Enhancement of Immune Responses Elicited by Nanovaccines through a Cross-Presentation Pathway.

Numerous studies have aimed to develop novel advanced vaccines, in part because traditional vaccines have been unsuccessful in preventing rapidly emerging and reemerging viral and bacterial infections. There is a need for an advanced vaccine delivery system to ensure the successful induction of humoral and cellular immune responses. In particular, the ability of nanovaccines to modulate intracellular antigen delivery by inducing exogenous antigens (loaded onto major histocompatibility complex class 1 molecules) in CD8+ T cells, the so-called cross-presentation pathway, has attracted a great deal of attention. Protection against viral and intracellular bacterial infections relies on cross-presentation. This review discusses the advantages, requirements, and preparation of nanovaccines, the cross-presentation mechanism, the several parameters affecting cross-presentation by nanovaccines, and future perspectives.

Inactivated Vibrio cholerae Strains That Express TcpA via the toxT-139F Allele Induce Antibody Responses against TcpA.

Cholera remains a major global public health problem, for which oral cholera vaccines (OCVs) being a valuable strategy. Patients, who have recovered from cholera, develop antibody responses against LPS, cholera toxin (CT), toxin-coregulated pilus (TCP) major subunit A (TcpA) and other antigens; thus, these responses are potentially important contributors to immunity against Vibrio cholerae infection. However, assessments of the efficacy of current OCVs, especially inactivated OCVs, have focused primarily on O-antigen-specific antibody responses, suggesting that more sophisticated strategies are required for inactivated OCVs to induce immune responses against TCP, CT, and other antigens. Previously, we have shown that the toxT-139F allele enables V. cholerae strains to produce CT and TCP under simple laboratory culture conditions. Thus, we hypothesized that V. cholerae strains that express TCP via the toxT-139F allele induce TCP-specific antibody responses. As anticipated, V. cholerae strains that expressed TCP through the toxT-139F allele elicited antibody responses against TCP when the inactivated bacteria were delivered via a mouse model. We have further developed TCP-expressing V. cholerae strains that have been used in inactivated OCVs and shown that they effect an antibody response against TcpA in vivo, suggesting that V. cholerae strains with the toxT-139F allele are excellent candidates for cholera vaccines.

Progress of Polysaccharide-Contained Polyurethanes for Biomedical Applications.

Polyurethane (PU) has been widely examined and used for biomedical applications, such as catheters, blood oxygenators, stents, cardiac valves, drug delivery carriers, dialysis devices, wound dressings, adhesives, pacemaker, tissue engineering, and coatings for breast implants due to its mechanical flexibility, high tear strength, biocompatibility, and tailorable foams although bio-acceptability, biodegradability and controlled drug delivery to achieve the desired properties should be considered. Especially, during the last decade, the development of bio-based PUs has raised public awareness because of the concern with global plastic waste for creating more environmentally friended materials. Therefore, it is desirable to discuss polysaccharide (PS)-contained PU for the wound dressing and bone tissue engineering among bio-based PUs because PS has several advantages, such as biocompatibility, reproducibility from the natural resources, degradability, ease of incorporation of bioactive agents, ease of availability and cost-effectiveness, and structural feature of chemical modification to meet the desired needs to overcome the disadvantages of PU itself by containing the PS into the PU.

In vitro photodynamic therapy of methylene blue-loaded acetyl resistant starch nanoparticles.

BACKGROUND: Combination therapies comprising multiple methods, such as photodynamic therapy have been applied to be complements chemotherapy as they increase the therapeutic efficiency by enabling the intelligent drug delivery to target sites by exposing the photosensitizer to light and activating it in the tumor tissue. This study evaluated in vitro photodynamic therapy of methylene blue (MB)-loaded acetyl resistant starch (ARS) nanoparticles (NPs). METHODS: ARS was synthesized by the reaction between resistant starch (RS) and acetic anhydride. MB-loaded ARS NPs and ARS NPs were prepared by a single emulsion method. Synthesized ARS was measured by NMR. Prepared ARS NPs and MB-loaded ARS NPs were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), X-ray diffraction, UV/Vis, and circular dichroism (CD). MB-loaded ARS NPs were treated in mouse colon cancer cells (CT-26) and they were treated under near-infrared (NIR) laser irradiation. RESULTS: Synthesis of ARS was confirmed by NMR and the degree of substitutions in the ARS was 7.1. The morphologies of ARS NPs observed by TEM were spherical shapes and the particle sizes of ARS NPs were 173.4 nm with a surface charge of - 17.24 mV. The d-spacing of ARS NPs was smaller than those of RS and the conformational changes of RS occurred by the formation of self-assembled polymeric NPs with induction of CD of the MB by chiral ARS NPs. The phototoxicity of CT-26 cells treated by MB-loaded ARS NPs dramatically decreased in a dose-dependent manner under NIR laser irradiation compared to free MB. CONCLUSION: This study demonstrated the ordered nanosized structures in the ARS NPs and conformational change from random coil structure of RS to alpha-helices one of ARS occurred and CD of the achiral MB was induced. The MB-loaded ARS NPs showed a higher generation of reactive oxygen species (ROS) in the CT-26 cells than free MB with the NIR laser irradiation and resulting in phototoxicity under irradiation.

