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Çiğdem Yılmaz, Aycan Apak, Erkan Özcengiz, Gülay Özcengiz
Whooping cough (pertussis) is a highly contagious respiratory infection caused by Bordetella pertussis. Although availability of effective pertussis vaccines seems to decrease the incidence of the disease, B. pertussis circulation in population has not been eliminated. Thus, finding new protein candidates with high immune protective capacities is necessary to enhance the efficacy of current acellular pertussis (Pa) vaccines. In this study, iron superoxide dismutase (FeSOD) gene (sodB) was cloned, expressed in Escherichia coli and recombinant FeSOD protein was purified...
October 20, 2016: Microbiology and Immunology
Christopher A Seid, Kathryn M Jones, Jeroen Pollet, Brian Keegan, Elissa Hudspeth, Molly Hammond, Junfei Wei, C Patrick McAtee, Leroy Versteeg, Amanda Gutierrez, Zhuyun Liu, Bin Zhan, Jonathan L Respress, Ulrich Strych, Maria Elena Bottazzi, Peter J Hotez
A therapeutic vaccine for human Chagas disease is under development by the Sabin Vaccine Institute Product Development Partnership. The aim of the vaccine is to significantly reduce the parasite burden of Trypanosoma cruzi in humans, either as a standalone product or in combination with conventional chemotherapy. Vaccination of mice with Tc24 formulated with monophosphoryl-lipid A (MPLA) adjuvant results in a Th1 skewed immune response with elevated IgG2a and IFNγ levels and a statistically significant decrease in parasitemia following T...
October 13, 2016: Human Vaccines & Immunotherapeutics
Jaxaira Maggi, Katina Schinnerling, Bárbara Pesce, Catharien M Hilkens, Diego Catalán, Juan C Aguillón
Tolerogenic dendritic cells (DCs) are a promising tool to control T cell-mediated autoimmunity. Here, we evaluate the ability of dexamethasone-modulated and monophosphoryl lipid A (MPLA)-activated DCs [MPLA-tolerogenic DCs (tDCs)] to exert immunomodulatory effects on naive and memory CD4(+) T cells in an antigen-specific manner. For this purpose, MPLA-tDCs were loaded with purified protein derivative (PPD) as antigen and co-cultured with autologous naive or memory CD4(+) T cells. Lymphocytes were re-challenged with autologous PPD-pulsed mature DCs (mDCs), evaluating proliferation and cytokine production by flow cytometry...
2016: Frontiers in Immunology
Hyun Jong Noh, Young-Woock Noh, Min Beom Heo, Eun-Ha Kim, Su-Jin Park, Young-Il Kim, Young Ki Choi, Yong Taik Lim
Seasonal emerging infectious diseases such as influenza A impose substantial risk and need new translational strategies to achieve active immunomodulation. Here, a novel injectable pathogen-mimicking hydrogel (iPMH) that can enhance both cellular and humoral immune responses is suggested. By the help of poly(γ-glutamic acid) that has abundant carboxylate groups and dispersion helper function, hydrophobic immunostimulatory 3-O-desacyl-4'-monophosphoryl lipid A (MPLA) molecules and viral antigens (PR8, W150) can be successfully combined as pathogen-mimicking adjuvants...
September 27, 2016: Small
Alison J Scott, Bryn Flinders, Joanna Cappell, Tao Liang, Rebecca S Pelc, Bao Tran, David P A Kilgour, Ron M A Heeren, David R Goodlett, Robert K Ernst
The discovery of novel pathogenic mechanisms engaged during bacterial infections requires the evolution of advanced techniques. Here, we evaluate the dual polarity matrix norharmane (NRM) to improve detection of bacterial lipid A (endotoxin), from host and vector tissues infected with Francisella novicida (Fn). We evaluated NRM for improved detection and characterization of a wide range of lipids in both positive and negative polarities, including lipid A and phospholipids across a range of matrix assisted laser desorption-ionization (MALDI)-coupled applications...
