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- Combined meningococcal serogroup A and W135 outer-membrane vesicles activate cell-mediated immunity and long-term memory responses against non-covalent capsular polysaccharide A. [JOURNAL ARTICLE]
- Immunol Res 2013 May 10.
Outer-membrane vesicles (OMVs) have inherent adjuvant properties, and many vaccines use OMV as vaccine components. Utilizing the adjuvant properties of OMV could lead to the formulation of vaccines that are less expensive and potentially more immunogenic than covalently conjugated polysaccharide vaccines. We evaluated the adjuvant effect in Balb/c mice of combinations of OMV from Neisseria meningitidis serogroup A and W135 as compared to that of the non-covalently conjugated capsular polysaccharide A. Both antigens were adsorbed onto aluminum hydroxide. The mice were given a booster dose of plain polysaccharide A to stimulate an immunologic memory response. Subclasses determination and cytokine assays demonstrated the capacity of OMV to induce a IgG2a/IgG2b isotype profile and IFN-γ production, suggesting the induction of a Th1 pattern immune response. Lymphoproliferative responses to OMVs were high, with affinity maturation of antibodies observed. Bactericidal titers after the booster dose were also observed. Memory B cells and long-term memory T cells were also detected. The results of this study indicate that combined meningococcal serogroup A and W135 OMV can activate cell-mediated immunity and induce a long-term memory response.
- Immune responses to tyrophagus putrescentiae-induced airway inflammation in mice. [Journal Article, Research Support, Non-U.S. Gov't]
- J Investig Allergol Clin Immunol 2013; 23(1):20-9.
Storage mites are a source of aeroallergens that affect patients with allergic rhinitis and asthma. Tyrophagus putrescentiae is a causative factor of airway hypersensitivity, but the mechanisms and pathogenesis of Tputrescentiae-induced allergy are not well understood.This study aimed to develop a murine model of T putrescentiae-induced allergic asthma.Immune responses and physiologic variations in immunoglobulins (Ig), leukocyte subpopulations, cytokines, gene expression, pulmonary function, and lung pathology were evaluated after intraperitoneal sensitization and intratracheal challenge with crude extract of T putrescentiae.After sensitization with aluminum hydroxide and challenge with T putrescentiae in mice, levels of T putrescentiae-specific IgE and IgG1 in sera increased significantly compared to the normal saline group (P < .01): Values for inflammatory leukocytes (neutrophils and eosinophils) and cytokines (interleukin [IL] 4, IL-5, and IL-13) increased significantly after sensitization. In terms of pulmonary function, pause values were significantly enhanced in T putrescentiae-sensitized mice after intratracheal challenge with T putrescentiae (P < .05). Expression of type 2 helper T cell (T(H)2)-related genes (IL4, IL5, IL13, and RANTES), T(H)2-specific transcription factor (GATA-3), and proinflammatory genes (IL6), and T(H)(H)17-related genes (IL17F) increased significantly after airway challenge. Sensitization with T putrescentiae crude extract led to inflammation of lung tissue, thickening of the tracheal wall, and tracheal rupture.Intraperitoneal sensitization followed by intratracheal challenge with crude extract of T putrescentiae can induce airway inflammation in BALB/c mice. The symptoms observed in a mouse model of allergic asthma, in terms of immune and clinical parameters, are reminiscent of the symptoms of allergic asthma in humans. A mouse model can be used to evaluate the therapeutic effectiveness of drugs on T putrescentiae-induced airway inflammation in humans.
- Pore-size dependent immunogenic activity of mesoporous silica-based adjuvants in cancer immunotherapy. [JOURNAL ARTICLE]
- J Biomed Mater Res A 2013 May 7.
