PURPOSE: Human papillomavirus (HPV) is a significant cause of cervical cancer-related deaths worldwide. Because HPV is a sexually transmitted mucosal pathogen, enhancement of antigen-specific mucosal immune response likely serves good strategy for vaccination. However, mucosal vaccines generally do not induce strong enough immune responses. Previously we proved that a bacterial flagellin, Vibrio vulnificus FlaB, induce strong antigen-specific immune responses by stimulating the Toll-like receptor 5. In this study, we tested whether FlaB could serve as an effective mucosal adjuvant for a peptide-based HPV preventive cancer vaccine. MATERIALS AND METHODS: Mice were intranasally administered with a mixture of FlaB and E6/E7 protective peptides in 5-day interval for a total of two times. Five-days after the last vaccination, cellular immune responses of the vaccinated mice were analyzed. Tumor growth was also observed after a subcutaneous implantation of TC-1 cells bearing E6/E7 antigens. RESULTS: Intranasal administration of the E6/E7 peptide mixture with FlaB elicited a strong antigen-specific cytotoxic T lymphocyte activity and antigen-specific interferon-gamma production from splenocytes and cervical lymph node cells. Furthermore, FlaB, as a mucosal adjuvant, conferred an excellent protection against TC-1 tumor challenge with high survival rates in E6/E7 immunized animals. CONCLUSION: These results indicate that FlaB can be a promising mucosal adjuvant for nasal HPV vaccine development.
Administration, Intranasal ; Animals ; Flagellin ; Humans ; Immunity, Cellular ; Immunity, Mucosal ; Immunization ; Interferon-gamma ; Lymph Nodes ; Lymphocytes ; Mice ; Peptides ; Survival Rate ; Toll-Like Receptor 5 ; Ursidae ; Vaccination ; Vaccines ; Vibrio vulnificus

OBJECTIVETo study the mechanism of prophylactic effects of nasal tolerance with a dual analogue (Lys262-Ala207) on experimental autoimmune myasthenia gravis (EAMG).

METHODSClinical and immunological changes were observed in Lewis rats administered with dual analogue Lys262-Ala207 nasally, to compare the effects between the rats with predetermined dosage of Lys262-Ala207 and control peptides at two different time points, before the day (Group A or C) or on the day (Group B or D) of immunization with acetylcholine receptor (AChR) in complete Freud's adjuvant for 10 consecutive days. The clinical scores was evaluated for 50 days post immunization. Numbers of MNC expressing IFN-gamma, IL-4 or IL-10 and CD4+ and/or CD25+ from lymph nodes were enumerated by flow cytometry. Proliferative response, expressed as stimulation index (SI), was suppressed in response to antigen-specific stimulation in the rats receiving dual analogue, as compared with the rats receiving saline buffer only.

RESULTSGroup A and group B of Lewis rats developed EAMG with reduced severity, as compared to the control groups. Number of cells synthesizing IFN-gamma, IL-4 or IL-10 decreased, whereas numbers of CD4+CD25+ cells increased in group A and B than those in the control groups. Proliferative response was suppressed in response to antigen-specific stimulations in the rats receiving dual analogue Lys262-Ala207.

CONCLUSIONSNasal administration with a dual analogue Lys262-Ala207 at two different time points, before the day and on the day of immunization, could delay symptoms of muscular weakness in EAMG rats, which was associated with suppression of immune function in AChR antigen-specific T cells and lay a scientific foundation for treatment of human MG with nasal dual analogue.

Administration, Intranasal ; Animals ; Antibody Affinity ; Autoantibodies ; immunology ; Dose-Response Relationship, Immunologic ; Female ; Immune Tolerance ; drug effects ; Immunity, Cellular ; Immunity, Mucosal ; Lymphocyte Activation ; Myasthenia Gravis ; immunology ; prevention & control ; Nasal Mucosa ; immunology ; Rats ; Rats, Inbred Lew

A recombinant replication-defective adenovirus expressing the major epitopes of porcine circovirus-2 (PCV-2) capsid protein (rAd/Cap/518) was previously constructed and shown to induce mucosal immunity in mice following intranasal delivery. In the present study, immune responses induced by intranasal immunization with a combination of rAd/Cap/518 and cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) were evaluated in mice. The levels of PCV-2-specific IgG in serum and IgA in saliva, lung, and intestinal fluids were significantly higher in the group immunized with rAd/Cap/518 and CpG ODN than animals immunized with rAd/Cap/518 alone. The frequencies of IL-2-secreting CD4+ T cells and IFN-gamma-producing CD8+ T cells were significantly higher in the combined immunization group than mice immunized with rAd/Cap/518 alone. The frequencies of CD3+, CD3+CD4+CD8-, and CD3+CD4-CD8+ T cells in the combined immunization group were similar to that treated with CpG ODN alone, but significantly higher than mice that did not receive CpG ODN. PCV-2 load after challenge in the combined immunization group was significantly lower than that in the phosphate-buffered saline placebo group and approximately 7-fold lower in the group treated with CpG ODN alone. These results indicate that rAd/Cap/518 combined with CpG ODN can enhance systemic and local mucosal immunity in mice, and represent a promising synergetic mucosal vaccine against PCV-2.
Adenoviridae/genetics/immunology ; Administration, Intranasal ; Animals ; Capsid Proteins/*genetics/immunology ; Circoviridae Infections/*immunology ; Circovirus/*genetics/immunology ; Epitopes/genetics/immunology ; Female ; Immunity, Mucosal/immunology ; Immunoglobulin A/blood/immunology ; Immunoglobulin G/blood/immunology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides/genetics ; Vaccines, Synthetic/genetics/immunology ; Viral Vaccines/administration & dosage/*genetics/immunology

