Brucella melitensis is a facultative intracellular bacterium that causes brucellosis, the most prevalent zoonosis worldwide. The replicative niche in macrophages and dendritic cells thwarts immune surveillance complicates both therapy vaccine development. Currently, host-pathogen interactions supporting replication are poorly understood. fuses with endoplasmic reticulum (ER) to replicate, resulting dramatic restructuring of ER. This ER disruption raises possibility provokes an stress...

The use of mammalian gene expression vectors has become increasingly important for transgenics and therapy as well basic research. Essential the success these in medical research applications is proper choice promoter linked to interest. Many elements from pathogenic viruses, including simian virus 40 (SV40) cytomegalovirus (CMV). Lymphokines produced by immune response proteins expressed could inhibit further transcription initiation viral promoters. Our objective was determine effect...

Abstract Immunity against microbes depends on recognition of pathogen-associated molecular patterns by innate receptors. Signaling pathways triggered Brucella abortus DNA involves TLR9, AIM2, and stimulator IFN genes (STING). In this study, we observed microarray analysis that several type I IFN–associated genes, such as IFN-β guanylate-binding proteins (GBPs), are downregulated in STING knockout (KO) macrophages infected with or transfected DNA. Additionally, determined cyclic GMP–AMP...

Brucella abortus is a facultative intracellular bacterium that causes brucellosis, prevalent zoonosis leads to abortion and infertility in cattle, undulant fever, debilitating arthritis, endocarditis, meningitis humans. Signaling pathways triggered by B. involves stimulator of IFN genes (STING), which production type I IFNs. In this study, we evaluated the pathway linking unfolded protein response (UPR) endoplasmic reticulum-resident transmembrane molecule STING, during infection. We...

Brucellosis is a common zoonotic disease that remains endemic in many parts of the world. Dissecting host immune response during this provides insight as to why brucellosis often difficult resolve. We used Brucella epitope specific vivo killing assay investigate ability CD8+ T cells kill targets treated with purified pathogenic protein. Importantly, we found protein TcpB be novel effector adaptive evasion by inhibiting cell target mice. Further, BALB/c mice show active melitensis infection...

TIR (Toll/interleukin-1 receptor) domain-containing proteins play a crucial role in innate immunity eukaryotes. Brucella is highly infectious intracellular bacterium that encodes domain protein (TcpB) to subvert host immune responses establish beneficial niche for pathogenesis. TcpB inhibits NF-κB (nuclear factor κB) activation and pro-inflammatory cytokine secretions mediated by TLR (Toll-like 2 TLR4. In the present study, we have demonstrated modulates microtubule dynamics acting as...

Abstract Brucellosis, one of the most prevalent zoonotic diseases worldwide, often results in osteoarticular complications including large joint and axial arthritis mimicking spondyloarthritis. To model this chronic manifestation, we infected autoimmunity-prone SKG mice containing a mutation T-cell adaptor ZAP-70 with Brucella species. B. melitensis infection resulted fully penetrant, readily scoreable disease involving wrist foot arthritis, peri-ocular inflammation, less frequent scaly paw...

ABSTRACT The alphaherpesvirus tegument protein VP22 has been characterized with multiple traits including microtubule reorganization, nuclear localization, and nonclassical intercellular trafficking. However, all these data were derived from studies using herpes simplex virus type 1 (HSV-1) may not apply to homologs of other alphaherpesviruses. We compared subcellular attributes HSV-1 (HVP22) bovine herpesvirus (BHV-1) (BVP22) green fluorescent (GFP)-fused expression vectors. Fluorescence...

Brucella spp. are gram-negative bacteria that cause the most frequent zoonotic disease worldwide, with more than 500,000 human infections yearly; however, no vaccine is currently available. As other intracellular organisms, cytotoxic mechanisms against infected cells thought to have an important role in controlling infection and mediating long-term immunity. Live attenuated strains developed for use animals elicit protection but retain unacceptable levels of virulence. Thus, optimal design a...

