Carnauba wax nanoparticles enhance strong systemic and mucosal cellular and humoral immune responses to HIV-gp140 antigen

Induction of humoral responses to HIV at mucosal compartments without inflammation is important for vaccine design. We developed charged wax nanoparticles that efficiently adsorb protein antigens and are internalized by DC in the absence of inflammation. HIV-gp140-adsorbed nanoparticles induced stronger in vitro T-cell proliferation responses than antigen alone. Such responses were greatly enhanced when antigen was co-adsorbed with TLR ligands. Immunogenicity studies in mice showed that intradermal vaccination with HIV-gp140 antigen-adsorbed nanoparticles induced high levels of specific IgG. Importantly, intranasal immunization with HIV-gp140-adsorbed nanoparticles greatly enhanced serum and vaginal IgG and IgA responses. Our results show that HIV-gp140-carrying wax nanoparticles can induce strong cellular/humoral immune responses without inflammation and may be of potential use as effective mucosal adjuvants for HIV vaccine candidates.

Multilayered hydrogel coatings covalently-linked to glass surfaces showing a potential to mimic mucosal tissues

Multilayered hydrogel coatings can be developed on the surface of glass slides via layer-by-layer deposition of hydrogen-bonded interpolymer complexes formed by poly(acrylic acid) and methylcellulose. Chemical modification of the glass surface with (3-aminopropyl)triethoxysilane with subsequent layer-by-layer deposition and cross-linking of interpolymer complexes by thermal treatment allows fabrication of ultrathin hydrogel coatings, not detachable from the substrate. The thickness of these coatings is directly related to the number of deposition cycles and cross-linking conditions. An unusual dependence of the hydrogel swelling properties on the sample thickness is observed and can be interpreted by gradual transitions between two- and three-dimensional networks. The hydrogels exhibit pH-responsive swelling behaviour, achieving higher swelling degrees at pH > 6.0. These coatings can be used as model substrates to study the adhesive properties of pharmaceutical tablets and can potentially mimic the total work of adhesion observed for the detachment of mucoadhesives from porcine buccal mucosa but fail to exhibit identical detachment profiles.

The long-term effect of active immunization against inhibin in goats

This experiment addresses the long-term effect of active immunization of goats against a recombinant ovine inhibin alpha subunit (roIHN-alpha). In late anestrus 100 mu g of roINH-alpha was administered to 40 pluriparous Boer goat does, followed, 4 weeks later, by a booster injection. Weekly blood samples were drawn to monitor the inhibin binding capacity with the aid of a radio-tracer binding assay. From the onset until 48 h after the end of each estrus, follicular development and ovulation rate were monitored at 24 h intervals by transrectal ultrasonography. Beginning in August and continuing into January, does were mated at every other estrus, and submitted to transcervical embryo collection. Seven months after the first immunization, the does were mated again and permitted to carry to term. All immunized does produced inhibin antibodies, an elevated titre being first detected 2 weeks after primary immunization. Maximum titres were reached after 6 weeks, i.e. 2 weeks after the booster injection. Thereafter, in the course of the following 32 weeks, the titre subsided gradually. The does started cycling by mid-August. At that stage the average number of follicles more than 4 mm in diameter, ovulations and total embryos and ova recovered were 14.7 (+/- 2.3), 5.3 (+/- 0.7) and 4.4 (+/- 1.0), respectively. A steady decline followed and in January the corresponding means were: 5.2 (+/- 0.6) follicles, 3.1 (+/- 0.6) ovulations and 1.2 (+/- 0.4) embryos and ova recovered. When mated toward the end of the breeding season, 85% of the does became pregnant to the first mating and 73% went to term. Healthy kids were born, the average litter size being 2.2 (+/- 0.1). In conclusion, immunization of goats against a recombinant inhibin alpha-subunit proved to be a practicable means of producing embryos for transfer purposes. After about half a year, when the inhibin antibody titre has subsided, it is possible to return the does to the breeding flock without risking complications with normal breeding activity. (c) 2009 Elsevier Inc. All rights reserved.