Bacteriophage EK99P-1 alleviates enterotoxigenic Escherichia coli K99-induced barrier dysfunction and inflammation.

Bacteriophages, simply phages, have long been used as a potential alternative to antibiotics for livestock due to their ability to specifically kill enterotoxigenic Escherichia coli (ETEC), which is a major cause of diarrhea in piglets. However, the control of ETEC infection by phages within intestinal epithelial cells, and their relationship with host immune responses, remain poorly understood. In this study, we evaluated the effect of phage EK99P-1 against ETEC K99-infected porcine intestinal epithelial cell line (IPEC-J2). Phage EK99P-1 prevented ETEC K99-induced barrier disruption by attenuating the increased permeability mediated by the loss of tight junction proteins such as zonula occludens-1 (ZO-1), occludin, and claudin-3. ETEC K99-induced inflammatory responses, such as interleukin (IL)-8 secretion, were decreased by treatment with phage EK99P-1. We used a IPEC-J2/peripheral blood mononuclear cell (PBMC) transwell co-culture system to investigate whether the modulation of barrier disruption and chemokine secretion by phage EK99P-1 in ETEC K99-infected IPEC-J2 would influence immune cells at the site of basolateral. The results showed that phage EK99P-1 reduced the mRNA expression of ETEC K99-induced pro-inflammatory cytokines, IL-1ß and IL-8, from PBMC collected on the basolateral side. Together, these results suggest that phage EK99P-1 prevented ETEC K99-induced barrier dysfunction in IPEC-J2 and alleviated inflammation caused by ETEC K99 infection. Reinforcement of the intestinal barrier, such as regulation of permeability and cytokines, by phage EK99P-1 also modulates the immune cell inflammatory response.

Mucosal Vaccine Delivery Using Mucoadhesive Polymer Particulate Systems.

Vaccination has been recently attracted as one of the most successful medical treatments of the prevalence of many infectious diseases. Mucosal vaccination has been interested in many researchers because mucosal immune responses play part in the first line of defense against pathogens. However, mucosal vaccination should find out an efficient antigen delivery system because the antigen should be protected from degradation and clearance, it should be targeted to mucosal sites, and it should stimulate mucosal and systemic immunity. Accordingly, mucoadhesive polymeric particles among the polymeric particles have gained much attention because they can protect the antigen from degradation, prolong the residence time of the antigen at the target site, and control the release of the loaded vaccine, and results in induction of mucosal and systemic immune responses. In this review, we discuss advances in the development of several kinds of mucoadhesive polymeric particles for mucosal vaccine delivery.

Galectin-9 Induced by Dietary Probiotic Mixture Regulates Immune Balance to Reduce Atopic Dermatitis Symptoms in Mice.

Probiotics can be an effective treatment for atopic dermatitis (AD), while their mechanism of action is still unclear. Here, we induced AD in mice with 2,4-dinitrochlorobenzene and administrated YK4, a probiotic mixture consisting of Lactobacillus acidophilus CBT LA1, L. plantarum CBT LP3, Bifidobacterium breve CBT BR3, and B. lactis CBT BL3. Then, we have validated the underlying mechanism for the alleviation of AD by YK4 from the intestinal and systematic immunological perspectives. Administration of YK4 in AD mice alleviated the symptoms of AD by suppressing the expression of skin thymic stromal lymphopoietin and serum immunoglobulin E eliciting excessive T-helper (Th) 2 cell-mediated responses. YK4 inhibited Th2 cell population through induce the proportion of Th1 cells in spleen and Treg cells in Peyer's patches and mesenteric lymph node (mLN). CD103+ dendritic cells (DCs) in mLN and the spleen were significantly increased in AD mice administered with YK4 when compared to AD mice. Furthermore, galectin-9 was significantly increased in the gut of AD mice administered with YK4. In vitro experiments were performed using bone marrow-derived DCs (BMDC) and CD4+ T cells to confirm the immune mechanisms of YK4 and galectin-9. The expression of CD44, a receptor of galectin-9, together with programmed death-ligand 1 was significantly upregulated in BMDCs following treatment with YK4. IL-10 and IL-12 were upregulated when BMDCs were treated with YK4. Cytokines together with co-receptors from DCs play a major role in the differentiation and activation of CD4+ T cells. Proliferation of Tregs and Th1 cell activation were enhanced when CD4+T cells were co-cultured with YK4-treated BMDCs. Galectin-9 appeared to contribute at least partially to the proliferation of Tregs. The results further suggested that DCs treated with YK4 induced the differentiation of naïve T cells toward Th1 and Tregs. At the same time, YK4 alleviated AD symptoms by inhibiting Th2 response. Thus, the present study suggested a potential role of YK4 as an effective immunomodulatory agent in AD patients.