Vaccination is an effective strategy to prevent infectious or immune related diseases, which has made remarkable contribution in human history. Recently increasing attentions have been paid to mucosal vaccination due to its multiple advantages over conventional ways. Subunit or peptide antigens are more reasonable immunogens for mucosal vaccination than live or attenuated pathogens, however adjuvants are required to augment the immune responses. Many mucosal adjuvants have been developed to prime desirable immune responses to different etiologies. Compared with pathogen derived adjuvants, innate endogenous molecules incorporated into mucosal vaccines demonstrate prominent adjuvanticity and safety. Nowadays, cytokines are broadly used as mucosal adjuvants for participation of signal transduction of immune responses, activation of innate immunity and polarization of adaptive immunity. Desired immune responses are promptly and efficaciously primed on basis of specific interactions between cytokines and corresponding receptors. In addition, some other innate molecules are also identified as potent mucosal adjuvants. This review focuses on innate endogenous mucosal adjuvants, hoping to shed light on the development of mucosal vaccines.
Adjuvants, Immunologic ; Animals ; Humans ; Immunity, Innate ; immunology ; Immunity, Mucosal ; immunology ; Vaccines ; administration & dosage ; immunology

AIDS Vaccines ; administration & dosage ; genetics ; immunology ; Animals ; HIV Infections ; immunology ; prevention & control ; virology ; HIV-1 ; genetics ; immunology ; Humans ; Immunity, Mucosal

BACKGROUND AND OBJECTIVES: Cholera toxin B (CTB) is an effective immunomodulating agent. Mucosal tolerance is a well recognized method for inducing tolerance, but large amounts of antigen is needed. The dosage required can be dramatically reduced through coupling the antigen to the CTB. This study aimed to examine whether mucosal administration of house dust mite coupled to CTB (HDM-CTB conjugate) would modulate specific type 1 hypersensitivity in the murine model of allergic rhinitis and to evaluate the prophylactic and long-term therapeutic effects. MATERIALS AND METHOD: C57BL/6 mice were sensitized with Dermatophagoides farinae extract. After administration of HDM-CTB conjugate, several parameters of allergic response were evaluated. RESULTS: After mucosal adminstration of HDM-CTB conjugate, allergic symptoms, eosinophilic infiltration into nasal mucosa, specific IgE, and the splenic T cell proliferation after Dermatophagoides farinae allergen challenge were suppressed. CONCLUSION: These findings show that HDM-CTB conjugate has an anti-allergic effect in the murine model of allergic rhinitis and suggest that house dust mite allergy can be modulated with mucosal administration of allergen coupled to CTB.
Administration, Mucosal ; Animals ; Cell Proliferation ; Cholera Toxin ; Dermatophagoides farinae ; Dust* ; Eosinophils ; Hypersensitivity ; Immunoglobulin E ; Immunotherapy ; Mice* ; Nasal Mucosa ; Pyroglyphidae ; Rhinitis*

BACKGROUND: The perfect preanesthetic medication and its ideal route of administration are still debated. Transmucosal administration of midazolam has been of interest because of the rapid, reliable onset of action, predictable effects and avoidance of injections. Because many medications are well absorbed from the mucosa, we conducted a randomized, prospective, blinded study to compare acceptance and efficacy of intranasal and sublingual administration of midazolam as a preanesthetic medication in children. METHODS: One hundred twenty eight patients aged 0.5-12year were stratified by age: 38 infants and toddlers, 0.5-3yr; 48 preschoolers, 3.1-7yr; and 42 school age, 7.1-12yr. They were randomized to received 0.2 mg/kg of midazolam in the nose or under the tongue. Hemoglobin oxygen saturation by pulse oximetry and sedation score were recorded before drug administration, at 2.5min intervals for 15min, at separation from parents and during induction with enflurane in O2. Retention time of sublingual drug and duration of crying were recorded. RESULTS: The incidence of crying at the time of administration of midazolam was greater following intranasal compared with sublingual administration(60% vs 17%, p<0.05). Within age groups, only infants and toddlers showed a significant difference in the incidence of crying between treatment groups. Significant changes in sedation occured in both groups from 2.5min after administration. CONCLUSIONS: Sublingual midazolam is better accepted than intranasal midazolam as a preanesthetic sedative in children.
Administration, Mucosal ; Administration, Sublingual ; Child ; Crying ; Enflurane ; Humans ; Hypnotics and Sedatives ; Incidence ; Infant ; Midazolam* ; Mucous Membrane ; Nose ; Oximetry ; Oxygen ; Parents ; Preanesthetic Medication* ; Premedication ; Prospective Studies ; Tongue

