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#28479111   2017/05/08 Save this To Up

Co-utilization of a TLR5 agonist and nano-formulation of HIV-1 vaccine candidate leads to increased vaccine immunogenicity and decreased immunogenic dose: A preliminary study.

Vaccines currently available for AIDS show poor efficiency, demonstrating the need for new strategies to increase their immunogenicity. In this study, the HIV-1P24-Nef peptide was used as a model vaccine, followed by utilization of a novel strategy to increase its immunogenicity. There is a growing interest in using TLR agonists for vaccine formulations. Such molecules bind to their receptors on immune cells, especially the cell surface of antigen presenting cells, thereby activating these cells and inflammatory responses. In the present study, FLiC (flagellin molecule sequence from Pseudomonas aeruginosa) was used as a TLR5 agonist. In addition, PLGA nanoparticles were used as a transmitter system to enhance vaccine efficiency and its effective transfer to immune systems. In light of this, the P24-Nef peptide was conjugated to FLiC through chemical reactions. The HIV-1P24-Nef/FLiC conjugate was constructed as a nano-vaccine using PLGA particles. Subsequently, mice were immunized intradermally three times with three-week intervals with HIV-p24-Nef/FLiC/PLGA, HIV-p24-Nef/PLGA, FLiC/PLGA, PLGA, and PBS in two doses (20 and 5μg). Three weeks after the last booster injection, cell proliferation was assessed using the Brdu/ELISA assay, and cytotoxicity was evaluated by CFSE and splenocyte cytokine secretion (IL-4 and IFN-γ); in addition, IgG1 and IgG2a antibody isotype titers were determined using a commercial ELISA kit. Our results showed that Co-utilization of TLR5 and nano-particles not only improves vaccine immunogenicity but also decreases the immunogenic dose of vaccine candidate required. We showed that the immune system was effectively stimulated via the nano-vaccination strategy using the TLR5 agonists. The effect of this strategy showed variations in different parameters of the immune system; in this regard, cellular immune responses had a higher stimulation level, compared with humoral immune responses.

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Rabbit Anti-Human Androge 17β-Acetoxy-2α-bromo-5 (5α,16β)-N-Acetyl-16-[2 (5α,16β)-N-Acetyl-16-ac 5α-N-Acetyl-2'H-androst- 5α-N-Acetyl-2'H-androst- 3-O-Acetyl 5,14-Androstad 3-O-Acetyl-17-O-tert-buty 3β-O-Acetyl-androsta-5,1 Androstadienone C19H26O C 5α-Androstan-3β-ol � ∆2-Androstene-1α,17β-

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#27163839   2016/08/02 Save this To Up

A Recently Established Murine Model of Nasal Polyps Demonstrates Activation of B Cells, as Occurs in Human Nasal Polyps.

Animal model systems are invaluable for examining human diseases. Our laboratory recently established a mouse model of nasal polyps (NPs) and investigated similarities and differences between this mouse model and human NPs. We especially focus on the hypothesis that B cell activation occurs during NP generation in the murine model. After induction of ovalbumin-induced allergic rhinosinusitis, 6% ovalbumin and Staphylococcus aureus enterotoxin B (10 ng) were instilled into the nasal cavity of mice three times per week for 8 weeks. The development of structures that somewhat resemble NPs (which we will refer to as NPs) was confirmed by hematoxylin and eosin staining. The mRNA and protein levels of various inflammatory cell markers and mediators were measured by real-time PCR in nasal tissue and by ELISA in nasal lavage fluid (NLF), respectively. Total Ig isotype levels in NLF were also quantitated using the Mouse Ig Isotyping Multiplex kit (EMD Millipore, Billerica, MA) on a Luminex 200 instrument (Life Technologies, Grand Island, NY). Similar to human NPs, there were significant increases in gene expression of inflammatory cell markers, such as CD19, CD138, CD11c, and mast cell protease-6 in nasal tissue samples of the NP group compared with those of the control group. In further investigations of B cell activation, mRNA expressions of B cell activating factor and a proliferation-inducing ligand were found to be significantly increased in mouse NP tissue. B cell-activating factor protein concentration and IgA and IgG1 levels in NLF were significantly higher in the NP group compared with the control group. In this study, the NP mouse model demonstrated enhanced B cell responses, which are reminiscent of B cell responses in human NPs.