September 19, 2016: Pathogens and Disease
Taise Natali Landgraf, Fabrício Freitas Fernandes, Gabriela Peron, Ademilson Panunto-Castelo
The lack of antifungals with low toxicity and short-term therapy for patients with paracoccidioidomycosis (PCM) led us to evaluate adjuvants in immunotherapeutic intervention. We have previously shown complete Freund's adjuvant (CFA) to be therapeutic on experimental PCM. Owing to CFA toxicity, here we tested adjuvants approved for clinical use or in preclinical phase in experimental mouse PCM. Of all, only monophosporyl lipid A (MPLA) demonstrates a beneficial effect, by reducing the fungal burden and increasing the concentrations of IFN-γ and TNF-α, which are immunoprotective in PCM...
September 17, 2016: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
Michael Fichter, Keti Piradashvili, Anette Pietrzak-Nguyen, Leah Pretsch, Gabor Kuhn, Susanne Strand, Markus Knuf, Fred Zepp, Frederik R Wurm, Volker Mailänder, Katharina Landfester, Stephan Gehring
Targeting antigen combined with adjuvants to hepatic antigen-presenting cells (APCs) is essential for the induction of intrahepatic T cellular immunity controlling and resolving viral infections of the liver. Intravenous injection of antigen-loaded nanoparticles is a promising approach for the delivery of antigens to liver APCs. Accordingly, polymeric nanocapsules (NCs) synthesized exclusively of hepatitis C virus non-structural protein 5A (NS5A) and the adjuvant monophosphoryl lipid A (MPLA) adsorbed to the nanocapsule surface were developed...
November 2016: Biomaterials
Kantrol Kumar Sahu, Ravi Shankar Pandey
Hepatitis B is one of the leading liver diseases and remains a major global health problem. Currently available vaccines provide protection but often results in weaker/minimal mucosal immunity. Thus the present study is devoted to the development and in-vivo exploration of the colonically delivered biomimetic nanoparticles which capably enhance humoral as well as cellular immune response. In present work, Hepatitis B surface antigen (HBsAg) entrapped nanoparticles containing Monophosphoryl lipid A (MPLA) (HB+L-NP) were prepared by solvent evaporation method and characterized for particle size (~210nm), shape, zeta potential (-24mV±0...
October 2016: International Immunopharmacology
Anette Pietrzak-Nguyen, Keti Piradashvili, Michael Fichter, Leah Pretsch, Fred Zepp, Frederik R Wurm, Katharina Landfester, Stephan Gehring
Chronic hepatitis B virus (HBV) infection is the most prevalent serious liver infection in the world. A frequent route of infection represents mother-to-child transmission. Efficient control of HBV replication depends on antigen-specific cellular immune response mediated by dendritic cells (DCs). Aim of the present study was to evaluate optimized adjuvant combinations, efficiently maturing monocyte-derived neonatal and adult dendritic cells (moDCs). In addition, the potential of polymeric HBsAg-nanocapsules (HBsAg-NCs) was investigated regarding up-take by moDCs and the subsequent induction of specific T cell responses in a human co-culture model...
August 8, 2016: Nanomedicine: Nanotechnology, Biology, and Medicine
María Victoria Humbert, Miao-Chiu Hung, Renee Phillips, Charlene Akoto, Alison Hill, Wei-Ming Tan, John Edward Heckels, Myron Christodoulides
The cbf gene from Neisseria meningitidis strain MC58 encoding the putative Cell Binding Factor (CBF, NMB0345/NEIS1825) protein was cloned into the pRSETA system and a ~36-kDa recombinant (r)CBF protein expressed in Escherichia coli and purified by metal affinity chromatography. High titres of rCBF antibodies were induced in mice following immunization with rCBF-saline, rCBF-Al(OH)3, rCBF-Liposomes or rCBF-Zwittergent (Zw) 3-14 micelles, both with and without incorporated monophosphoryl lipid A (MPLA) adjuvant...
