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Brijesh Kumar, John L Sorensen, Silvia T Cardona
Burkholderia cenocepacia is an opportunistic bacterium that can thrive in different environments, including the amino acid-rich mucus of the cystic fibrosis (CF) lung. B. cenocepacia responds to the nutritional conditions that mimic the CF sputum by increasing flagellin expression and swimming motility. Individual amino acids also induce swimming but not flagellin expression. Here, we show that modulation of the second messenger cyclic dimeric guanosine monophosphate (c-di-GMP) levels by the PAS-containing c-di-GMP phosphodiesterase, BCAL1069 (CdpA), regulates the swimming motility of B...
2018: Frontiers in Cellular and Infection Microbiology
Hwa Young Kim, Sang-Mi Yu, Sang Chul Jeong, Sang Sun Yoon, Young Taek Oh
The stringent response (SR), which is activated by accumulation of (p)ppGpp under conditions of growth-inhibiting stresses, plays an important role ongrowth and virulence in Vibrio cholerae . Herein, we carried out a genome-wide screen using transposon random mutagenesis to identify genes controlled by SR in (p)ppGpp-overproducing mutant strain. One of the identified SR target genes was the flaC encoding flagellin. Genetic studies using flaC and SR mutants demonstrated that FlaC involved in bacterial growth, toxin production, and normal flagellum function under conditions of high (p)ppGpp levels, suggesting FlaC plays an important role in SR-induced pathogenicityin V...
March 15, 2018: Journal of Microbiology and Biotechnology
Manoochehr Makvandi, Ali Teimoori, Mehdi Parsa Nahad, Ali Khodadadi, Mohammad Ghasemi Deh Cheshmeh, Milad Zandi
BACKGROUND: Flagellin is the major structural protein monomer of bacterial flagella. Flagellin through binding to its receptor and activation of antigen presenting cells stimulates the innate and adaptive immune responses. Flagellin is used as an effective systemic or mucosal adjuvant to stimulate the immune system. Recently, the therapeutic and protective role of flagellin in some infectious diseases and cancers has been investigated. In this study, we cloned the fliC genes from Salmonella typhimurium and Escherichia coli into pET-28a vector and investigated their expression in the prokaryotic system...
March 9, 2018: Microbial Pathogenesis
M M Vaezirad, M G Koene, J A Wagenaar, J P M van Putten
In ovo immunization of chicken embryos with live vaccines is an effective strategy to protect chickens against several viral pathogens. We investigated the immune response of chicken embryos to purified recombinant protein. In ovo delivery of Salmonella flagellin to 18-day old embryonated eggs resulted in elevated pro-inflammatory chIL-6 and chIL-8 (CXCL8-CXCLi2) cytokine transcript levels in the intestine but not in the spleen at 24 h post-injection. Analysis of the chicken Toll-like receptor (TLR) repertoire in 19-day old embryos revealed gene transcripts in intestinal and spleen tissue for most chicken TLRs, including TLR5 which recognizes Salmonella flagellin (FliC)...
March 9, 2018: Vaccine
Kelly Flentie, Caleb Gonzalez, Brandon Kocher, Yue Wang, Hongtu Zhu, Jayne Marasa, David Piwnica-Worms
Bacterial flagellin is a potent activator of NF-κB signaling, inflammation and host innate immunity, and recent data indicate that flagellin represents a novel anti-tumor ligand acting through toll-like receptor 5 (TLR5) and the NF-κB pathway to induce host immunity and aid in the clearance of tumor xenografts. To identify innate signaling components of TLR5 responsible for these anti-tumor effects, a loss-of-function high-throughput screen was employed utilizing carcinoma cells expressing a dynamic NF-κB bioluminescent reporter stimulated by Salmonella typhimurium expressing flagellin...
March 9, 2018: Molecular Cancer Research: MCR
Eugenia S Mardanova, Nikolai V Ravin
BACKGROUND: Influenza is a widely distributed infection that almost annually causes seasonal epidemics. The current egg-based platforms for influenza vaccine production are facing a number of challenges and are failing to satisfy the global demand in the case of pandemics due to the long production time. Recombinant vaccines are an alternative that can be quickly produced in high quantities in standard expression systems. METHODS: Plants may become a promising biofactory for the large-scale production of recombinant proteins due to low cost, scalability, and safety...
March 9, 2018: Current Pharmaceutical Design
Sana Syed, Najeeha T Iqbal, Kamran Sadiq, Jennie Z Ma, Tauseef Akhund, Wenjun Xin, Sean R Moore, Enju Liu, Shahida Qureshi, Kerri Gosselin, Andrew Gewirtz, Christopher P Duggan, S Asad Ali
BACKGROUND: Environmental Enteric Dysfunction (EED) in children from low-income countries has been linked to linear growth declines. There is a critical need to identify sensitive and early EED biomarkers. OBJECTIVE: Determine whether levels of antibodies against bacterial components flagellin (flic) and lipopolysaccharide (LPS) predict poor growth. DESIGN/METHODS: In a prospective birth cohort of 380 children in rural Pakistan blood and stool samples were obtained at ages 6 and 9 months...
