NIK and IKKβ interdependence in NF-κB signalling—Flux analysis of regulation through metabolites

Kim, Hong-Bum; Evans, Iona; Smallwood, R. H.; Holcombe, Mike; Qwarnström, Eva E.

doi:10.1016/j.biosystems.2009.10.009

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…A minimum of 12 mutations can be observed for chickens with severe inflammatory cardiomyopathy. These mutations can skew coding regions of chromosomes, with subsequent functional alterations such as cell cycle regulation, clonal proliferation of immune system cells, and tissue injury (72,185,196,205,455). The complex intragenomic interactions affecting immune effectors and target cells may explain the variegated pathology in Chagas' disease.…”

Section: Use Of Tptail-pcr To Map Kdna Mutations In the Chicken Genomementioning

confidence: 99%

Pathogenesis of Chagas' Disease: Parasite Persistence and Autoimmunity

et al. 2011

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SUMMARY Acute Trypanosoma cruzi infections can be asymptomatic, but chronically infected individuals can die of Chagas' disease. The transfer of the parasite mitochondrial kinetoplast DNA (kDNA) minicircle to the genome of chagasic patients can explain the pathogenesis of the disease; in cases of Chagas' disease with evident cardiomyopathy, the kDNA minicircles integrate mainly into retrotransposons at several chromosomes, but the minicircles are also detected in coding regions of genes that regulate cell growth, differentiation, and immune responses. An accurate evaluation of the role played by the genotype alterations in the autoimmune rejection of self-tissues in Chagas' disease is achieved with the cross-kingdom chicken model system, which is refractory to T. cruzi infections. The inoculation of T. cruzi into embryonated eggs prior to incubation generates parasite-free chicks, which retain the kDNA minicircle sequence mainly in the macrochromosome coding genes. Crossbreeding transfers the kDNA mutations to the chicken progeny. The kDNA-mutated chickens develop severe cardiomyopathy in adult life and die of heart failure. The phenotyping of the lesions revealed that cytotoxic CD45, CD8 + γδ, and CD8α + T lymphocytes carry out the rejection of the chicken heart. These results suggest that the inflammatory cardiomyopathy of Chagas' disease is a genetically driven autoimmune disease.

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Section: Use Of Tptail-pcr To Map Kdna Mutations In the Chicken Genomementioning

confidence: 99%

Pathogenesis of Chagas' Disease: Parasite Persistence and Autoimmunity

et al. 2011

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“…Documentation of the kDNA minicircle integration in the chicken genome was obtained by a targeted prime TAIL-PCR, Southern hybridizations, cloning, and sequencing 3,4 . The kDNA minicircle integrations rupture open reading frames for transcription and immune system factors, phosphatase (GTPase), adenylate cyclase and phosphorylases (PKC, NF-Kappa B activator, PI-3K) associated with cell physiology, growth, and differentiation 3,[5][6][7] , and other gene functions. Severe myocarditis due to rejection of target heart fibers by effectors cytotoxic lymphocytes is seen in the kDNA mutated chickens, showing an inflammatory cardiomyopathy similar to that seen in human Chagas disease.…”

