3K4 demethylase. Kdm5b-specific siRNA inhibition in BMDC led to a 10-fold increase in IFN-β as well as increases in IL-6 and TNF-α compared to control-transfected cells. The generation of Kdm5bfl/fl-CD11c-Cre+ mice recapitulated the latter results during in vitro DC activation showing innate cytokine modulation. In vivo, infection of Kdm5bfl/fl-CD11c-Cre+ mice with RSV resulted in higher production of IFN-γ and reduced IL-4 and IL-5 compared to littermate controls, with significantly decreased inflammation, IL-13, and mucus production in the lungs. Sensitization with RSV-infected DCs into the airways of naïve mice led to an exacerbated response when mice were challenged with live RSV infection. When Kdm5b was blocked in DCs with siRNA or DCs from Kdm5bfl/fl-CD11c-CRE mice were used, the exacerbated response was abrogated. Importantly, human monocyte-derived DCs treated with a chemical inhibitor for KDM5B resulted in increased innate cytokine levels as well as elicited decreased Th2 cytokines when co-cultured with RSV reactivated CD4+ T cells. These results suggest that KDM5B acts to repress type I IFN and other innate cytokines to promote an altered immune response following RSV infection that contributes to development of chronic disease.

No MeSH data available.


Related in: MedlinePlus

Kdm5b expression increases following infection of BMDCs with RSV.Bone marrow-derived dendritic cells were infected with RSV (MOI = 1) or treated with poly(I:C) (20 μg/ml) or imiquimod (1 μg/ml) for four hours. RNA was extracted and reverse transcribed, and the cDNA was used in a PCR array to measure changes in the expression of epigenetic enzymes (A), including the histone lysine demethylase family. Expression of Kdm5b was measured over a time course following infection with RSV (B). n = 3–5 samples/group, and data are representative of three independent experiments. *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4470918&req=5

ppat.1004978.g001: Kdm5b expression increases following infection of BMDCs with RSV.Bone marrow-derived dendritic cells were infected with RSV (MOI = 1) or treated with poly(I:C) (20 μg/ml) or imiquimod (1 μg/ml) for four hours. RNA was extracted and reverse transcribed, and the cDNA was used in a PCR array to measure changes in the expression of epigenetic enzymes (A), including the histone lysine demethylase family. Expression of Kdm5b was measured over a time course following infection with RSV (B). n = 3–5 samples/group, and data are representative of three independent experiments. *p<0.05.

Mentions: A previous report has identified a role for epigenetic regulation in immune cells following viral infection [21]. As DCs are critical for priming the T cell response to RSV infection, studies were initiated to determine whether exposing DCs to RSV resulted in changes in the expression of epigenetic factors in the DCs. BMDCs were infected with RSV or activated by p(I:C) or imiquimod, the ligands for TLR3 and TLR7 respectively, as RSV is known to activate cells through both TLR3 and TLR7 [22,23], in addition to other mechanisms. In order to observe early gene expression of epigenetic enzymes, RNA was harvested at 4 hours post treatment to examine transcription levels of genes coding for “epigenetic” enzymes by qPCR array. Several classes of enzymes were analyzed including histone deacetylases (HDACs), histone lysine demethylases (KDMs), protein arginine methyltransferases (PRMTs), and histone lysine methyltransferases (KMTs) (Fig 1A). A defining observation was the upregulation of Kdm5b demethylase by RSV in contrast to the downregulation of this enzyme by stimulation through TLR3 and TLR7 (Fig 1A). While Kdm6b was upregulated by RSV infection of DCs, this enzyme was also significantly upregulated by treatment of cells with imiquimod. Because Kdm5b was upregulated only by RSV, studies focused on Kdm5b as a potential unique enzyme in the DC response to RSV. PCR analysis confirmed the peak expression of Kdm5b in BMDCs at 12 hours following RSV infection (Fig 1B). Furthermore, while Kdm5b was upregulated in BMDCs infected with RSV, it was not upregulated by influenza (H1N1) virus, nor in RSV-infected epithelial cells or alveolar macrophages (S1 Fig). Therefore, studies focused on H3K4 demethylase Kdm5b and its role on perturbing critical innate immune genes in DCs.


RSV-Induced H3K4 Demethylase KDM5B Leads to Regulation of Dendritic Cell-Derived Innate Cytokines and Exacerbates Pathogenesis In Vivo.

