Chitosan polysaccharide suppress toll like receptor dependent immune response [Çitosan polisakkaridi toll benzeri reseptöre bağlı bağışıklık yanıtını baskılar]

Objectives: Chitosan is a widely used vaccine or anti-cancer delivery vehicle. In this study, we investigated the immunomodulatory effect of chitosan/pIC nanocomplexes on mouse immune cells. Materials and methods: Proliferative and cytotoxic features of chitosan were tested via CCK-8 assay on RAW 264. 7. IL-1β production was assessed via ELISA from PEC supernatants. TNF-α, and NO induction from chitosan treated RAW cells detected by ELISA and Griess assay, respectively. mRNA message levels of TLRs and cytokines on macrophages in response to chitosan/pIC nanocomplex treatments were evaluated by RT-PCR. Results: Results revealed that chitosan is non-toxic to cells, however, proliferative capacities of macrophages were reduced by chitosan administration. Mouse PECs treated with chitosan, led to NLRP3 dependent inflammasome activation as evidenced by dose-dependent IL-1β secretion. Chitosan/pIC nanocomplexes did not improve immunostimulatory action of pIC on RAW cells, since TNF-α and NO productions remained unaltered. Expression levels of several TLRs, CXCL-16 and IFN-α messages from mouse splenocytes were down regulated in response to chitosan/pIC nanocomplex treatment. Conclusion: Our results revealed that chitosan is an anti-proliferative and inflammasome triggering macromolecule on immune cells. Utilization of chitosan as a carrier system is of concern for immunotherapeutic applications. © 2015 Turkish Journal of Immunology

Immunostimulatory activity of polysaccharide-poly(I:C) nanoparticles

Cataloged from PDF version of article.Immunostimulatory properties of mushroom derived polysaccharides (PS) as stand-alone agents were tested. Next. PS were nanocomplexed with polyI:C (pIC) to yield stable nanoparticles around 200 nm in size evidenced by atomic force microscopy and dynamic light scattering analyses. PSs were selectively engaged by cells expressing TLR2 and initiated NF kappa B dependent signaling cascade leading to a Th1-biased cytokine/chemokine secretion in addition to bactericidal nitric oxide (NO) production from macrophages. Moreover, cells treated with nanoparticles led to synergistic IL6, production and upregulation of TNF alpha, MIP3 alpha, IFN gamma and IP10 transcript expression. In mice, PS-Ovalbumin-pIC formulation surpassed anti-OVA IgG responses when compared to either PS-OVA or pIC-OVA mediated immunity. Our results revealed that signal transduction initiated both by TLR2 and TLR3 via co-delivery of pIC by PS in nanoparticle depot delivery system is an effective immunization strategy. The present work implicate that the PS and nucleic acid based nanoparticle approach along with protein antigens can be harnessed to prevent infectious diseases. (C) 2011 Elsevier Ltd. All rights reserve

Encapsulation of two different TLR ligands into liposomes confer protective immunity and prevent tumor development

Nucleic acid-based Toll-like receptor (TLR) ligands are promising adjuvants and immunotherapeutic agents. Combination of TLR ligands potentiates immune response by providing synergistic immune activity via triggering different signaling pathways and may impact antigen dependent T-cell immune memory. However, their short circulation time due to nuclease attack hampers their clinical performance. Liposomes offer inclusion of protein and nucleic acid-based drugs with high encapsulation efficiency and drug loading. Furthermore, they protect cargo from enzymatic cleavage while providing stability, and enhancing biological activity. Herein, we aimed to develop a liposomal carrier system co-encapsulating TLR3 (polyinosinic-polycytidylic acid; poly(I:C)) and TLR9 (oligodeoxynucleotides (ODN) expressing unmethylated CpG motifs; CpG ODN) ligands as immunoadjuvants together with protein antigen. To demonstrate that this depot system not only induce synergistic innate immune activation but also boost antigen-dependent immune response, we analyzed the potency of dual ligand encapsulated liposomes in long-term cancer protection assay. Data revealed that CpG ODN and poly(I:C) co-encapsulation significantly enhanced cytokine production from spleen cells. Activation and maturation of dendritic cells as well as bactericidal potency of macrophages along with internalization capacity of ligands were elevated upon incubation with liposomes co-encapsulating CpG ODN and poly(I:C). Immunization with co-encapsulated liposomes induced OVA-specific Th1-biased immunity which persisted for eight months post-booster injection. Subsequent challenge with OVA-expressing tumor cell line, E.G7, demonstrated that mice immunized with liposomes co-encapsulating dual ligands had significantly slower tumor progression. Tumor clearance was dependent on OVA-specific cytotoxic memory T-cells. These results suggest that liposomes co-encapsulating TLR3 and TLR9 ligands and a specific cancer antigen could be developed as a preventive cancer vaccine. � 2017 Elsevier B.V