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#26868748   2016/02/12 Save this To Up

Vascular Endothelial Growth Factor (VEGF) Concentration Is Underestimated by Enzyme-Linked Immunosorbent Assay in the Presence of Anti-VEGF Drugs.

Commercially available enzyme-linked immunosorbent assay (ELISA) kits are often used to monitor vascular endothelial growth factor (VEGF) levels in exudative age-related macular degeneration. To test their accuracy, this study performed measurements using the ELISA kits in the presence of anti-VEGF drugs.

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#26767257   2016/01/15 Save this To Up

[Study on immunologic function of thioredoxin glutathione reductase from Schistosoma japonicum].

To study the immunogenicity and the immuno-protection of thioredoxin glutathione reductase from Schistosomajaponicum (SjTGR) against schistosome infection in mice.

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#26459027   2015/10/27 Save this To Up

Preparation and preliminary application of monoclonal antibodies to the receptor binding region of Clostridium difficile toxin B.

A previous nationwide Chinese epidemiological study revealed through isolation of A‑B+ Clostridium difficile strains, which produce toxin B (TcdB), but not toxin A TcdA, that the strains are widespread and more frequent in east Asian countries,. The development of a process capable of detecting TcdB is required in microbiological laboratories in order to facilitate the control of the A‑B+ C. difficile strains, however, no diagnostic reagents have been developed to date. The aim of the present study was to prepare monoclonal antibodies (mAbs) targeting the receptor binding region of TcdB (CDB3), and to establish a double‑antibody sandwich enzyme-linked immunosorbent assay (ds‑ELISA), which can be used for the diagnosis of C. difficile infection. The recombinant protein, glutathione S transferase (GST)‑CDB3 was expressed and purified using an Escherichia coli system. BALB/c mice were immunized with GST‑CDB3 recombinant protein. A hybridoma technique was used for the production of anti‑CDB3 mAb. The hybridoma clones were then screened using indirect ELISA, and anti‑CDB3 mAb was produced in the ascites of the BALB/c mice. Isotyping of anti‑CDB3 mAb was performed using an SBA Clonotyping system/horseradish peroxidase (HRP) ELISA kit. Protein G affinity chromatography was used for purification of anti‑CDB3 mAbs, and the titer and specificity of the anti‑CDB3 mAbs were assessed using indirect ELISA and western blot analysis, respectively. The ds‑ELISA was established using HRP‑labeled anti‑CDB3 mAbd, which were used to detect TcdB clinically in diarrhea stools. A total of five stable hybridoma cell clones (1E7B, 1F8D3, 2F8A6, 3B6F1 and 4A4G2) producing anti‑CDB3 mAb were established. The results of the present study indicated that the immunoglobulin (Ig)G isotype was predominant, as 1E7B2 IgG1 (λ), 2F8A6 IgG2a (κ) and 4A4G2 IgG1 (κ). In addition, the highest titer of anti‑CDB3 mAb (2F8A6 and 4A4G2) was 1:51,200. Western blotting revealed that the 2F8A6 and 4A4G2 mAbs recognized the CDB3 protein specifically. Following anti‑CDB3 mAb (4A4G2) HRP‑labeling, the optimal working concentration was confirmed to be 1:400, and the concentration of coated antibody (2F8A6) was 20 µg/ml. The sensitivity of the ds‑ELISA was 73.33% for the A+B+ toxigenic C. difficile strains, and 86.67% for the A‑B+ toxigenic C. difficile strains, with a specificity of 100% for all. In conclusion, the present study successfully developed novel mAbs specific to CDB3, and developed a ds-ELISA kit with high specificity and sensitivity for the rapid detection of TcdB. This offers a useful tool for the diagnostic assessment of TcdB.

