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Kartik Ramakrishna, Kalpana Premkumar, Jayakanthan Kabeerdoss, K R John
BACKGROUND & AIM: Innate immune responses are important in susceptibility to pulmonary tuberculosis (TB). In order to test the hypothesis that Toll-like receptor (TLR) 2 function would be abnormal in patients with active pulmonary TB we compared the cytokine responses of peripheral blood mononuclear cells (PBMC) to innate immune ligands in a case-control study. METHODS: PBMC from 19 untreated pulmonary TB patients, 17 healthy controls, and 11 treated pulmonary TB patients, were cultured for 24h with TLR 2 ligand (PAM-CSK) and other TLR ligands (muramyl dipeptide, flagellin, lipopolysaccharide (LPS), CpG oligodeoxynucleotide (CpG-ODN))...
October 18, 2016: Cytokine
Katarzyna A Radomska, Mahdi M Vaezirad, Koen M Verstappen, Marc M S M Wösten, Jaap A Wagenaar, Jos P M van Putten
Campylobacter jejuni is the main cause of bacterial food-borne diseases in developed countries. Chickens are the most important source of human infection. Vaccination of poultry is an attractive strategy to reduce the number of C. jejuni in the intestinal tract of chickens. We investigated the immunogenicity and protective efficacy of a recombinant C. jejuni flagellin-based subunit vaccine with intrinsic adjuvant activity. Toll-like receptor activation assays demonstrated the purity and TLR5 stimulating (adjuvant) activity of the vaccine...
2016: PloS One
Laura Bouché, Maria Panico, Paul Hitchen, Daniel Binet, Federico Sastre, Alexandra Faulds-Pain, Esmeralda Valiente, Evgeny Vinogradov, Annie Aubry, Kelly Fulton, Susan Twine, Susan M Logan, Brendan W Wren, Anne Dell, Howard R Morris
Glycosylation of flagellins is a well recognized property of many bacterial species. In this study we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile. We used mass spectrometry (nano LC- MS and MS/MS analysis) to identify a number of putative glycopeptides which carried a variety of glycoform substitutions each of which was linked through an initial HexNAc residue to Ser or Thr. Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity...
October 7, 2016: Journal of Biological Chemistry
Michelle E Leslie, Sean W Rogers, Antje Heese
Callose deposition within the cell wall is a well-documented plant immune response to pathogenic organisms as well as to pathogen-/microbe- associated molecular patterns (P/MAMPs). However, the molecular mechanisms that modulate pathogen-induced callose deposition are less understood. We reported previously that Arabidopsis plants lacking the vesicle trafficking component DYNAMIN-RELATED PROTEIN 2B (DRP2B) display increased callose deposition in response to the PAMP flg22. Here, we show that increased number of flg22-induced callose deposits in drp2b leaves is fully dependent on the callose synthase POWDERY MILDEW RESISTANT 4 (PMR4)...
October 17, 2016: Plant Signaling & Behavior
Lei Li, Panya Kim, Liping Yu, Gaihong Cai, She Chen, James R Alfano, Jian-Min Zhou
The Arabidopsis immune receptor FLS2 and co-receptor BAK1 perceive the bacterial flagellin epitope flg22 to activate plant immunity. To prevent this response, phytopathogenic bacteria deploy a repertoire of effector proteins to perturb immune signaling. However, the effector-induced perturbation is often sensed by the host, triggering another layer of immunity. We report that the Pseudomonas syringae effector HopB1 acts as a protease to cleave immune-activated BAK1. Prior to activation, HopB1 constitutively interacts with FLS2...
October 12, 2016: Cell Host & Microbe
Xilong Kang, Yun Yang, Yang Jiao, Hongqin Song, Li Song, Dan Xiong, Lili Wu, Zhiming Pan, Xinan Jiao
In 2009, a novel pandemic swine-origin influenza A (H1N1) virus caused a public emergency of international concern. Vaccination is the primary strategy for the control of influenza epidemics. However, the poor immunopotency of many vaccine antigens is a major barrier to the development of effective vaccines against influenza. Flagellin, a Toll-like receptor 5 (TLR5) ligand, has been used as an adjuvant to enhance the immunopotency of vaccines in preclinical studies. Here, we developed a recombinant candidate vaccine, HA1-2-fljB, in which the globular head of the hemagglutinin (HA) antigen (residues 62-284) from H1N1 virus was fused genetically to the N-terminus of Salmonella typhimurium fljB...
October 8, 2016: Journal of Molecular Microbiology and Biotechnology
Atsushi Kimishima, Cody J Wenthur, Lisa M Eubanks, Shun Sato, Kim D Janda
Although cocaine abuse and addiction continue to cause serious health and societal problems, an FDA-approved medication to treat cocaine addiction has yet to be developed. Employing a pharmacokinetic strategy, an anti-cocaine vaccine, provides an attractive avenue to address these issues, however, current vaccines have shown varying degrees of efficacy, indicating that further formulation is necessary. As a means to improve vaccine efficacy, we examined the effects of varying anti-cocaine vaccine formulations by combining a Toll-like receptor 9 (TLR9) agonist with a TLR5 agonist in the presence of alum...
