Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways
Anaplasma phagocytophilum subverts neutrophil function permitting intracellular survival, propagation and transmission. Sustained pro-inflammatory response, recruitment of new host cells for population expansion, and delayed apoptosis are associated with prolonged nuclear presence of NF-κB. We inves...
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doaj-becf03142e394147b4bc1e308cf086212020-11-25T04:00:58ZengFrontiers Media S.A.Frontiers in Public Health2296-25652020-10-01810.3389/fpubh.2020.558283558283Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant PathwaysJ. Stephen Dumler0J. Stephen Dumler1Marguerite Lichay2Wan-Hsin Chen3Kristen E. Rennoll-Bankert4Jin-ho Park5Department of Pathology, F. Edward Hébert School of Medicine, Uniformed Services University for the Health Sciences, Bethesda, MD, United StatesDivision of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United StatesDivision of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United StatesDivision of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United StatesDivision of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United StatesDivision of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United StatesAnaplasma phagocytophilum subverts neutrophil function permitting intracellular survival, propagation and transmission. Sustained pro-inflammatory response, recruitment of new host cells for population expansion, and delayed apoptosis are associated with prolonged nuclear presence of NF-κB. We investigated NF-κB signaling and transcriptional activity with A. phagocytophilum infection using inhibitors of NF-κB signaling pathways, and through silencing of signaling pathway genes. How inhibitors or silencing affected A. phagocytophilum growth, inflammatory response (transcription of the κB-enhanced genes CXCL8 and MMP9), and NF-κB signaling pathway gene expression were tested. Among A. phagocytophilum-infected HL-60 cells, nuclear NF-κB p50, p65, and p52 were detected by immunoblots or iTRAQ proteomics. A. phagocytophilum growth was affected most by the IKKαβ inhibitor wedelolactone (reductions of 96 to 99%) as compared with SC-514 that selectively inhibits IKKβ, illustrating a role for the non-canonical pathway. Wedelolactone inhibited transcription of both CXCL8 (p = 0.001) and MMP9 (p = 0.002) in infected cells. Compared to uninfected THP-1 cells, A. phagocytophilum infection led to >2-fold down regulation of 64 of 92 NF-κB signaling pathway genes, and >2-fold increased expression in only 4. Wedelolactone and SC-514 reversed downregulation in all 64 and 45, respectively, of the genes down-regulated by infection, but decreased expression in 1 gene with SC-514 only. Silencing of 20 NF-κB signal pathway genes increased bacterial growth in 12 (IRAK1, MAP3K1, NFKB1B, MAP3K7, TICAM2, TLR3, TRADD, TRAF3, CHUK, IRAK2, LTBR, and MALT1). Most findings support canonical pathway activation; however, the presence of NFKB2 in infected cell nuclei, selective non-canonical pathway inhibitors that dampen CXCL8 and MMP9 transcription with infection, upregulation of non-canonical pathway target genes CCL13 and CCL19, enhanced bacterial growth with TRAF3 and LTBR silencing provide evidence for non-canonical pathway signaling. Whether this impacts distinct inflammatory processes that underlie disease, and whether and how A. phagocytophilum subverts NF-κB signaling via these pathways, need to be investigated.https://www.frontiersin.org/articles/10.3389/fpubh.2020.558283/fullAnaplasma phagocytophilumNF-κBsignalingneutrophilinflammation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
J. Stephen Dumler J. Stephen Dumler Marguerite Lichay Wan-Hsin Chen Kristen E. Rennoll-Bankert Jin-ho Park |
spellingShingle |
J. Stephen Dumler J. Stephen Dumler Marguerite Lichay Wan-Hsin Chen Kristen E. Rennoll-Bankert Jin-ho Park Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways Frontiers in Public Health Anaplasma phagocytophilum NF-κB signaling neutrophil inflammation |
author_facet |
J. Stephen Dumler J. Stephen Dumler Marguerite Lichay Wan-Hsin Chen Kristen E. Rennoll-Bankert Jin-ho Park |
author_sort |
J. Stephen Dumler |
title |
Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways |
title_short |
Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways |
title_full |
Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways |
title_fullStr |
Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways |
title_full_unstemmed |
Anaplasma phagocytophilum Activates NF-κB Signaling via Redundant Pathways |
title_sort |
anaplasma phagocytophilum activates nf-κb signaling via redundant pathways |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Public Health |
issn |
2296-2565 |
publishDate |
2020-10-01 |
description |
Anaplasma phagocytophilum subverts neutrophil function permitting intracellular survival, propagation and transmission. Sustained pro-inflammatory response, recruitment of new host cells for population expansion, and delayed apoptosis are associated with prolonged nuclear presence of NF-κB. We investigated NF-κB signaling and transcriptional activity with A. phagocytophilum infection using inhibitors of NF-κB signaling pathways, and through silencing of signaling pathway genes. How inhibitors or silencing affected A. phagocytophilum growth, inflammatory response (transcription of the κB-enhanced genes CXCL8 and MMP9), and NF-κB signaling pathway gene expression were tested. Among A. phagocytophilum-infected HL-60 cells, nuclear NF-κB p50, p65, and p52 were detected by immunoblots or iTRAQ proteomics. A. phagocytophilum growth was affected most by the IKKαβ inhibitor wedelolactone (reductions of 96 to 99%) as compared with SC-514 that selectively inhibits IKKβ, illustrating a role for the non-canonical pathway. Wedelolactone inhibited transcription of both CXCL8 (p = 0.001) and MMP9 (p = 0.002) in infected cells. Compared to uninfected THP-1 cells, A. phagocytophilum infection led to >2-fold down regulation of 64 of 92 NF-κB signaling pathway genes, and >2-fold increased expression in only 4. Wedelolactone and SC-514 reversed downregulation in all 64 and 45, respectively, of the genes down-regulated by infection, but decreased expression in 1 gene with SC-514 only. Silencing of 20 NF-κB signal pathway genes increased bacterial growth in 12 (IRAK1, MAP3K1, NFKB1B, MAP3K7, TICAM2, TLR3, TRADD, TRAF3, CHUK, IRAK2, LTBR, and MALT1). Most findings support canonical pathway activation; however, the presence of NFKB2 in infected cell nuclei, selective non-canonical pathway inhibitors that dampen CXCL8 and MMP9 transcription with infection, upregulation of non-canonical pathway target genes CCL13 and CCL19, enhanced bacterial growth with TRAF3 and LTBR silencing provide evidence for non-canonical pathway signaling. Whether this impacts distinct inflammatory processes that underlie disease, and whether and how A. phagocytophilum subverts NF-κB signaling via these pathways, need to be investigated. |
topic |
Anaplasma phagocytophilum NF-κB signaling neutrophil inflammation |
url |
https://www.frontiersin.org/articles/10.3389/fpubh.2020.558283/full |
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