Bcl10 mediates LPS-induced activation of NF-κB and IL-8 in human intestinal epithelial cells

Bhattacharyya, Sumit; Borthakur, Alip; Pant, Nitika; Dudeja, Pradeep K.; Tobacman, Joanne K.

doi:10.1152/ajpgi.00149.2007

Cited by 42 publications

(58 citation statements)

References 39 publications

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“…BCL10 silencing inhibited the CGN-induced effects on phospho-NIK. These data challenge some previous observations about NIK activation, because previously, the LPS-activated TLR-mediated pathway was not associated with NIK (29), and we have found that the TLR4-BCL10 mediated pathway resembles the CGNactivated pathway (16). The data demonstrated that BCL10 silencing inhibited the increase in phospho-NIK, consistent with a vital role for BCL10 in the IKK␣-mediated pathway, although the role of phospho-NIK in the IKK␤-mediated pathway remains obscure.…”

Section: Discussioncontrasting

confidence: 78%

“…Control or treated cells were lysed in RIPA buffer (50 mM Tris containing 0.15 M NaCl, 1% Nonidet P-40, 0.5% deoxycholic acid, and 0.1% SDS, pH 7.4), and the cell extracts were stored at Ϫ80°C until assayed, using ELISA as reported previously (16 Determinations of NF-B p65, p50, p52, RelB, c-Rel, and p100-Nuclear extracts were prepared from treated and control NCM460 cells as described previously, using a nuclear extraction kit (Active Motif, Carlsbad, CA). Activated NF-B family members in the samples were determined by oligonucleotidebased ELISA (Active Motif).…”

Section: Ncm460 Cells and Mouse Embryonic Fibroblasts In Tissuementioning

confidence: 99%

“…Silencing of BCL10 mRNA Expression-Silencing BCL10 was performed as described previously (6,16). Effectiveness of BCL10 silencing in the NCM460 cells and the MEF was determined by BCL10 ELISA, following exposure to BCL10 siRNA for 24 h. In the NCM460 cells, silencing reduced baseline BCL10 protein by ϳ82% (from 1.30 (Ϯ0.07) to 0.24 (Ϯ0.02) ng/mg protein).…”

Section: Ncm460 Cells and Mouse Embryonic Fibroblasts In Tissuementioning

confidence: 99%

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B-cell CLL/Lymphoma 10 (BCL10) Is Required for NF-κB Production by Both Canonical and Noncanonical Pathways and for NF-κB-inducing Kinase (NIK) Phosphorylation

Bhattacharyya

Borthakur

Tyagi

et al. 2010

Journal of Biological Chemistry

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The importance of activation of B-cell lymphoma/CLL 10 (BCL10) 2 has been recognized previously in the mucosa-associated lymphoid tissue (MALT) lymphomas, in which translocations involving MALT1 lead to constitutive activation of BCL10 and NF-B transcription (1, 2). Other reports have defined a critical role for BCL10 in the inflammatory cascade in epithelial cells, including activation by lysophosphatidic acid (3), G-protein-coupled receptor (4), and angiotensin II (5). In this report, we demonstrate that BCL10 was required for NF-B activation by both canonical and noncanonical pathways, following stimulation by the sulfated polysaccharide carrageenan (CGN) in mouse embryonic fibroblasts and in human colonic epithelial cells. Previously, we reported that CGN significantly up-regulated transcription of BCL10 in NCM460 cells (6, 7). The sulfated polysaccharide carrageenan has been widely used for decades to induce inflammation in animal and tissue culture models. In our previous work, CGN was shown to induce an inflammatory cascade in human colonic epithelial cells by two distinct mechanisms: an immune-mediated mechanism involving TLR4 (toll-like receptor 4), IRAK (IL-1␤ receptor activating kinase), BCL10, phospho-IB␣ (inhibitor of B), NF-B (nuclear factor B), and IL-8 (interleukin-8); and a reactive oxygen species (ROS)-mediated mechanism involving phospho-Hsp27, IB kinase (IKK)-␤, phospho-IB␣, BCL10, NF-B, and . Carrageenan activation of these pathways was attributable to its distinctive chemical structure, including its resemblance to the naturally occurring glycosaminoglycans, its highly sulfated galactose residues, and its unusual ␣-Gal(133)Gal bond that is a known immune epitope (10, 11). Because CGN is a commonly used food additive in the Western diet, these pathways may induce inflammation and disease in the human colon and implicate a role for BCL10 in human disease, in addition to the MALT lymphomas.To further define the BCL10-mediated activation of NF-B and the interactions between the ROS and TLR4-BCL10 pathways, the requirements for different components of the IKK signaling complex and the responses of different members of the NF-B family were investigated. The IKK signaling complex, including the catalytic components IKK␣ and IKK␤ and the regulatory component IKK␥, also known as NEMO (NF-B essential modifying factor), integrates upstream signals and leads to the phosphorylation of IB␣. Subsequently, phospho-IB␣ is ubiquitinated, and the localization signal for NF-B nuclear translocation is exposed. These events represent critical signals in the progression of the inflammatory cascade from * This work was supported in part by the Department of Veterans Affairs (to J. K. T.) and by NIDDK, National Institutes of Health Grants DK68324 and DK54016 (to P. K. D.

