RESUMO
The cellular oncogene c-Fos (c-Fos) is a component of activator protein 1 (AP1), a master transcriptional regulator of cells. The suppression of c-Fos signaling by siRNA treatment resulted in significant induction of TLR4, which subsequently activates p38 and ERK1/2 mitogen-activated protein kinases (MAPKs) and enhances F-actin polymerization, leading to an increase in B. abortus phagocytosis. During B. abortus infection, c-Fos signaling is induced, which activates the downstream innate-immunity signaling cascade for bacterial clearance. The inhibition of c-Fos signaling led to increased production of interleukin 10 (IL-10), which partially suppressed lysosome-mediated killing, resulting in increased survival of B. abortus inside macrophages. We present evidence of the regulatory role played by the c-Fos pathway in proliferation during B. abortus infection; however, this was independent of the anti-Brucella effect of this pathway. Another finding is the essential contribution of c-Fos/TRAIL to infected-cell necrosis, which is a key event in bacterial dissemination. These data provide the mechanism via which c-Fos participates in host defense mechanisms against Brucella infection and in bacterial dissemination by macrophages.
Assuntos
Brucella/crescimento & desenvolvimento , Brucella/imunologia , Interações Hospedeiro-Patógeno , Macrófagos/imunologia , Macrófagos/microbiologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Sobrevivência Celular , Camundongos , Células RAW 264.7RESUMO
In this study, we explore the regulatory roles of pro-inflammatory cytokine tumor necrosis factor alpha (TNF) in the innate immunity of macrophages against B. abortus infection. We show that infection of macrophage with B. abortus induces marked expression and secretion of TNF which subsequently binds to TNF receptor 1 (TNFR-1) and activates a downstream signaling cascade of the innate immunity. Blocking of TNF signaling resulted in a notable increase of B. abortus survival which was associated with an increase of anti-inflammatory cytokine interleukin 10 (IL-10), a beneficial effector of Brucella survival, as well as remarkable decrease of reactive oxygen species (ROS) and nitric oxide (NO), antibrucella molecules. However, surprisingly, the interference of TNF did not show any influence on phagolysosome and cell death events. Furthermore, the transcriptional factor NF-kB was found to be a main mediator of TNF signaling when blocking of NF-kB pathway drastically suppressed the TNF-induced brucellacidal effect. Taken together, these findings clearly indicate that the immune cascade activated by TNF/TNFR-1 is required for the sufficient resistance to B. abortus survival in macrophages.
Assuntos
Brucella abortus/imunologia , Brucelose/imunologia , Imunidade Inata , NF-kappa B/imunologia , Receptores Tipo I de Fatores de Necrose Tumoral/imunologia , Animais , Morte Celular/imunologia , Humanos , Interleucina-10/imunologia , Camundongos , Óxido Nítrico/imunologia , Fagossomos/imunologia , Células RAW 264.7 , Espécies Reativas de Oxigênio/imunologiaRESUMO
Interleukin-17 (IL17), belonging to the Th17 family, is a proinflammatory cytokine produced by activated T cells. A 1034bp cDNA encoding duck IL17 (duIL17) was cloned from Con A-activated splenic lymphocytes of ducks. The encoded protein, which is predicted to consist of 169 amino acids, has a molecular weight of 18.8kDa and includes a 29 residue NH(2)-terminal signal peptide, a single potential N-linked glycosylation site, and six cysteine residues that are conserved in mammalian IL17. The duIL17 shared 84% amino acid sequence identity with the previously described chicken IL17 (chIL17), 36-47% to mammalian homologues, and open reading frame 13 of Herpesvirus saimiri (HVS13). The genomic structure of duIL17 was quite similar to its chicken and mammalian counterparts. The duIL17 mRNA expression was detected only in Con A-activated splenic lymphocytes by RT-PCR, although its expression was undetectable in a variety of normal tissues. Two mAbs against chIL17 showed cross-reactivity with duIL17 as detected by indirect ELISA and Western blot analysis. These findings indicate that the structure of IL17 is highly conserved among poultry, and two mAbs detecting common epitopes of IL17 are available for molecular and immunological studies of IL17 in birds.