Commonly used aluminum hydroxide (Alum) adjuvant provokes a strong type 2 helper T cell (Th2) response for mediating antibody production, but is rather ineffective for disease prevention that requires type 1 helper T cell (Th1) response for mediating cellular immunity in human vaccination. Here, for the purpose of inducing Th1 anti-tumor immunity, a mesoporous silica (MS)-based adjuvant is prepared. Three kinds of MS particles with nearly identical particle size and surface area but different pore sizes of 4, 7 and 10 nm were prepared. No serious in vitro cytotoxicity was observed for the MS particles at 5, 20, 50 and 100 μg/mL. Pathogen-associated molecular patterns (PAMPs) were immobilized with apatite (Ap) on MS to prepare the MS-based and PAMP-loaded adjuvants (MS-Ap-PAMP adjuvants). Macrophage-like cells cultured in the presence of MS-Ap-PAMP adjuvant with a MS pore size of 10 nm showed the maximum in vitro immunogenic activity. Injection of the MS-Ap-PAMP adjuvant with a MS pore size of 10 nm in combination with liquid-nitrogen-treated tumor tissue (derived from Lewis lung carcinoma cells) to C57BL/6 mice markedly inhibited the development of re-challenged tumor in vivo, while no such antitumor immunity was induced in injection of Alum mixed with PAMP in combination with liquid-nitrogen-treated tumor tissue. The MS-Ap-PAMP adjuvant contributed to the elicitation of a potent systemic Th1 antitumor immunity in vivo.
- The simultaneous removal of calcium, magnesium and chloride ions from industrial wastewater using magnesium-aluminum oxide. [Journal Article]
- J Environ Sci Health A Tox Hazard Subst Environ Eng 2013; 48(10):1225-30.
In this article, a method for simultaneous removal of calcium, magnesium and chloride by using Mg0.80Al0.20O1.10 as a Magnesium-Aluminum oxide (Mg‒Al oxide) was investigated. Mg‒Al oxide obtained by thermal decomposition of the Mg-Al layered double hydroxide (Mg-Al LDH). The synthesized Mg‒Al oxide were characterized with respect to nitrogen physicosorption, X-ray diffraction (XRD) and field emission scan electron microscopy (FESEM) morphology. Due to high anion-exchange capacity of Mg‒Al oxide, it was employed in simultaneously removal of Cl(-), Mg(+2) and Ca(+2) from distiller waste of a sodium carbonate production factory. For this purpose, experiments were designed to evaluate the effects of quantity of Mg‒Al oxide, temperature and time on the removal process. The removal of Cl(-), Mg(+2) and Ca(+2) from wastewater was found 93.9%, 93.74% and 93.25% at 60°C after 0.5 h, respectively. Results showed that the removal of Cl(-), Mg(+2) and Ca(+2) by Mg‒Al oxide increased with increasing temperature, time and Mg‒Al oxide quantity.
- Experimental etch-and-rinse adhesives doped with bioactive calcium silicate-based micro-fillers to generate therapeutic resin-dentin interfaces. [JOURNAL ARTICLE]
- Dent Mater 2013 Apr 29.
OBJECTIVES:This study aimed at evaluating the therapeutic bioactive effects on the bond strength of three experimental bonding agents containing modified Portland cement-based micro-fillers applied to acid-etched dentin and submitted to aging in simulated body fluid solution (SBS). Confocal laser (CLSM) and scanning electron microscopy (SEM) were also performed.
METHODS:A type-I ordinary Portland cement was tailored using different compounds such as sodium-calcium-aluminum-magnesium silicate hydroxide (HOPC), aluminum-magnesium-carbonate hydroxide hydrates (HCPMM) and titanium oxide (HPCTO) to create three bioactive micro-fillers. A resin blend mainly constituted by Bis-GMA, PMDM and HEMA was used as control (RES-Ctr) or mixed with each micro-filler to create three experimental bonding agents: (i) Res-HOPC, (ii) Res-HCPMM and (iii) Res-HPCTO. The bonding agents were applied onto 37% H3PO4-etched dentin and light-cured for 30s. After build-ups, they were prepared for micro-tensile bond strength (μTBS) and tested after 24h or 6 months of SBS storage. SEM analysis was performed after de-bonding, while CLSM was used to evaluate the ultra-morphology/nanoleakage and the mineral deposition at the resin-dentin interface.
RESULTS:High μTBS values were achieved in all groups after 24h. Only Res-HOPC and Res-HCPMM showed stable μTBS after SBS storage (6 months). All the resin-dentin interfaces created using the bonding agents containing the bioactive micro-fillers tested in this study showed an evident reduction of nanoleakage and mineral deposition after SBS storage.