2016: PloS One
Ilda Patrícia Ribeiro, Francisco Caramelo, Francisco Marques, Ana Domingues, Margarida Mesquita, Leonor Barroso, Hugo Prazeres, Maria José Julião, Isabel Poiares Baptista, Artur Ferreira, Joana Barbosa Melo, Isabel Marques Carreira
PURPOSE: Oral squamous cell carcinoma (OSCC) is a frequently occurring aggressive malignancy with a heterogeneous clinical behavior. Based on the paucity of specific early diagnostic and prognostic biomarkers, which hampers the appropriate treatment and, ultimately the development of novel targeted therapies, we aimed at identifying such biomarkers through a genetic and epigenetic analysis of these tumors. METHODS: 93 primary OSCCs were subjected to DNA copy number alteration (CNA) and methylation status analyses using methylation-specific multiplex ligation-dependent probe amplification (MS-MPLA)...
August 4, 2016: Cellular Oncology (Dordrecht)
Wagner Quintilio, Fábio Alessandro de Freitas, Dunia Rodriguez, Flavia Saldanha Kubrusly, Dimitri Yourtov, Cosue Miyaki, Luciana Cezar de Cerqueira Leite, Isaias Raw
A number of adjuvant formulations were assayed in mice immunized with 3.75 µg of A/California/7/2009 (H1N1) pdm09 influenza vaccine with vitamins A, D and/or E in emulsions or B2 and/or B9 combined with Bordetella pertussis MPLA and/or alum as adjuvants. Squalene was used as positive control, as well as MPLA with alum. The immune response was evaluated by a panel of tests, including a hemagglutination inhibition (HAI) test, ELISA for IgG, IgG1, and IgG2a and IFN-γ, IL-2, IL-6 and IL-10 quantification in splenocyte culture supernatant after stimulus with influenza antigen...
October 2016: Archives of Virology
James E Voss, Matthew S Macauley, Kenneth A Rogers, Francois Villinger, Lijie Duan, Liang Shang, Elizabeth A Fink, Raiees Andrabi, Arnaud D Colantonio, James E Robinson, R Paul Johnson, Dennis R Burton, Ashley T Haase
Vaccination with SIVmac239Δnef provides robust protection against subsequent challenge with wild type SIV, but safety issues have precluded designing an HIV-1 vaccine based on a live attenuated virus concept. Safe immunogens and adjuvants that could reproduce identified immune correlates of SIVmac239Δnef protection therefore offer an alternative path for development of an HIV vaccine. Here we describe SIV envelope trimeric gp41 (gp41t) immunogens based on a protective correlate of antibodies to gp41t concentrated on the path of virus entry by the neonatal Fc receptor (FcRn) in cervical vaginal epithelium...
July 13, 2016: AIDS
Xuanye Wu, Nils Schneider, Alina Platen, Indranil Mitra, Matthias Blazek, Roland Zengerle, Roland Schüle, Matthias Meier
Mammalian target of rapamycin (mTOR) is a central kinase integrating nutrient, energy, and metabolite signals. The kinase forms two distinct complexes: mTORC1 and mTORC2. mTORC1 plays an essential but undefined regulatory function for regeneration of adipose tissue. Analysis of mTOR in general is hampered by the complexity of regulatory mechanisms, including protein interactions and/or phosphorylation, in an ever-changing cellular microenvironment. Here, we developed a microfluidic large-scale integration chip platform for culturing and differentiating human adipose-derived stem cells (hASCs) in 128 separated microchambers under standardized nutrient conditions over 3 wk...
July 19, 2016: Proceedings of the National Academy of Sciences of the United States of America
Hayk Davtyan, Karen Zagorski, Harinda Rajapaksha, Armine Hovakimyan, Arpine Davtyan, Irina Petrushina, Konstantin Kazarian, David H Cribbs, Nikolai Petrovsky, Michael G Agadjanyan, Anahit Ghochikyan
Although β-amyloid (Aβ) may be the primary driver of Alzheimer's disease (AD) pathology, accumulation of pathological tau correlates with dementia in AD patients. Thus, the prevention/inhibition of AD may require vaccine/s targeting Aβ and tau simultaneously or sequentially. Since high antibody titers are required for AD vaccine efficacy, we have decided to generate vaccines, targeting Aβ (AV-1959R), Tau (AV-1980R) or Aβ/tau (AV-1953R) B cell epitopes, based on immunogenic MultiTEP platform and evaluate the immunogenicity of these vaccines formulated with Advax(CpG), delta inulin, Alhydrogel(®), Montanide-ISA51, Montanide-ISA720, MPLA-SM pharmaceutical grade adjuvants...