2018: PloS One
Lara Ajamian, Luca Melnychuk, Patrick Jean-Pierre, Gerasimos J Zaharatos
Flagellin's potential as a vaccine adjuvant has been increasingly explored over the last three decades. Monomeric flagellin proteins are the only known agonists of Toll-like receptor 5 (TLR5). This interaction evokes a pro-inflammatory state that impacts upon both innate and adaptive immunity. While pathogen associated molecular patterns (PAMPs) like flagellin have been used as stand-alone adjuvants that are co-delivered with antigen, some investigators have demonstrated a distinct advantage to incorporating antigen epitopes within the structure of flagellin itself...
February 27, 2018: Viruses
Yoshiko Kudo, Ket Vansith, En Rin, Kohdai Uchida, Sumika Kodama, Takashi Fukui, Sho Masuda, Toshiyuki Masuzawa
INTRODUCTION: Leptospirosis remains a major public health threat in Cambodia. In this study, we aimed at facilitating the development of preventive strategies against leptospirosis in Cambodia by conducting molecular epidemiological surveys of Leptospira infection among wild rodents. MATERIALS AND METHODS: One hundred sixty-three wild rodents were captured in the capital Phnom Penh and surrounding areas, and their kidneys and bladders were collected for analysis...
March 2018: Vector Borne and Zoonotic Diseases
Gwladys Sevrin, Sébastien Massier, Benoit Chassaing, Allison Agus, Julien Delmas, Jérémy Denizot, Elisabeth Billard, Nicolas Barnich
The pathogenesis of Crohn's disease (CD) is multifactorial and involves genetic susceptibility, environmental triggers and intestinal microbiota. Adherent-invasive Escherichia coli (AIEC) are flagellated bacteria more prevalent in CD patients than in healthy subjects and promote chronic intestinal inflammation. We aim at deciphering the role of flagella and flagellin modulation by intestinal conditions. AIEC flagellum expression is required for optimal adhesion to and invasion of intestinal epithelial cells...
March 1, 2018: Gut Microbes
Bettina L Lee, Kathleen M Mirrashidi, Irma B Stowe, Sarah K Kummerfeld, Colin Watanabe, Benjamin Haley, Trinna L Cuellar, Michael Reichelt, Nobuhiko Kayagaki
The NLRC4 inflammasome recognizes bacterial flagellin and components of the type III secretion apparatus. NLRC4 stimulation leads to caspase-1 activation followed by a rapid lytic cell death known as pyroptosis. NLRC4 is linked to pathogen-free auto-inflammatory diseases, suggesting a role for NLRC4 in sterile inflammation. Here, we show that NLRC4 activates an alternative cell death program morphologically similar to apoptosis in caspase-1-deficient BMDMs. By performing an unbiased genome-wide CRISPR/Cas9 screen with subsequent validation studies in gene-targeted mice, we highlight a critical role for caspase-8 and ASC adaptor in an alternative apoptotic pathway downstream of NLRC4...
February 28, 2018: Scientific Reports
Ágnes Klein, Mátyás Kovács, Adél Muskotál, Hajnalka Jankovics, Balázs Tóth, Mihály Pósfai, Ferenc Vonderviszt
In this work we addressed the problem how to fabricate self-assembling tubular nanostructures displaying target recognition functionalities. Bacterial flagellar filaments, composed of thousands of flagellin subunits, were used as scaffolds to display single-domain antibodies (nanobodies) on their surface. As a representative example, an anti-GFP nanobody was successfully inserted into the middle part of flagellin replacing the hypervariable surface-exposed D3 domain. A novel procedure was developed to select appropriate linkers required for functional internal insertion...
February 26, 2018: Scientific Reports
Dongming Wu, Rong Han, Shihua Deng, Teng Liu, Ting Zhang, Hongxiang Xie, Ying Xu
PURPOSE: To investigate changes induced in mouse intestines after irradiation and to explore the potential radioprotective effects of flagellin A N/C (FlaAN/C). METHODS AND MATERIALS: A mouse model of radiation-induced enteropathy was used in this study. A 10-Gy abdominal irradiation was performed on FlaAN/C- and vehicle-injected mice to explore the role of FlaAN/C in intestinal radiation injury and to study the molecular mechanism in this process. In the intestinal tissue, pathologic changes were investigated by hematoxylin and eosin staining, TUNEL staining, and immunohistochemistry; Western blotting and quantitative reverse transcription-polymerase chain reaction were used to determine the changes in protein and messenger RNA levels, respectively; and an enzyme-linked immunosorbent assay was performed to determine protein concentration in serum...