The effects of an insertion in the 5 ' UTR of the AMCase on gene expression and pulmonary functions

Cataloged from PDF version of article.Background: Studies regarding the physiological role of acidic mammalian chitinase (AMCase) and the effects of its genetic variants on asthma have produced conflicting results. Objectives: We aimed to determine the genetic variants in the AMCase gene that could regulate the gene expression and thus influence disease severity. Methods: Genetic variants of the AMCase gene were determined by sequencing of asthmatics and healthy controls in up to -1 kb in the promoter region and exon 1 and 2. In an association study, a population of asthmatic (n = 504) and healthy Turkish children (n = 188) were genotyped for the observed SNPs. A replication study was performed in a North American adult population of patients with mild (n = 317) and severe (n = 145) asthma. The functional properties of the insertion were determined by promoter reporter assay, electromobility shift assay and transcription factor ELISA experiments. Results: Of the identified SNPs, only a ten base pair insertion (CAATCTAGGC) in the 5'UTR region of exon 2 was significantly associated with lower FEV(1) (beta = -14.63 SE = 6.241, P = 0.019) in Turkish children with asthma. However, in the adult population, the same insertion showed a trend toward higher FEV(1). The insertion was shown to have enhancer activity and the mutant probe possessing the insertion had higher binding affinity for the nuclear extracts. Conclusion: Our study shows that a ten base pair insertion in the 5'UTR region of AMCase gene may modify gene expression and thus may affect the severity of asthma. However, its effects appear to be different in different populations. (C) 2011 Elsevier Ltd. All rights reserved

PREPARATION AND CHARACTERIZATION OF PERLITE-FILLED HIGH-DENSITY POLYETHYLENES .2. THERMAL AND FLOW PROPERTIES

Thermal and flow properties of perlite-filled high-density polyethylenes (HDPE), studied in the first part of the paper, are discussed. Maximum peak temperatures of endotherms (T-mp) and the corresponding relative peak heights were examined by differential scanning calorimetry (DSC). The data obtained from a Brabender torque rheometer were evaluated to find the melt viscosities at low shear rate during preparation of perlite-HDPE composites. The filler concentration, types of HDPEs, and the use of gamma-APS are found to be effective and influential factors on the thermal and flow properties of these composites. (C) 1994 John Wiley and Sons, Inc

PREPARATION AND CHARACTERIZATION OF PERLITE-FILLED HIGH-DENSITY POLYETHYLENES .1. MECHANICAL-PROPERTIES

Four different types of high-density polyethylenes (HDPE) were blended with perlite at different concentrations. Silane coupling agent gamma-aminopropyltriethoxy silane (gamma-APS, A-1100) was used to enhance the adhesion between perlite and HDPEs. Ultimate tensile strength and elastic modulus increased as the perlite content increased, while ultimate elongations decreased with the increasing amount of perlite. Exceptional variations in the measured properties are explained in terms of the differences in polyethylenes used in the composites. The biggest improvement in ultimate tensile strength was observed in the highly crystalline polyethylene; on the other hand, in the absence of silane coupling agent the high molecular weight polyethylene showed the least improvement in tensile strength. The effect of branching in HDPE composites was demonstrated. The enhancement of interfacial adhesion by using a coupling agent was also examined by scanning electron microscopy (SEM). (C) 1994 John Wiley and Sons, Inc

Effect of five woven fiber reinforcements on the impact and transverse strength of a denture base resin

Statement of problem. Fracture strength of denture base resins is of great concern, and many approaches have been used to strengthen acrylic resin dentures