CONCLUSION:Resin bonding systems containing specifically tailored Portland cement micro-fillers may promote a therapeutic mineral deposition within the hybrid layer and increase the durability of the resin-dentin bond.
- TRPM2 channels are not required for acute airway inflammation in OVA-induced severe allergic asthma in mice. [Journal Article]
- J Inflamm (Lond) 2013; 10(1):19.
Airway inflammation and asthma have been linked to oxidative stress and the melastatin-related transient receptor potential cation channel, member 2 (TRPM2), which can be activated by reactive oxygen species (ROS), has emerged as a potential therapeutic target for inflammatory diseases.Using TRPM2 deficient (TRPM2-/-) mice, we investigated whether the TRPM2 ion channel, which mediates calcium (Ca2+) influx and lysosomal Ca2+ release, plays a role in the pathophysiology of severe allergic asthma in mouse.Severe allergic asthma was initiated in wild type (WT) and TRPM2-/- mice by repeated sensitization with ovalbumin (OVA)/aluminum hydroxide on Days 0, 7 and 14, followed by intranasal challenge on Days 21, 22 and 23. Mice were investigated for the presence of airway responsiveness, airway inflammation, production of allergen-specific antibodies, cytokine response and lung pathology.The absence of TRPM2 channels has no obvious effect on major etiologic markers of severe allergic asthma in this mouse model. Neither airway resistance nor mucus production are affected in TRPM2-/- mice. TRPM2 channel ablation also does not alter airway inflammation or immunocyte infiltration and does not affect antibody response or cytokine levels.TRPM2 is not required for airway inflammation in OVA-induced severe allergic asthma in mice. Accordingly, TRPM2 might not be a suitable therapeutic target for airway inflammation caused by allergens in humans.
- Involvement of NFκB in the Production of Chemokines by Rat and Human Conjunctival Cells Cultured Under Allergenic Conditions. [JOURNAL ARTICLE]
- Curr Eye Res 2013 Apr 26.
Purpose:The purpose of present studies was to determine the involvement of NFκB and STAT6 transcription factors in the production of cytokines by the fibroblasts and epithelial cells in conjunctiva.
Methods:An in vitro model of allergic conjunctivitis was developed by sensitizing and challenging rat mast cells with anti-dinitrophenyl (DNP) IgE and DNP-BSA, and then using the conditioned medium to stimulate rat conjunctival fibroblasts. Chemokines (eotaxin-1, IL-8, and RANTES -- Regulated and Normal T cell Expressed and Secreted) released from cells into the medium was determined by ELISA. Human conjunctival fibroblasts and epithelial cells were also directly stimulated with exogenous cytokines tumor necrosis factor (TNF)-α or IL-4. Degradation of IκB-α and phosphorylation of STAT6 were assessed by immunoblotting. For inhibition of NFκB or STAT6 activation, upstream regulators IκB kinase and Janus protein tyrosine kinases (JAK) were inhibited by use of BMS-345541 and JAK inhibitor 1. An in vivo model of conjunctivitis was also produced in rats by intraperitoneal injection of ovalbumin (OA) with aluminum hydroxide and challenge at 21 d with OA eye drops.
Results:Stimulated rat mast cells released TNF-α and IL-4. TNF-α induced NFκB activation in rat and human conjunctival fibroblasts and epithelial cells, and caused production and release of cytokines IL-8 and RANTES. IL-4 activation of STAT6 did not cause release of these cytokines. Only fibroblasts produced the eosinophil-recruiting cytokine, eotaxin-1, after treatment with TNF-α- plus IL-4. As observed in the cultured cells, allergic stimulation in the in vivo model caused degradation of IκB-α in conjunctiva, and infiltration of eosinophils and other inflammatory cells.
Conclusion:Activated NFκB was found to be a major transcription factor for the release of cytokines from conjunctival cells and intensification of the allergic response. Inhibition of the NFκB pathway by therapeutic drugs may be an important objective for the treatment of human allergic conjunctivitis.
- Determination of N-nitrosodiethanolamine in cosmetic products by headspace solid phase microextraction using a novel aluminum hydroxide grafted fused silica fiber followed by gas chromatography-mass spectrometry analysis. [Journal Article]
- Talanta 2013 Feb 15.:347-53.