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Section: Discussioncontrasting

confidence: 78%

Section: Ncm460 Cells and Mouse Embryonic Fibroblasts In Tissuementioning

confidence: 99%

Section: Ncm460 Cells and Mouse Embryonic Fibroblasts In Tissuementioning

confidence: 99%

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B-cell CLL/Lymphoma 10 (BCL10) Is Required for NF-κB Production by Both Canonical and Noncanonical Pathways and for NF-κB-inducing Kinase (NIK) Phosphorylation

Bhattacharyya

Borthakur

Tyagi

et al. 2010

Journal of Biological Chemistry

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“…The most profound effects were observed with a dual inhibitor of both IRAKs, suggesting redundancy in the activities of the two enzymes. It has also been demonstrated that IRAK1/4 inhibitors result in a decrease in the production of interleukin-8 following LPS stimulation (22). We found that LPS-induced Mal degradation is also prevented upon inhibition of IRAK1/4.…”

Section: Discussionsupporting

confidence: 67%

IRAK1 and IRAK4 promote phosphorylation, ubiquitination, and degradation of MyD88 adaptor-like (Mal).

Dunne¹,

Carpenter²,

Brikos³

et al. 2016

Journal of Biological Chemistry

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Signal transduction by Toll-like receptor 2 (TLR2) and TLR4 requires the adaptors MyD88 and Mal (MyD88 adaptor-like) and serine/threonine kinases, interleukin-1 receptor-associated kinases IRAK1 and IRAK4. We have found that both IRAK1 and IRAK4 can directly phosphorylate Mal. In addition, co-expression of Mal with either IRAK resulted in depletion of Mal from cell lysates. This is likely to be due to Mal phosphorylation by the IRAKs because kinase-inactive forms of either IRAK had no effect. Furthermore, lipopolysaccharide stimulation resulted in ubiquitination and degradation of Mal, which was inhibited using an IRAK1/4 inhibitor or by knocking down expression of IRAK1 and IRAK4. MyD88 is not a substrate for either IRAK and did not undergo degradation. We therefore conclude that Mal is a substrate for IRAK1 and IRAK4 with phosphorylation promoting ubiquitination and degradation of Mal. This process may serve to negatively regulate signaling by TLR2 and TLR4.Activation of Toll-like receptor (TLR) 4 signaling pathways by microbial products initiates a cascade of events starting at the receptor level and leading eventually to the induction of an array of genes that encode for immune and inflammatory proteins. Ligand binding typically induces receptor dimerization, the recruitment of adaptor molecules, and the activation of a series of kinase cascades. In the case of the lipopolysaccharide (LPS) receptor TLR4 or the lipoprotein receptor TLR2, the adaptor molecules MyD88 (myeloid differentiation factor 88) and Mal (MyD88 adaptor-like) are recruited to activate NF-B and induce proinflammatory cytokines, such as interleukin-1, tumor necrosis factor ␣, and interleukin-6 (1-3). Two other adaptors, TRIF (Toll/interleukin-1 receptor domain-containing inducing interferon-␤) and TRAM (TRIF adaptor molecule), also participate in TLR4 signaling. Recruitment of these proteins leads to the activation of the transcription factor, IRF3 (interferon regulatory factor-3), and the induction of the type I interferons (4 -6).Mal and MyD88 both contain a Toll/interleukin-1 receptor (TIR) domain and function together to optimally transduce signals from TLR2 as well as TLR4. MyD88 differs from Mal in that it possesses an N-terminal death domain responsible for mediating interactions between MyD88 and members of the interleukin-1 receptor-associated kinase (IRAK) family. Four members of this family have been described, and the pathways leading to their activation have recently been dissected (7). In resting cells, IRAK1 can be found in a receptor complex containing MyD88 and Tollip. Following stimulation with LPS, IRAK1 is recruited to the TLR4 receptor complex, where it undergoes phosphorylation by IRAK4 on key threonine residues. This in turn promotes the autophosphorylating activity of IRAK1, its dissociation from the receptor complex, and subsequent interaction with TRAF6 (tumor necrosis factor receptor-associated factor) and a TAK1-TAB1-TAB2 kinase complex leading to the activation of NFB and mitogenactivated protein kinases. IRAK1...