October 7, 2016: Molecular Pharmaceutics
Dominika Drzewiecka, Gabriela Lewandowska
Urinary tract infections (UTIs) pose a threat especially to women, the individuals with weakened immunity or with abnormalities in the urinary tract as well as to hospitalized and catheterized patients. The bacteria from the genus Proteus, especially P. mirabilis, are important UTI pathogenic factors. They frequently cause chronic, recurrent or severely complicated infections, resulting in the urinary stones production due to urease and other virulence factors. The ability to survive inside the stones and the increasing antibiotic resistance make it difficult to eradicate the bacteria from the urinary tract...
September 30, 2016: Postȩpy Higieny i Medycyny Doświadczalnej
Ying Xu, Dongming Wu, Yuanchun Fan, Peigeng Li, Hongfei Du, Jiao Shi, Dan Wang, Xiaoping Zhou
The recombinant protein flagellin A (FlaA) N/C, derived from the flagellin protein of Legionella pneumophila, has been shown to increase the expression of cytoprotective cytokines, activate the nuclear factor-κB (NF-κB) signaling pathway, and increase the survival of mice following total body irradiation. Determi ning whether FlaA N/C has a sensitizing effect on tumor radiation or a direct tumoricidal effect is critical for its application as an effective radiation protection agent. The present study investigated the molecular mechanism underlying the tumor radiosensitivity of FlaA N/C...
October 2016: Oncology Letters
Esmeralda Valiente, Laura Bouché, Paul Hitchen, Alexandra Faulds-Pain, Mario Songane, Lisa F Dawson, Elizabeth Donahue, Richard A Stabler, Maria Panico, Howard R Morris, Mona Bajaj-Elliott, Susan M Logan, Anne Dell, Brendan W Wren
Clostridium difficile is the principal cause of nosocomial infectious diarrhoea worldwide. The pathogen modifies its flagellin with either a type A or type B O-linked glycosylation system, which has a contributory role in pathogenesis. We study the functional role of glycosyltransferases modifying type B flagellin in the 023 and 027 hypervirulent C. difficile lineages by mutagenesis of five putative glycosyltransferases and biosynthetic genes. We reveal their roles in the biosynthesis of the flagellin glycan chain and demonstrate that flagellar post-translational modification affects motility and adhesion-related bacterial properties of these strains...
October 4, 2016: Journal of Biological Chemistry
Tanujaa Suriyanarayanan, Saravanan Periasamy, Miao-Hsia Lin, Yasushi Ishihama, Sanjay Swarup
Protein phosphorylation has a major role in controlling the life-cycle and infection stages of bacteria. Proteome-wide occurrence of S/T/Y phosphorylation has been reported for many prokaryotic systems. Previously, we reported the phosphoproteome of Pseudomonas aeruginosa and Pseudomonas putida. In this study, we show the role of S/T phosphorylation of one motility protein, FliC, in regulating multiple surface-associated phenomena of P. aeruginosa PAO1. This is the first report of occurrence of phosphorylation in the flagellar protein, flagellin FliC in its highly conserved N-terminal NDO domain across several Gram negative bacteria...
2016: PloS One
Qiong Liu, Qing Liu, Jie Yi, Kang Liang, Bo Hu, Xiangmin Zhang, Roy Curtiss, Qingke Kong
Outer membrane vesicles (OMVs) isolated from Salmonella Typhimurium are potentially useful for developing subunit vaccines because of high immunogenicity and protective efficacy. However, flagella might remain in OMV pellets following OMV purification, resulting in non-essential immune responses and counteraction of bacterial protective immune responses when developing a vaccine against infection of multiple serotypes Salmonella. In this study, a flagellin-deficient S. Typhimurium mutant was constructed. Lipopolysaccharide profiles, protein profiles and cryo-electron microscopy revealed that there were no significant differences between the wild-type and mutant OMVs, with the exception of a large amount of flagellin in the wild-type OMVs...
October 4, 2016: Scientific Reports
Kurni Kurniyati, John F Kelly, Evgeny Vinogradov, Anna Robotham, Youbing Tu, Juyu Wang, Jun Liu, Susan M Logan, Chunhao Li
While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. Herein, we report a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis. The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues...
October 1, 2016: Molecular Microbiology
Mozhgan Derakhshan Sefidi, Iraj Rasooli, Parviz Owlia, Daryush Talei, Shakiba Darvish Alipour Astaneh, Shahram Nazarian
AIM: To study immunogenicity of Pseudomonas N terminal flagellin as an adjuvant for Acinetobacter baumannii (A. baumannii) biofilm associated protein (Bap). METHODS: The N terminal flagellin gene was amplified. The pET28a (+) and polymerase chain reaction products were digested with HindIII and EcoR I. The ligation of N terminal flagellin into pET28a (‏+) was performed using T4 DNA ligase and was then transformed into Escherichia coli BL21 (DE3) as a suitable expression host...