Ptaschinski C, Mukherjee S, Moore ML, Albert M, Helin K, Kunkel SL, Lukacs NW - PLoS Pathog. (2015)

Kdm5b expression increases following infection of BMDCs with RSV.Bone marrow-derived dendritic cells were infected with RSV (MOI = 1) or treated with poly(I:C) (20 μg/ml) or imiquimod (1 μg/ml) for four hours. RNA was extracted and reverse transcribed, and the cDNA was used in a PCR array to measure changes in the expression of epigenetic enzymes (A), including the histone lysine demethylase family. Expression of Kdm5b was measured over a time course following infection with RSV (B). n = 3–5 samples/group, and data are representative of three independent experiments. *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470918&req=5

ppat.1004978.g001: Kdm5b expression increases following infection of BMDCs with RSV.Bone marrow-derived dendritic cells were infected with RSV (MOI = 1) or treated with poly(I:C) (20 μg/ml) or imiquimod (1 μg/ml) for four hours. RNA was extracted and reverse transcribed, and the cDNA was used in a PCR array to measure changes in the expression of epigenetic enzymes (A), including the histone lysine demethylase family. Expression of Kdm5b was measured over a time course following infection with RSV (B). n = 3–5 samples/group, and data are representative of three independent experiments. *p<0.05.
Mentions: A previous report has identified a role for epigenetic regulation in immune cells following viral infection [21]. As DCs are critical for priming the T cell response to RSV infection, studies were initiated to determine whether exposing DCs to RSV resulted in changes in the expression of epigenetic factors in the DCs. BMDCs were infected with RSV or activated by p(I:C) or imiquimod, the ligands for TLR3 and TLR7 respectively, as RSV is known to activate cells through both TLR3 and TLR7 [22,23], in addition to other mechanisms. In order to observe early gene expression of epigenetic enzymes, RNA was harvested at 4 hours post treatment to examine transcription levels of genes coding for “epigenetic” enzymes by qPCR array. Several classes of enzymes were analyzed including histone deacetylases (HDACs), histone lysine demethylases (KDMs), protein arginine methyltransferases (PRMTs), and histone lysine methyltransferases (KMTs) (Fig 1A). A defining observation was the upregulation of Kdm5b demethylase by RSV in contrast to the downregulation of this enzyme by stimulation through TLR3 and TLR7 (Fig 1A). While Kdm6b was upregulated by RSV infection of DCs, this enzyme was also significantly upregulated by treatment of cells with imiquimod. Because Kdm5b was upregulated only by RSV, studies focused on Kdm5b as a potential unique enzyme in the DC response to RSV. PCR analysis confirmed the peak expression of Kdm5b in BMDCs at 12 hours following RSV infection (Fig 1B). Furthermore, while Kdm5b was upregulated in BMDCs infected with RSV, it was not upregulated by influenza (H1N1) virus, nor in RSV-infected epithelial cells or alveolar macrophages (S1 Fig). Therefore, studies focused on H3K4 demethylase Kdm5b and its role on perturbing critical innate immune genes in DCs.

Bottom Line: In vivo, infection of Kdm5bfl/fl-CD11c-Cre+ mice with RSV resulted in higher production of IFN-γ and reduced IL-4 and IL-5 compared to littermate controls, with significantly decreased inflammation, IL-13, and mucus production in the lungs.Importantly, human monocyte-derived DCs treated with a chemical inhibitor for KDM5B resulted in increased innate cytokine levels as well as elicited decreased Th2 cytokines when co-cultured with RSV reactivated CD4+ T cells.These results suggest that KDM5B acts to repress type I IFN and other innate cytokines to promote an altered immune response following RSV infection that contributes to development of chronic disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Michigan, Ann Arbor, Michigan, United States of America.

ABSTRACT
Respiratory syncytial virus (RSV) infection can result in severe disease partially due to its ability to interfere with the initiation of Th1 responses targeting the production of type I interferons (IFN) and promoting a Th2 immune environment. Epigenetic modulation of gene transcription has been shown to be important in regulating inflammatory pathways. RSV-infected bone marrow-derived DCs (BMDCs) upregulated expression of Kdm5b/Jarid1b H3K4 demethylase. Kdm5b-specific siRNA inhibition in BMDC led to a 10-fold increase in IFN-β as well as increases in IL-6 and TNF-α compared to control-transfected cells. The generation of Kdm5bfl/fl-CD11c-Cre+ mice recapitulated the latter results during in vitro DC activation showing innate cytokine modulation. In vivo, infection of Kdm5bfl/fl-CD11c-Cre+ mice with RSV resulted in higher production of IFN-γ and reduced IL-4 and IL-5 compared to littermate controls, with significantly decreased inflammation, IL-13, and mucus production in the lungs. Sensitization with RSV-infected DCs into the airways of naïve mice led to an exacerbated response when mice were challenged with live RSV infection. When Kdm5b was blocked in DCs with siRNA or DCs from Kdm5bfl/fl-CD11c-CRE mice were used, the exacerbated response was abrogated. Importantly, human monocyte-derived DCs treated with a chemical inhibitor for KDM5B resulted in increased innate cytokine levels as well as elicited decreased Th2 cytokines when co-cultured with RSV reactivated CD4+ T cells. These results suggest that KDM5B acts to repress type I IFN and other innate cytokines to promote an altered immune response following RSV infection that contributes to development of chronic disease.

No MeSH data available.


Related in: MedlinePlus