Section: Introductionmentioning

confidence: 99%

Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

Teixeira

Nitz

Bernal

et al. 2012

JoVE

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The Trypanosoma cruzi acute infections acquired in infancy and childhood seem asymptomatic, but approximately one third of the chronically infected cases show Chagas disease up to three decades or later. Autoimmunity and parasite persistence are competing theories to explain the pathogenesis of Chagas disease 1,2 . To separate roles played by parasite persistence and autoimmunity in Chagas disease we inoculate the T. cruzi in the air chamber of fertilized eggs. The mature chicken immune system is a tight biological barrier against T. cruzi and the infection is eradicated upon development of its immune system by the end of the first week of growth 3 . The chicks are parasite-free at hatching, but they retain integrated parasite mitochondrial kinetoplast DNA (kDNA) minicircle within their genome that are transferred to their progeny. Documentation of the kDNA minicircle integration in the chicken genome was obtained by a targeted prime TAIL-PCR, Southern hybridizations, cloning, and sequencing 3,4 . The kDNA minicircle integrations rupture open reading frames for transcription and immune system factors, phosphatase (GTPase), adenylate cyclase and phosphorylases (PKC, NF-Kappa B activator, PI-3K) associated with cell physiology, growth, and differentiation 3,[5][6][7] , and other gene functions. Severe myocarditis due to rejection of target heart fibers by effectors cytotoxic lymphocytes is seen in the kDNA mutated chickens, showing an inflammatory cardiomyopathy similar to that seen in human Chagas disease. Notably, heart failure and skeletal muscle weakness are present in adult chickens with kDNA rupture of the dystrophin gene in chromosome 1 8 . Similar genotipic alterations are associated with tissue destruction carried out by effectors CD45+, CD8γδ+, CD8α lymphocytes. Thus this protozoan infection can induce genetically driven autoimmune disease. Video LinkThe video component of this article can be found at https://www.jove.com/video/3716/ Protocol 1. Growth of Parasites 1. Grow trypomastigote forms of T. cruzi Berenice and the β-galactosidase-expressing Tulahuen T. cruzi MHOM/CH/00 C4 in murine muscle cell (L6) cultivated in Dulbecco minimal essential medium with 10% FSB, 100 IU/ml penicillin, 100 μg/ml streptomycin, and 250 nM L-glutamin (pH 7.2), 5% CO 2 at 37 °C. The free-swimming trypomastigotes in the supernatant medium were used to inoculate chicken eggs. 2. Grow Leishmania braziliensis (Lb) LTB300 stock grown in DMEM with 20% FBS. The Lb promastigote form in the exponential growth phase was used to inoculate eggs 9 . Parasite Inoculation in Fertilized Chicken Eggs1. Inoculate a suspension of 100 T. cruzi trypomastigotes in 10 μl of culture medium through a 2-mm diameter hole into the egg shell on top of the air chamber of stage X fertile eggs. The invasion and replication of the virulent parasites into the embryo cells are shown in Video S1. The control groups are as follows: a) control chickens; b) mock control eggs receiving 10 μl of culture medium; c) Stage X fertile eggs inoculated with a s...

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“…This proposition is further enforced by the fact that cells lacking both RelB and p52 subunits fail to generate meaningful osteoclasts when transduced with IKK2SSEE. In agreement with this phenomenon, several reports established cross-talk and interdependence of the classical and alternative NF-κB pathways in different responses [20]–[22]. In this regard, it has been shown that TNF, acting through the classical NF-κB pathway, up-regulates RelB expression in human umbilical vein endothelial cells [22].…”

Section: Discussionmentioning

confidence: 55%

Constitutively Active Canonical NF-κB Pathway Induces Severe Bone Loss in Mice

et al. 2012

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Physiologic osteoclastogenesis entails activation of multiple signal transduction pathways distal to the cell membrane receptor RANK. However, atypical osteoclastogenesis driven by pro-inflammatory stimuli has been described. We have reported recently a novel mechanism whereby endogenous mutational activation of the classical NF-κB pathway is sufficient to induce RANKL/RANK-independent osteoclastogenesis. Here we investigate the physiologic relevance of this phenomenon in vivo. Using a knock-in approach, the active form of IKK2, namely IKK2SSEE, was introduced into the myeloid lineage with the aid of CD11b-cre mice. Phenotypic assessment revealed that expression of IKK2SSEE in the myeloid compartment induced significant bone loss in vivo. This observation was supported by a dramatic increase in the number and size of osteoclasts in trabecular regions, elevated levels of circulating TRACP-5b, and reduced bone volume. Mechanistically, we observed that IKK2SSEE induced high expression of not only p65 but also p52 and RelB; the latter two molecules are considered exclusive members of the alternative NF-κB pathway. Intriguingly, RelB and P52 were both required to mediate the osteoclastogenic effect of IKK2SSEE and co-expression of these two proteins was sufficient to recapitulate osteoclastogenesis in the absence of RANKL or IKK2SSEE. Furthermore, we found that NF-κB2/p100 is a potent inhibitor of IKK2SSEE-induced osteoclastogenesis. Deletion of p52 enabled more robust osteoclast formation by the active kinase. In summary, molecular activation of IKK2 may play a role in conditions of pathologic bone destruction, which may be refractory to therapeutic interventions targeting the proximal RANKL/RANK signal.

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NIK and IKKβ interdependence in NF-κB signalling—Flux analysis of regulation through metabolites

Cited by 7 publications

References 25 publications

Pathogenesis of Chagas' Disease: Parasite Persistence and Autoimmunity

Pathogenesis of Chagas' Disease: Parasite Persistence and Autoimmunity

Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

Constitutively Active Canonical NF-κB Pathway Induces Severe Bone Loss in Mice

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