2016: Scientific Reports
Antonio Hernandez, Julia K Bohannon, Liming Luan, Benjamin A Fensterheim, Yin Guo, Naeem K Patil, Chase McAdams, Jingbin Wang, Edward R Sherwood
Treatment with the TLR4 agonist MPLA augments innate resistance to common bacterial pathogens. However, the cellular and molecular mechanisms by which MPLA augments innate immunocyte functions are not well characterized. This study examined the importance of MyD88- and TRIF-dependent signaling for leukocyte mobilization, recruitment, and activation following administration of MPLA. MPLA potently induced MyD88- and TRIF-dependent signaling. A single injection of MPLA caused rapid mobilization and recruitment of neutrophils, a response that was largely mediated by the chemokines CXCL1 and -2 and the hemopoietic factor G-CSF...
June 27, 2016: Journal of Leukocyte Biology
Stefan Schülke, Lothar Vogel, Ann-Christine Junker, Kay-Martin Hanschmann, Adam Flaczyk, Stefan Vieths, Stephan Scheurer
Background. The detoxified TLR4-ligand Monophosphoryl Lipid A (MPLA) is the first approved TLR-agonist used as adjuvant in licensed vaccines but has not yet been explored as part of conjugated vaccines. Objective. To investigate the immune-modulating properties of a fusion protein consisting of MPLA and Ovalbumin (MPLA : Ova). Results. MPLA and Ova were chemically coupled by stable carbamate linkage. MPLA : Ova was highly pure without detectable product-related impurities by either noncoupled MPLA or Ova...
2016: Journal of Immunology Research
Wellcy Gonçalves Teixeira, Fabiana Kalina Marques, Maíra Cristina Menezes Freire
OBJECTIVE: To describe the chromosomal alterations in patients with mental retardation (MR) using G-banding karyotype analysis. METHOD: A retrospective study of the results G-banding karyotype analysis of 369 patients investigated for MR was performed. Based on the structural rearrangements found, the authors searched all chromosomal regions related with breakpoints, and these were compared with the literature on MR and databases. RESULTS: 338 (91...
May 2016: Revista da Associação Médica Brasileira
Maritsa Margaroni, Maria Agallou, Katerina Kontonikola, Konstantina Karidi, Olga Kammona, Costas Kiparissides, Catherine Gaitanaki, Evdokia Karagouni
Poly(lactide-co-glycolide) nanoparticles (PLGA NPs) represent a new approach for vaccine delivery due to their ability to be taken up by phagocytes and to activate immune responses. In the present study PLGA NPs were surface-modified with a TNFα mimicking peptide, and encapsulated soluble Leishmania antigens (sLiAg) and MPLA adjuvant. The synthesized PLGA NPs exhibited low cytotoxicity levels, while surface-modified NPs were more efficiently taken up by dendritic cells (DCs). The prepared nanoformulations induced maturation and functional differentiation of DCs by elevating co-stimulatory molecule levels and stimulating IL-12 and IL-10 production...
August 2016: European Journal of Pharmaceutics and Biopharmaceutics
Devendra K Rai, Fayna Diaz-San Segundo, Elizabeth Schafer, Thomas G Burrage, Luis L Rodriguez, Teresa de Los Santos, Paul D Hoeprich, Elizabeth Rieder
Here, we engineered two FMD viruses with histidine residues inserted into or fused to the FMDV capsid. Both 6xHis viruses exhibited growth kinetics, plaque morphologies and antigenic characteristics similar to wild-type virus. The 6xHis tag allowed one-step purification of the mutant virions by Co(2+) affinity columns. Electron microscopy and biochemical assays showed that the 6xHis FMDVs readily assembled into antigen: adjuvant complexes in solution, by conjugating with Ni(2+)-chelated nanolipoprotein and monophosphoryl lipid A adjuvant (MPLA:NiNLP)...
August 2016: Virology
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