1143 related Products with: Preparation and preliminary application of monoclonal antibodies to the receptor binding region of Clostridium difficile toxin B.

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#26419907   2015/10/20 Save this To Up

Versatile and Amplified Biosensing through Enzymatic Cascade Reaction by Coupling Alkaline Phosphatase in Situ Generation of Photoresponsive Nanozyme.

The alkaline phosphatase (ALP) biocatalysis followed by the in situ enzymatic generation of a visible light responsive nanozyme is coupled to elucidate a novel amplification strategy by enzymatic cascade reaction for versatile biosensing. The enzymatic hydrolysis of o-phosphonoxyphenol (OPP) to catechol (CA) by ALP is allowed to coordinate on the surface of TiO2 nanoparticles (NPs) due to the specificity and high affinity of enediol ligands to Ti(IV). Upon the stimuli by CA generated from ALP, the inert TiO2 NPs is activated, which demonstrates highly efficient oxidase mimicking activity for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light (λ ≥ 400 nm) irradiation utilizing dissolved oxygen as an electron acceptor. On the basis of the cascade reaction of ALP and the nanozyme of CA coordinated TiO2 (TiO2-CA) NPs, we design exquisitely colorimetric biosensors for probing ALP activity and its inhibitor of 2, 4-dichlorophenoxyacetic acid (2,4-DA). Quantitative probing of ALP activity in a wide linear range from 0.01 to 150 U/L with the detection limit of 0.002 U/L is realized, which endows the methodology with sufficiently high sensitivity for potentially practical applications in real samples of human serum (ALP level of 40-190 U/L for adults). In addition, a novel immunoassay protocol by taking mouse IgG as an example is validated using the ALP/nanozyme cascade amplification reaction as the signal transducer. A low detection limit of 2.0 pg/mL is attained for mouse IgG, which is 4500-fold lower than that of the standard enzyme-linked immuno-sorbent assay (ELISA) kit. Although only mouse IgG is used as a proof-of-concept in our experiment, we believe that this approach is generalizable to be readily extended to other ELISA systems. This methodology opens a new horizon for amplified and versatile biosensing including probing ALP activity and following ALP-based ELISA immunoassays.

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#25993986   2015/05/21 Save this To Up

Development of an improved competitive ELISA based on a monoclonal antibody against lipopolysaccharide for the detection of bovine brucellosis.

Brucellosis is the most common bacterial zoonosis, and serological tests are routinely used in brucellosis control and eradication programs. In order to improve the accuracy of serological diagnostic method used in bovine brucellosis detection, this study developed an improved competitive ELISA with higher specificity and good sensitivity.

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#25926946   2015/04/30 Save this To Up

Production and Characterization of Monoclonal Antibodies against Human Prostate Specific Antigen.

Prostate Specific Antigen (PSA) is an important laboratory marker for diagnosis of prostatic cancer. Thus, development of diagnostic tools specific for PSA plays an important role in screening, monitoring and early diagnosis of prostate cancer. In this paper, the production and characterization of a panel of murine monoclonal antibodies (mAbs) against PSA have been presented.

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#25789225   2015/03/19 Save this To Up

Large scale generation and characterization of anti-human IgA monoclonal antibody in ascitic fluid of BALB/c mice.

Monoclonal antibodies are potentially powerful tools used in biomedical research, diagnosis, and treatment of infectious diseases and cancers. The monoclonal antibody against Human IgA can be used as a diagnostic application to detect infectious diseases. The aim of this study was to improve an appropriate protocol for large-scale production of mAbs against IgA.

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#25785047   2015/03/18 Save this To Up

IL-1RA gene-transfected bone marrow-derived mesenchymal stem cells in APA microcapsules could alleviate rheumatoid arthritis.

In order to investigate the encapsulation of interleukin 1 receptor antagonist (IL-RA) gene-modified mesenchymal stem cells (MSCs) in alginate-poly-L-lysine (APA) microcapsules for the persistent delivery of interleukin 1 receptor antagonist (IL-RA) to treat Rheumatoid arthritis (RA).

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