January 31, 2018: International Journal of Radiation Oncology, Biology, Physics
Mehri Habibi, Mohammad Reza Asadi Karam, Saeid Bouzari
Urinary tract infections (UTI) caused by Proteus mirabilis are prevalent among the catheterized patients. There is no effective vaccine to reduce the frequency of UTIs caused by P. mirabilis. In the present study, the immune responses and effectiveness of different combinations of MrpA and flagellin (FliC) of P. mirabilis were assessed intranasally in the mice model. The addition of FliC as adjuvant to MrpA in fusion form significantly raised the mucosal IgA and cellular (IFN-γ and IL-17) responses and maintained the serum IgG responses for 180 days after the first vaccination...
February 13, 2018: Microbial Pathogenesis
Gayeon Won, John Hwa Lee
The obligate intracellular Lawsonia intracellularis (LI), the etiological agent of proliferative enteropathy (PE), is an economically important disease in the swine industry. Due to extreme difficulty of in vitro culture of the pathogen, molecular characterization of protein components of LI that are targets of the immune system, is difficult; thus, the scientific evidence to drive the development of preventive measures is lacking. In this work, we investigated the antigenic and functional characteristics of a putative flagellar-associated protein, LI0570, using in silico computational approaches for epitope prediction and an in vitro protein-based molecular assay...
February 15, 2018: Veterinary Research
Melanie K B Wills, Andrea M Kirby, Vett K Lloyd
Lyme disease is a serious vector-borne infection that is caused by the Borrelia burgdorferi sensu lato family of spirochetes, which are transmitted to humans through the bite of infected Ixodes ticks. The primary etiological agent in North America is Borrelia burgdorferi sensu stricto. As geographic risk regions expand, it is prudent to support robust surveillance programs that can measure tick infection rates, and communicate findings to clinicians, veterinarians, and the general public. The molecular technique of nested polymerase chain reaction (nPCR) has long been used for this purpose, and it remains a central, inexpensive, and robust approach in the detection of Borrelia in both ticks and wildlife...
February 4, 2018: Journal of Visualized Experiments: JoVE
Haojie Yan, Yaofei Zhao, Hua Shi, Juan Li, Yingchun Wang, Dingzhong Tang
Arabidopsis immune receptor FLAGELLIN SENSING2 (FLS2) rapidly forms a complex to activate pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) upon perception of the bacterial protein flagellin. The receptor-like cytoplasmic kinase BRASSINOSTEROID-SIGNALINGKINASE1 (BSK1) interacts with FLS2 and is critical for the activation of PTI. However, it is unknown how BSK1 transduces signals to activate downstream immune responses. We identified Mitogen-Activated Protein Kinase Kinase Kinase5 (MAPKKK5) as a potential substrate of BSK1 by whole-genome phosphorylation analysis...
February 12, 2018: Plant Physiology
Keke Shang-Guan, Min Wang, Nang Myint Phyu Sin Htwe, Ping Li, Yaoshen Li, Fan Qi, Dawei Zhang, Min Cao, Chanhong Kim, Haiyong Weng, Haiyan Cen, Ian M Black, Parastoo Azadi, Russell W Carlson, Gary Stacey, Yan Liang
Lipopolysaccharide (LPS), a major component of the gram-negative bacterial outer membrane, is an important microbe-associated molecular pattern (MAMP) that triggers immune responses in both plants and animals. In Arabidopsis, LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (LORE), a B-type lectin S-domain receptor kinase, was previously identified as an LPS sensor through a genetic screen. However, the LPS-activated LORE signaling pathway and associated immune responses are still largely unknown. In this study, we found that LPS triggered a biphasic production of reactive oxygen species (ROS) in Arabidopsis...
February 5, 2018: Plant Physiology
Ratana Lim, Martha Lappas
PROBLEM: Inflammation plays a major role in preterm birth. Nucleotide-binding oligomerization domain-like receptor pyrin domain-containing-3 (NLRP3) plays a role in inflammatory diseases. The aims of this study were to determine the effect of term labor on the expression of NLRP3 in human myometrium and the effect of NLRP3 silencing on pro-labor mediators in myometrial cells. METHOD OF STUDY: NLRP3 expression was assessed in myometrium from non-laboring and laboring women by qRT-PCR and Western blotting...
February 12, 2018: American Journal of Reproductive Immunology: AJRI
Qianniu Li, Ouyang Peng, Tingting Wu, Zhichao Xu, Licheng Huang, Yun Zhang, Chunyi Xue, Zhifen Wen, Qingfeng Zhou, Yongchang Cao
Porcine epidemic diarrhea (PED) is an important re-emergent infectious disease and inflicts huge economic losses to the swine industry worldwide. To meet the pressing need of developing a safe and cost-efficient PED maternal vaccine, we generated three PED subunit vaccine candidates, using recombined Salmonella flagellin (rSF) as a mucosal molecular adjuvant. Domain D3 in rSF was replaced with COE domain of PEDV to generate rSF-COE-3D. COE fused to the flanking C'/N' terminal of rSF yielded rSF-COE-C and rSF-COE-N...
February 6, 2018: Vaccine
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