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“…Secretion of IL-8 in the spent media of control and λCGN-treated NCM460 and primary colonic cells was measured using the DuoSet ELISA kit for human IL-8 (R&D Systems Inc, Minneapolis, MN), as described previously [5,9]. Sample values were normalized with total protein content (BCA™ protein assay; Pierce) and expressed as pg/mg protein.…”

Section: Measurement Of Il-8 Secretion By Elisamentioning

confidence: 99%

Carrageenan-induced NFκB activation depends on distinct pathways mediated by reactive oxygen species and Hsp27 or by Bcl10

Bhattacharyya

Dudeja

Tobacman

2008

Biochimica et Biophysica Acta (BBA) - General Subjects

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Carrageenans are highly sulfated polysaccharides that are widely used as food additives due to their ability to improve food texture. They are also widely recognized for their ability to induce inflammation in animal models of colitis. Recently, we reported that carrageenan (CGN) activated a pathway of innate immunity in human colonic epithelial cells mediated by Bcl10 (B-cell CLL/ lymphoma 10). However, increases in phospho-IκBα and Interleukin-8 (IL-8) were not completely inhibited by silencing Bcl10, suggesting that CGN also influenced another mechanism, or mechanisms, of inflammation. In this report, we demonstrate that CGN increases production of reactive oxygen species (ROS) in human colonic epithelial cells. The combination of ROS quenching by the free radical scavenger Tempol and of Bcl10 silencing by siRNA completely inhibited the CGN-induced increases in nuclear NFκB (p65), phospho-IκBα, and secretion of IL-8. The CGN-induced increase in ROS was associated with declines in phosphorylation of MAPK 12 (p38γ), MAPK 13 (p38δ), and heat-shock protein (Hsp) 27. The CGN-induced decline in phospho-Hsp27 was reversed by co-administration of Tempol (100nM), but unaffected by silencing Bcl10. Since Hsp27 phosphorylation is inversely associated with phosphorylation of the IκBα kinase (IKK) signalosome, CGN exposure appears to affect the IKK signalosome by both the catalytic component, mediated by ROS-phospho-Hsp27, and the regulatory component, mediated by Bcl10 interaction with IKKγ (Nemo). Hence, the CGN-activated inflammatory cascades related to innate immunity and to generation of ROS may be integrated at the level of the IKK signalosome.

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Bcl10 mediates LPS-induced activation of NF-κB and IL-8 in human intestinal epithelial cells

Cited by 42 publications

References 39 publications

B-cell CLL/Lymphoma 10 (BCL10) Is Required for NF-κB Production by Both Canonical and Noncanonical Pathways and for NF-κB-inducing Kinase (NIK) Phosphorylation

B-cell CLL/Lymphoma 10 (BCL10) Is Required for NF-κB Production by Both Canonical and Noncanonical Pathways and for NF-κB-inducing Kinase (NIK) Phosphorylation

IRAK1 and IRAK4 promote phosphorylation, ubiquitination, and degradation of MyD88 adaptor-like (Mal).

Carrageenan-induced NFκB activation depends on distinct pathways mediated by reactive oxygen species and Hsp27 or by Bcl10

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