ovalbumin and hydroxide-ion

Quercetin (Que), kaempferol (Kaem), isorhamnetin (Irh), and myricetin (Myri) are typical flavonols that are abundant in plant resources. This research investigated their ability in attenuating harmful glycation product formation and the effect of hydroxyl substitution. The inhibition mechanisms were elucidated by fluorescence spectroscopy and nano-liquid chromatography Orbitrap tandem mass spectrometry. The results indicated that the 3'-OH on the B-ring is critical in alleviating harmful glycation product formation, methylation reduced its inhibition, and the 5'-OH showed much less contribution than the 3'-OH. Que showed the strongest suppression on initial product, 5-hydroxymethylfurfural, and advanced glycation end product formation, with the corresponding percentage inhibitions at 36.58 μM of 81.1, 56.9, and 95.4%. Que and Myri also clearly inhibited fructosamine and acrylaminde production, while no suppression was observed by Irh and Kaem. The number of glycated sites was reduced from ten to seven, five, six, and nine, respectively, when 36.58 μM Que, Myri, Kaem, and Irh was added. Suppressing the conformational changes of ovalbumin induced by glycation, trapping dicarbonyl compounds, altering the microenvironment around tryptophan, and reducing the glycation activity of potential sites were the major inhibition mechanisms. These results suggest that Que and Myri may be promising natural agents for inhibiting harmful glycation and provide theoretical support for the effective screening of natural antiglycation reagents.

Topics: Animals; Cattle; Flavonols; Glycation End Products, Advanced; Glycosylation; Hydroxides; Mass Spectrometry; Molecular Structure; Ovalbumin

Effective adjuvants in anti-tumour vaccine formulations are very important in the development of new-generation vaccines. In this study, two layered double hydroxide (LDH) nanomaterial forms, i.e. nanoparticles (NPs) and nanosheets (NSs), were synthesised and examined as adjuvants to provoke the immune responses for anti-tumour purpose. Immunogen ovalbumin (OVA) delivered by both nanomaterials induced much stronger humoral and cell-medicated immune responses, together with an immune stimulant (TLR9 ligand CpG), as evidenced by higher levels of IgG1, IgG2a and interferon-γ. By comparison, LDH NSs showed higher activity to promote specific antibody responses than LDH NPs but with a similar cell-mediated immune response. The mice immunised with OVA-CpG vaccines formulated with both nanomaterials showed stronger inhibition of the inoculated tumour growth and had a longer survival. Altogether, these data indicate that LDH NPs and NSs can be used as potential nanoadjuvants for efficient protein-based anti-tumour vaccines.

Topics: Adjuvants, Immunologic; Animals; Cancer Vaccines; Cell Line, Tumor; Cell Proliferation; Drug Compounding; Female; Hydroxides; Mice; Nanoparticles; Ovalbumin

Modulation of the immune response is an important step in the induction of protective humoral and cellular immunity against pathogens. In this study, we investigated the possibility of using a nanomaterial conjugated with the toll-like receptor (TLR) ligand CpG to modulate the immune response towards the preferred polarity. MgAl-layered double hydroxide (LDH) nanomaterial has a very similar chemical composition to Alum, an FDA approved adjuvant for human vaccination. We used a model antigen, ovalbumin (OVA) to demonstrate that MgAl-LDH had comparable adjuvant activity to Alum, but much weaker inflammation. Conjugation of TLR9 ligand CpG to LDH nanoparticles significantly enhanced the antibody response and promoted a switch from Th2 toward Th1 response, demonstrated by a change in the IgG2a:IgG1 ratio. Moreover, immunization of mice with CpG-OVA-conjugated LDH before challenge with OVA-expressing B16/F10 tumor cells retarded tumor growth. Together, these data indicate that LDH nanomaterial can be used as an immune adjuvant to promote Th1 or Th2 dominant immune responses suitable for vaccination purposes.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Antibody Formation; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Chemical Phenomena; CpG Islands; Disease Models, Animal; Drug Delivery Systems; Female; Hydroxides; Immunity, Cellular; Immunization; Immunoglobulin G; Mice; Mice, Inbred C57BL; Nanoparticles; Ovalbumin

Dextran, a water-soluble, biocompatible polymer of glucose, was modified at its hydroxyls with arylboronic esters to make it soluble in common organic solvents, allowing for the facile preparation of oxidation-sensitive dextran (Oxi-DEX) carrier microparticles. These particles were found to release their payload with a half-life of 36 min at 1 mM H2O2, which can be compared with a half-life of greater than 1 week in the absence of H2O2. When used in a model vaccine application, Oxi-DEX particles loaded with ovalbumin (OVA) increased the presentation to CD8+ T-cells 27-fold relative to OVA encapsulated in a classical vehicle not sensitive to oxidation. No presentation was observed from cells incubated with unencapsulated OVA. Additionally, Oxi-DEX was found to be nontoxic in preliminary in vitro cytotoxicity assays. Because it is easy to prepare, sensitive to biological oxidation, and biocompatible, this material may represent an attractive new platform for selective delivery applications.

Topics: Animals; Antigen Presentation; Coated Materials, Biocompatible; Dendritic Cells; Dextrans; Drug Carriers; HeLa Cells; Humans; Hydroxides; Mice; Ovalbumin; Oxidation-Reduction; Solubility; Solvents

The effect of the degree of adsorption after exposure to interstitial fluid on the immune response in mice to model vaccines containing ovalbumin, alpha casein or dephosphorylated alpha casein adsorbed to aluminum hydroxide adjuvant was studied. Ovalbumin and dephosphorylated alpha casein were adsorbed in the vaccine but were completely eluted when exposed to interstitial fluid for 4 h. The presence of aluminum hydroxide adjuvant in the vaccine produced immunopotentiation compared to a solution of the protein even though the protein desorbed rapidly upon subcutaneous administration. In contrast, alpha casein was completely adsorbed to aluminum hydroxide adjuvant in both the vaccine and upon exposure to interstitial fluid. Immunopotentiation by aluminum hydroxide adjuvant was also observed in this model vaccine compared to a solution of alpha casein. The results indicated that antigen presenting cells can take up desorbed antigen from interstitial fluid as well as antigen adsorbed to aluminum-containing adjuvants.

Topics: Adjuvants, Immunologic; Adsorption; Aluminum Hydroxide; Animals; Antigens; Caseins; Cattle; Extracellular Space; Female; Hydroxides; Injections, Subcutaneous; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphates; Phosphorylation; Vaccines