September 26, 2016: World Journal of Methodology
Yang Xu, Hongxia Dong, Changhui Ge, Yan Gao, Haifeng Liu, Weiguang Li, Chenggang Zhang
BACKGROUND: Ulcerative colitis (UC) is a nonspecific inflammatory disease for which medications and therapeutic strategies have only been moderately successful. CBLB502, a toll-like receptor 5 (TLR5) agonist derived from Salmonella flagellin, exhibits anticancer and radioprotective activities via modulation of TLRs and the nuclear factor kappa B (NF-κB) signaling pathway and can protect against acute renal ischemic failure. In this study, we intend to examine the effects of CBLB502 on both TLR responses and the interleukin (IL) and NF-κB signaling pathways in UC treatment...
August 2016: Annals of Translational Medicine
Satoko Iwasa, Hirotaka Ota, Kensuke Nishio, Mariko Ohtsu, Masafumi Kusunoki, Takahiro Gojoubori, Tetsuo Shirakawa, Masatake Asano
Toll-like receptors (TLR) recognize microbe-associated molecular patterns and induce the innate immune response. Among them, TLR5 recognizes the Gram-negative bacterial component flagellin. The aim of this study was to examine the expression of TLR5 in mouse salivary gland (SG). The SG was excised from 8- to 10-week-old female C57BL/6 mice. Salivary gland epithelial cells (SGECs) were purified and subjected to reverse transcription polymerase chain reaction (RT-PCR). Western blotting was performed to detect TLR5 expression at the protein level in several organs...
2016: Journal of Oral Science
Sandra Postel, Daniel Deredge, Daniel A Bonsor, Xiong Yu, Kay Diederichs, Saskia Helmsing, Aviv Vromen, Assaf Friedler, Michael Hust, Edward H Egelman, Dorothy Beckett, Patrick L Wintrode, Eric J Sundberg
Flagella are critical for bacterial motility and pathogenesis. The flagellar capping protein (FliD) regulates filament assembly by chaperoning and sorting flagellin (FliC) proteins after they traverse the hollow filament and exit the growing flagellum tip. In the absence of FliD, flagella are not formed resulting in impaired motility and infectivity. Here, we report the 2.2 Å resolution X-ray crystal structure of FliD from Pseudomonas aeruginosa, the first high-resolution structure of any FliD protein from any bacterium...
September 24, 2016: ELife
Malick Mbengue, Gildas Bourdais, Fabio Gervasi, Martina Beck, Ji Zhou, Thomas Spallek, Sebastian Bartels, Thomas Boller, Takashi Ueda, Hannah Kuhn, Silke Robatzek
Sensing of potential pathogenic bacteria is of critical importance for immunity. In plants, this involves plasma membrane-resident pattern recognition receptors, one of which is the FLAGELLIN SENSING 2 (FLS2) receptor kinase. Ligand-activated FLS2 receptors are internalized into endosomes. However, the extent to which these spatiotemporal dynamics are generally present among pattern recognition receptors (PRRs) and their regulation remain elusive. Using live-cell imaging, we show that at least three other receptor kinases associated with plant immunity, PEP RECEPTOR 1/2 (PEPR1/2) and EF-TU RECEPTOR (EFR), internalize in a ligand-specific manner...
September 27, 2016: Proceedings of the National Academy of Sciences of the United States of America
Caroline E Foust-Wright, Samantha J Pulliam, Rebecca Posthuma Batalden, Tucker K Berk, Milena M Weinstein, May M Wakamatsu, Mark Phillippe
AIM: The effect of hormone levels on the stimulation of Toll-like receptor 5 (TLR5) in the bladder is unknown. We aimed to study the effect of estradiol and progesterone on TLR5 expression and function in human bladder epithelial cells. METHODS: After growing to near confluence, T24 human urinary bladder (HUB) cells were incubated in hormone-free (HF) media for 72 hours. Human urinary bladder cells were then incubated in (1) HF media, (2) estradiol media, (3) progesterone media, or (4) media containing estradiol and progesterone at physiologic concentrations...
September 20, 2016: Reproductive Sciences
Fiorella Di Claudio, Cecilia Muglia, Paola Smaldini, María Lucía Orsini Delgado, Fernando Trejo, Raúl Grigera, Guillermo Docena
Intestinal epithelial cell culture is important for biological, functional and immunological studies. Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel and reproducible method to prolong the survival of mouse and human cells. Cells were isolated following a standard procedure, and cultured on ordered-cow's collagen membranes. A prolonged cell life span was achieved; cells covered the complete surface of bio-membranes and showed a classical enterocyte morphology with high expression of enzymes supporting the possibility of cryopreservation...
September 14, 2016: Journal of Cellular Physiology
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