Live attenuated Salmonella typhimurium vaccines delivering SaEsxA and SaEsxB via type III secretion system confer protection against Staphylococcus aureus infection.

BACKGROUND: Staphylococcus aureus (S. aureus) causes a wide range of infectious diseases in human and animals. The emergence of antibiotic-resistant strains demands novel strategies for prophylactic vaccine development. In this study, live attenuated S. enterica subsp. enterica serotype Typhimurium (S. Typhimurium) vaccine against S. aureus infection was developed, in which Salmonella Pathogenesis Island-1 Type 3 Secretion System (SPI-1 T3SS) was employed to deliver SaEsxA and SaEsxB, two of ESAT-6-like (Early Secreted Antigenic Target-6) virulence factors of S. aureus. METHODS: Antigens SaEsxA and SaEsxB were fused with the N-terminal secretion and translocation domain of SPI-1 effector SipA. And cytosolic delivery of Staphylococcal antigens into macrophages was examined by western blot. BALB/c mice were orally immunized with S. Typhimurium-SaEsxA and S. Typhimurium-SaEsxB vaccines. Antigen-specific humoral and Th1/Th17 immune responses were examined by ELISA and ELISPOT assays 7-9 days after the 2nd booster. For ELISPOT assays, the statistical significance was determined by Student's t test. The vaccine efficacy was evaluated by lethal challenge with two S. aureus clinical isolates Newman strain and USA 300 strain. Statistical significance was determined by Log rank (Mantel-Cox) analysis. And a P value of < 0.05 was considered statistically significant. RESULTS: Oral administration of S. Typhimurium-SaEsxA and S. Typhimurium-SaEsxB vaccines induced antigen-specific humoral and Th1/Th17 immune responses, which increased the survival rate for vaccinated mice when challenged with S. aureus strains. CONCLUSIONS: The newly developed S. Typhimurium-based vaccines delivering SaEsxA and SaEsxB by SPI-1 T3SS could confer protection against S. aureus infection. This study provides evidence that translocation of foreign antigens via Salmonella SPI-1 T3SS into the cytosol of antigen presenting cells (APCs) could induce potent immune responses against pathogens.

Strong Immune Responses Induced by Direct Local Injections of Modified mRNA-Lipid Nanocomplexes.

In vitro transcribed mRNAs hold the promises of many medical applications in disease prevention and treatment, such as replacement or supplement of missing or inadequately expressed endogenous proteins and as preventive vaccines against infectious diseases, therapeutic vaccines, or other protein-based biopharmaceutics for cancer therapy. A safe and efficient delivery system for mRNA is crucial to the success of mRNA therapeutic applications. In this study, we report that InstantFECT, a liposome-based transfection reagent, can pack pseudouridine-incorporated mRNA into nanocomplexes that are highly efficient in mediating in vivo transfection in multiple organs after local delivery. High levels of expression of EGFP and luciferase reporters after intratumoral and intramuscular injections were observed, which lasted for up to 96 hrs. Immunogenicity of antigens encoded by mRNA delivered with nanocomplex was investigated by subcutaneous delivery of modified mRNAs encoding Staphylococcus aureus adenosine synthase A (AdsA) and a model tumor-associated antigen ovalbumin (OVA). Strong T cell responses were provoked by both mRNAs delivered. Therapeutic and protective treatment with the OVA mRNA-liposome nanocomplex significantly inhibited B16-OVA tumor progression and increased mouse survival. There was no sign of obvious toxicity related to the treatment both in tissue culture and in mice. An intravenous injection of the same dosage of the modified mRNA-lipid nanocomplex showed minimal transfection in major organs, indicating an excellent safety feature as the gene transfer occurred only at the injection sites, whereas intravenous (i.v.) injection with the same amount of mRNA complexed with a commercial transfection reagent Trans-IT showed luciferase expression in the spleen. In summary, InstantFECT cationic liposomes provide a safe and efficient in vivo locoregional delivery of mRNA and could be a useful tool for basic research and for the development of mRNA-based therapies.

Hybrid nanovaccine for the co-delivery of the mRNA antigen and adjuvant.

For efficient cancer vaccines, the antitumor function largely relies on cytotoxic T cells, whose activation can be effectively induced via antigen-encoding mRNA, making mRNA-based cancer vaccines an attractive approach for personalized cancer therapy. While the liposome-based delivery system enables the systemic delivery and transfection of mRNA, incorporating an adjuvant that is non-lipid like remains challenging, although the co-delivery of mRNA (antigen) and effective adjuvant is key to the activation of the cytotoxic T cells. This is because the presence of an adjuvant is important for dendritic cell maturation-another necessity for cytotoxic T cell activation. In the present work, we designed a poly (lactic-co-glycolic acid) (PLGA)-core/lipid-shell hybrid nanoparticle carrier for the co-delivery of mRNA and gardiquimod (adjuvant that cannot be incorporated into the lipid shell). We demonstrated in the present work that the co-delivery of mRNA and gardiquimod led to the effective antigen expression and DC maturation in vitro. The intravenous administration of the hybrid nanovaccine resulted in the enrichment of mRNA expression in the spleen and a strong immune response in vivo. The simultaneous delivery of the antigen and adjuvant both spatially and temporally via the core/shell nanoparticle carrier is found to be beneficial for tumor growth inhibition.

Tongue abscess: a rare clinical entity.

Swelling of the tongue is a rare clinical entity. It is a potentially life-threatening condition, as it could result in airway compromise. The differential diagnosis of acute tongue swelling includes hemorrhage, infarction, abscess, tumor, and edema. A tongue abscess should be considered in all cases of acute tongue swelling, especially when host defenses are severely impaired. Although the diagnosis of lingual abscess can be reached clinically because of the rarity of the condition, in neglected cases, the diagnosis can be difficult. Despite of the rarity and complexity of this condition, its management strategy is relatively simple. In the present study, we describe a case of a tongue abscess on the anterior two-thirds of the tongue in a 60-year-old woman, and discuss the pathophysiology, diagnosis, and treatment of this complex entity.

Evaluation of surface radiation dose to the thyroid gland and the gonads during routine full-mouth intraoral periapical and maxillary occlusal radiography.

AIM: The quantitative aspects of radiation doses to critical organs can help the dental professionals to take the necessary radiation protective measures as deemed necessary and can help the general public to allay radiation exposure fear in dental radiography, if any. Our study determines the surface radiation dose to thyroid and gonads in full-mouth intraoral periapical (IOPA) and maxillary occlusal radiography. MATERIALS AND METHODS: A total number of 120 subjects participated in the study. The surface radiation dose was estimated to the thyroid gland and the gonads in full-mouth IOPA radiography using 10 IOPA (E speed films) and in maxillary occlusal radiography. The measurements were calculated using a digital pocket dosimeter (PD-4507). RESULTS: The average dose at the thyroid gland level during full-mouth intraoral and maxillary occlusal radiography was estimated to be 10.93 mRads (1.093 × 10(-2) mGy) and 0.4 mRads (4.0 × 10(-2) mGy), respectively. The average surface radiation dose at the gonadal region during a full mouth intraoral and maxillary occlusal radiography was estimated to be 1.5 mRads (1.5 × 10(-2) mGy) and 0.15 mRads (1.5 × 10(-3) mGy), respectively. CONCLUSION: Our results suggest that although the radiation exposure doses to critical organs namely thyroid and gonads is within the safe limits still precautionary measures for these organs are advocated.