ABSTRACT Brucella melitensis encounters a variety of conditions and stimuli during its life cycle—including environmental growth, intracellular infection, extracellular dissemination—which necessitates flexibility bacterial signaling to promote virulence. Cyclic-di-GMP is secondary molecule that plays an important role in adaptation changing environments altering virulence number bacteria. To investigate the cyclic-di-GMP B. , all 11 predicted cyclic-di-GMP-metabolizing proteins were...

Brucella melitensis is a well-adapted zoonotic pathogen considered scourge of mankind since recorded history. In some cases, initial infection leads to chronic and reactivating brucellosis, incurring significant morbidity economic loss. The mechanism by which B. subverts adaptive immunological memory poorly understood. Previous work has shown that Brucella-specific CD8(+) T cells express gamma interferon (IFN-γ) can transition long-lived but are not polyfunctional. this study, mice with led...

The immunogenicity and protective efficacy of a DNA vaccine encoding the GroEL heat-shock gene from Brucella abortus was tested in BALB/c mice immunised by intramuscular (i.m.) needle injection or epidermally gun. amplified PCR cloned into two different mammalian expression vectors pCMV-link pCMV-tPA. D17 cell line transfected with both constructs transcripts were detected Northern blot. To determine level protein synthesised, lysates then submitted to Western non-secreted form recombinant...

Cellular immune responses are a critical part of the host's defense against intracellular bacterial infections. Immunity to Brucella abortus crucially depends on antigen-specific T cell-mediated activation macrophages, which major effectors killing this organism. lymphocytes that proliferate in response B. were characterized for phenotype and cytokine activity. Human, murine, bovine exhibited type 1 profile, suggesting an analogous these different hosts. In vivo protection afforded by...

Carboxyfluorescein diacetate succinimidyl ester (CFSE) can be used to easily and quickly label a cell population of interest for in vivo investigation. This labeling has classically been study proliferation migration. In the method presented here, we have shortened timeline after adoptive transfer look at survival killing epitope specific CFSE labeled target cells. The level CD8 + T clone indicate quality response, as their quantity may misleading. Specific CD8+ cells become functionally...

Brucellosis, caused by a number of Brucella species, remains the most prevalent zoonotic disease worldwide. establish chronic infections within host macrophages despite triggering cytosolic innate immune sensors, including Stimulator Interferon Genes (STING), which potentially limit infection. In this study, STING was required for control infection in vivo . However, early during infection, down-regulated mRNA and protein. Down-regulation occurred post-transcriptionally, live bacteria, type...

Brucella species are facultative intracellular bacteria that cause brucellosis, a chronic debilitating disease significantly impacting global health and prosperity. Much remains to be learned about how spp. succeed in sabotaging immune host cells respond environmental challenges. Multiple types of employ the prokaryotic second messenger cyclic di-GMP (c-di-GMP) coordinate responses shifting environments. To determine role c-di-GMP physiology shaping host-Brucella interactions, we utilized...

ABSTRACT The bovine herpesvirus 1 (BHV-1) UL49 gene encodes a viral tegument protein termed VP22. homologs are conserved among alphaherpesviruses. Interestingly, the BHV-1 VP22 deletion mutant virus is asymptomatic and avirulent in infected cattle but produces only slight reduction titer vitro. Attenuation of vivo suggests that plays functional role replication. In herpes simplex type 1, homolog was previously shown to interact with another protein,VP16, α-transinducing factor this report,...

Brucella spp. are intracellular bacteria that cause the most frequent zoonosis in world. Although recent work has advanced field of vaccine development, there remains no safe human vaccine. In order to produce a and effective vaccine, immune response requires greater understanding. Induction Brucella-specific CD8+ T cells is considered an important aspect host response; however, T-cell not clearly defined. Discovering epitope containing antigens recognized by correlating them with microarray...

There is no safe, effective human vaccine against brucellosis. Live attenuated Brucella strains are widely used to vaccinate animals. However these live vaccines can cause disease and unsafe for humans. Killed or subunit not in eliciting long term protection. In this study, we evaluate an approach using a live, non-pathogenic bacteria (E. coli) genetically engineered mimic the brucellae pathway of infection present antigens appropriate cytolitic T cell response.E. coli was modified express...