Effect of fecal water on an in vitro model of colonic mucosal barrier function

Fecal water (FW) has been shown to exert, in cultured cells, cytotoxic and genotoxic effects that have implications for colorectal cancer (CRC) risk. We have investigated a further biological activity of FW, namely, the ability to affect gap junctions in CACO2 cell monolayers as an index of mucosal barrier function, which is known to be disrupted in cancer. FW samples fi-om healthy, free-living, European subjects that were divided into two broad age groups, adult (40 +/- 9.7 yr; n = 53) and elderly (76 +/- 7.5 yr; n = 55) were tested for effects on gap junction using the transepithelial resistance (TER) assay. Overall, treatment of CACO2 cells with FW samples fi-om adults increased TER (+ 4 %), whereas FW from elderly subjects decreased TER (-5%); the difference between the two groups was significant (P < 0.05). We also measured several components of FW potentially associated with modulation of TER, namely, short-chain fatty acid (SCFA) and ammonia. SCFAs (propionic, acetic, and n-butyric) were significantly lower in the elderly population (-30%, -35%, and -21%, respectively, all P pound 0.01). We consider that FW modulation of in vitro epithelial barrier function is a potentially useful noninvasive biomarker, but it requires further validation to establish its relationship to CRC risk.

Adjuvant-free immunization with hemagglutinin-Fc fusion proteins as an approach to influenza vaccines

The hemagglutinins (HAs) of human H1 and H3 influenza viruses and avian H5 influenza virus were produced as recombinant fusion proteins with the human immunoglobulin Fc domain. Recombinant HA-human immunoglobulin Fc domain (HA-HuFc) proteins were secreted from baculovirus-infected insect cells as glycosylated oligomer HAs of the anticipated molecular mass, agglutinated red blood cells, were purified on protein A, and were used to immunize mice in the absence of adjuvant. Immunogenicity was demonstrated for all subtypes, with the serum samples demonstrating subtype-specific hemagglutination inhibition, epitope specificity similar to that seen with virus infection, and neutralization. HuFc-tagged HAs are potential candidates for gene-to-vaccine approaches to influenza vaccination.

Postnatal development of the mucosal immune system and consequences on health in adulthood

The intestinal microbiota is a dynamic multifaceted ecosystem which has evolved a complex and mutually beneficial relationship with the mammalian host. The contribution to host fitness is evident, but in recent years it has become apparent that these commensal microorganisms may exert far more influence over health and disease than previously thought. The gut microbiota are implicated in many aspects of biological function, such as metabolism, angiogenesis and immune development: disruption, especially during the neonatal period, which may impose life-long penalty. Elimination of the microbiota appears difficult, but manipulation of the ratios and dominance of composite populations can be achieved by alterations in diet, rearing environment, antibiotics and/or probiotics. Components of the intestinal microbiota are frequently documented to affect normal function of the mucosal immune system in experimental animals and in domesticated, agricultural species. However, it is not always clear that the effects described are sufficiently well understood to provide a sound basis for commercial intervention. Some microbial interventions may be beneficial to the host under particular circumstances, while detrimental during others. It is essential that we further our understanding of the complex and intricate host-commensal relationship to avoid causing more long-term damage than advantage

POZylation: a new approach to enhance nanoparticle diffusion through mucosal barriers

The increasing use of nanoparticles in the pharmaceutical industry is generating concomitant interest in developing nanomaterials that can rapidly penetrate into, and permeate through, biological membranes to facilitate drug delivery and improve the bioavailability of active pharmaceutical ingredients. Here, we demonstrate that the permeation of thiolated silica nanoparticles through porcine gastric mucosa can be significantly enhanced by their functionalization with either 5 kDa poly(2-ethyl-2-oxazoline) or poly(ethylene glycol). Nanoparticle diffusion was assessed using two independent techniques; Nanoparticle Tracking Analysis, and fluorescence microscopy. Our results show that poly(2-ethyl-2-oxazoline) and poly(ethylene glycol) have comparable abilities to enhance diffusion of silica nanoparticles in mucin dispersions and through the gastric mucosa. These findings provide a new strategy in the design of nanomedicines, by surface modification or nanoparticle core construction, for enhanced transmucosal drug delivery.