A method based on headspace solid phase microextraction with a new fiber, coupled with gas chromatography-mass spectrometry was developed for the determination of NDELA in cosmetic samples. The fiber provides Lewis acid-base interaction between its surface and analyte functional groups. The fiber was prepared by grafting aluminum tri-tert-butoxide on the surface of a fused silica. The optimization of SPME conditions showed that NDELA can be most effectively extracted at 70°C, in 15min, with a sample volume of 0.5 (Vs/Vt), stirring rate of 150rpm, desorption time of 5min, desorption temperature of 260°C and at 12.5% (w/w) concentration of NaCl. Under the optimized conditions, LOD of 1μgKg(-1) and a calibration curve with correlation coefficients greater than 0.9897 and a linearity range from 6 to 10000μgKg(-1) were obtained. The intra-day and inter-day precision and accuracy were evaluated at four concentration levels. All of the values for accuracy and precision were lower than the acceptable limit of 15%. The fiber to fiber repeatability was 8.7%. The method was applied for the analysis of real samples including hair shampoo, body shampoo, dishwashing liquid and hand washing liquid. Relative recoveries were achieved in the range of 95-99%.
- Dietary L-proline supplementation confers immunostimulatory effects on inactivated Pasteurella multocida vaccine immunized mice. [JOURNAL ARTICLE]
- Amino Acids 2013 Apr 13.
This study was conducted to determine the immunostimulatory effect of L-proline on inactivated vaccine immunized mice. Ninety-five female KM mice were randomly divided into five groups: (1) mice received dietary supplementation with 0.4 % L-proline and immunized with inactivated vaccine (V-P group); (2) mice received dietary supplementation with 0.3 % L-alanine (isonitrogenous control) and immunized with inactivated vaccine (V-A group, negative control); (3) mice were immunized with inactivated vaccine with oil adjuvant (V-O group, positive control); (4) mice were immunized with inactivated vaccine with aluminum hydroxide adjuvant (V-H group, positive control); (5) mice immunized with phosphate-buffered saline (control group). All mice were dead in the control group between 36 and 48 h post infection. Mice in the V-P group showed 100 % protection after challenge with P. multocida serotype A (CQ2) at dose of 4.4 × 10(5) CFU (2LD50). Meanwhile, serum antibody titers in the V-P group were higher than those in the V-A group before infection and those in the V-A and V-O groups at 36 h post infection. Moreover, serum IL-1β levels in the V-P group were lower than those in V-O group. Furthermore, serum GSH-PX levels in the V-P group were higher than those in the V-A and V-O groups. Collectively, dietary proline supplementation confers beneficial immunostimulatory effects in inactivated P. multocida vaccine immunized mice.
- Stabilization of a recombinant ricin toxin A subunit vaccine through lyophilization. [JOURNAL ARTICLE]
- Eur J Pharm Biopharm 2013 Apr 10.
Lyophilization was used to prepare dry, glassy solid vaccine formulations of recombinant ricin toxin A-chain containing suspensions of colloidal aluminum hydroxide adjuvant. Four lyophilized formulations were prepared by using combinations of rapid or slow cooling during lyophilization and one of two buffers, histidine or ammonium acetate. Trehalose was used as the stabilizing excipient. Aggregation of the colloidal aluminum hydroxide suspension was reduced in formulations processed with a rapid cooling rate. Aluminum hydroxide particle size distributions, glass transition temperatures, water contents, and immunogenicities of lyophilized vaccines were independent of incubation time at 40°C for up to 15weeks. Mice immunized with reconstituted ricin toxin subunit A (RTA) vaccines produced RTA-specific antibodies and toxin-neutralizing antibodies (TNAs) regardless of the length of high temperature vaccine storage or the degree of aluminum adjuvant aggregation that occurred during lyophilization. In murine studies, lyophilized formulations of vaccines conferred protection against exposure to lethal doses of ricin, even after the lyophilized formulations had been stored at 40°C for 4weeks. A corresponding liquid formulation of vaccine stored at 40°C elicited RTA-specific antibody titers but failed to confer immunity during a ricin challenge.