In order to elucidate the physiological basis for mucosal immunity of oral vaccination and to present the essential carrier of microparticles or nanoparticles used to investigate the orally delivered vaccine, the features of antigen presentation and mucosal immunereaction in gut-associated lymphoid tissues were analyzed. Considered the morphological and physiological barriers of the gastrointestinal tract, absorption and transport of particulates were further discussed. And the studies about particulate dosage forms for oral vaccine delivery were also summarized in this review. Peyer s patches and M-cells, involved in immunoregulation, are significant areas performing the critical role in oral vaccine. The applied vesicle of microparticles could overcome the barriers of gastrointestinal tract. Oral vaccination was endued with new connotation, especially the enhanced transport and immunization efficiencies promoted by the lectin anchored particles. In conclusion, oral vaccination mediated by particulate carrier via mucosal immune system, would contribute to the site-specific triggering and signal magnification. For vaccines, the prospects for the application of these promising carrier systems might have potential attraction for scientific research and commercial development.
Administration, Oral ; Animals ; Drug Delivery Systems ; methods ; Humans ; Immunity, Mucosal ; Intestinal Absorption ; Microspheres ; Nanoparticles ; Vaccination ; methods ; Vaccines ; administration & dosage ; immunology ; pharmacokinetics

BACKGROUNDMucosal immunity is important to defense against dental caries. To enhance mucosal immunity, a DNA vaccine mucosal delivery system was prepared by encapsulating anticaries DNA vaccine (plasmid pGJA-P/VAX) in chitosan under optimal conditions and the characteristics of the microparticles was investigated. Furthermore, the release properties and protective action of microparticles for plasmid were studied in vitro.

METHODSPlasmid loaded chitosan microparticles were prepared by complex coacervation. Three factors, concentration of DNA, sodium sulfate, and the chitosan/DNA ratios in complexes [better expressed as N/P ratio: the number of poly nitrogen (N) per DNA phosphate (P)] influencing preparation were optimized by orthogonal test. The characteristics of microparticles were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). DNA release rate of microparticles in similar gastro fluid (SGF) or similar intestinal fluid (SIF) at 37 degrees C was determined by ultraviolet spectrophotometry.

RESULTSHigh encapsulation efficiency (96.8%) was obtained with chitosan microparticles made under optimal conditions of 50 mmol/L Na2SO4, 200 microg/ml DNA and N/P ratio of 4. The size of particles was about 4 to 6 microm. The encapsulation process did not destroy the integrity of DNA. When incubated with SIL, after a release of about 10% in the first 60 minutes, no further DNA was released during the following 180 minutes. When incubated with SGL, the microparticles released a small burst (about 11%) in the first 60 minutes, and then slowly released at a constant, but different rate.

CONCLUSIONSThese chitosan microparticles showed suitable characteristics in vitro for mucosal vaccination and are therefore a promising carrier system for DNA vaccine mucosal delivery.

Animals ; CHO Cells ; Cell Survival ; Chitosan ; administration & dosage ; Cricetinae ; Drug Delivery Systems ; Electrophoresis, Agar Gel ; Immunity, Mucosal ; Particle Size ; Vaccines, DNA ; administration & dosage ; metabolism

Influenza virus is one of the major sources of respiratory tract infection. Due to antigenic drift in surface glycoproteins the virus causes annual epidemics with severe morbidity and mortality. Although hemagglutinin (HA) is one of the highly variable surface glycoproteins of the influenza virus, it remains the most attractive target for vaccine development against seasonal influenza infection because antibodies generated against HA provide virus neutralization and subsequent protection against the virus infection. Combination of recombinant adenovirus (rAd) vector-based vaccine and mucosal administration is a promising regimen for safe and effective vaccination against influenza. In this study, we constructed rAd encoding the globular head region of HA from A/Puerto Rico/8/34 virus as vaccine candidate. The rAd vaccine was engineered to express high level of the protein in secreted form. Intranasal or sublingual immunization of mice with the rAd-based vaccine candidates induced significant levels of sustained HA-specific mucosal IgA and IgG. When challenged with lethal dose of homologous virus, the vaccinated mice were completely protected from the infection. The results demonstrate that intranasal or sublingual vaccination with HA-encoding rAd elicits protective immunity against infection with homologous influenza virus. This finding underlines the potential of our recombinant adenovirus-based influenza vaccine candidate for both efficacy and rapid production.
Adenoviridae* ; Administration, Mucosal ; Animals ; Antibodies ; Head* ; Hemagglutinins* ; Immunization* ; Immunoglobulin A ; Immunoglobulin G ; Influenza Vaccines ; Influenza, Human* ; Membrane Glycoproteins ; Mice* ; Mortality ; Orthomyxoviridae* ; Respiratory Tract Infections ; Seasons ; Vaccination ; Viruses