[Protective activity of Immunovac-VP-4 vaccine against avian influenza virus H5N2 administered by different methods].
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AIM To experimentally assess protective effect of Immunovac-VP-4 vaccine against avian influenza virus H5N2. MATERIALS AND METHODS. Immunization of mice with polycomponent vaccine Immunovac-VP-4 was performed using oral or mucosal route of administration (intranasally, orally, and with combined nasal-oral method). Immunized mice were inoculated intranasally by influenza virus H5N2 adapted for mice. Survival of mice in experimental and control (intact) groups was assessed daily during 14 days. Survival and death rates of mice were determined. Levels of cytokines in sera of mice from both groups were measured by enzyme immunoassay. RESULTS Half of experimental animals survived after triple subcutaneous administration of vaccine in dose 20 mcg and subsequent intranasal challenge with avian influenza virus H5N2. Single subcutaneous immunization with dose 400 mcg resulted in survival of 80 +/- 12.6% of mice after challenge. Triple intranasal and combined intranasal-oral immunization as well as after triple subcutaneous immunization resulted in survival of half of challenged mice. In control group challenge was lethal for 90 - 100% of mice. Same methods of immunization lead to increase of IL-6, IL-12, IL-15, and IFN-gamma levels. CONCLUSION Data about significant protective effect after immunization with Immunovac-VP-4 against avian influenza virus H5N2 were obtained. Immunovac-VP-4 administered by mentioned routes activated nasal-associated lymphoid tissue providing first line defense at entry site of influenza infection, which demonstrates need to further study of this vaccine during development of strategy for non-specific prophylaxis of influenza infection.Keywords:
Subcutaneous injection
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The synthetic peptide GK-1, derived from Taenia crassiceps, enhances the protection induced by human influenza vaccine in both young and aged mice. Herein, the adjuvant properties of GK-1 fused to the pVIII protein of a heat-inactivated phagemid vector (FGK1) when co-administered with the influenza vaccine were assessed, to evaluate its feasibility as a low-cost adjuvant. In mice, FGK1 significantly increased the expected IgG and IgA anti-influenza antibody levels both in sera and in bronchoalveolar fluids when intranasally or subcutaneously co-administered with influenza vaccine. Single-dose pig co-immunization with FGK1 and influenza vaccine induced serum levels of IgG anti-influenza antibodies similar to those elicited by a two-dose immunization with the influenza vaccine alone. Preclinical evaluation of FGK1 with the influenza vaccine is currently in progress, in order to recommend its use for veterinary purposes.
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NMRI mice were immunized intramuscular, intranasal or by Aerosol, using the ethylethylenimine inactivated and polyethylenglycolconcentrated influenza virus strain A/PR/8/34 (HO/N1). Differences in the immune response resulted from all three routes. Intranasal and intramuscular vaccination were superior to aerosol application. A possible explanation for this could be the fact that relatively small amounts of the inhaled virus antigen developed antigenic activity on the mucous membrane. A single vaccination by the aerosol technique gave significant protection only, if the challenge virus was applied by the same procedure. However no protection was found after intranasal challenge. Intranasal challenge on the third day post vaccination revealed that intramuscular immunization had a significant better protective effect than intranasal immunization. However from the 5th to the 10th day post vaccination this effect reversed and intranasal vaccination became superior. This immunity persisted for the whole period of observation and it was accompained by a higher titer of local antibodies. Similar results were obtained in experiments with aerosol challenge. Here only the intranasal vaccinated mice were completely protected after the 10th day post vaccinationem while intramuscular vaccinated animals were less protected. Sera of intramuscular immunized mice revealed a higher content in antibodies of the Ig M type and less of the Ig G type compared to mice vaccinated by the intranasal route.
Intramuscular injection
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Avian influenza subunit vaccines have been shown to be poorly immunogenic, leading to the re-evaluation of the immunogenic and dose-sparing potential of whole virus vaccines. In this study, we investigated the immune responses after one or two doses of intramuscular or intranasal whole inactivated influenza H5N1 virus vaccine in BALB/c mice. Serum samples and nasal washings were collected weekly post-vaccination and analysed using enzyme-linked immunosorbent assay (ELISA). Sera were also analysed by the haemagglutination inhibition (HI) assay. Antibody-secreting cells were measured in lymphocytes from spleen and bone marrow via enzyme-linked immunospot (ELISPOT). Splenocytes were stimulated in vitro, and T-helper profiles were measured through multiplex bead assay in the supernatants, or intracellularly by multiparametric flow cytometry. Both vaccine routes induced high HI titres following the second immunization (intramuscular = 370, intranasal = 230). Moreover, the intramuscular group showed significantly higher levels of serum IgG (P < 0.01), IgG1 (P < 0.01) and IgG2a (P < 0.01) following the second vaccine dose, while the intranasal group exhibited significantly higher levels of serum IgA (P < 0.05) and local IgA (P < 0.01) in the nasal washings. Also, IgA antibody-secreting cells were found in significantly higher numbers in the intranasal group in both the spleen (P < 0.01) and the bone marrow (P < 0.01). Moreover, Th1 (TNF-α, IL-2, IFN-γ) and Th2 (IL-4, IL-5, IL-10) cytokines were expressed by both groups, yet only the intranasal group expressed the Th17 marker IL-17. As the intranasal vaccines induce local IgA and are easily administered, we suggest the intranasally administered whole virus vaccine as a promising candidate for a pandemic H5N1 vaccine.
ELISPOT
BALB/c
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Hemagglutination assay
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Inactivated vaccine
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Hemagglutination assay
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Two split influenza virus vaccines administered intragastrically induced lower titres of haemagglutinin (HA)-specific antibodies in pulmonary secretions than whole virus vaccine or a third split virus vaccine. IgA antibody was the predominant HA-specific Ig class. HA-specific IgA titres decayed substantially within 2 weeks following booster immunization, but persisted for at least another 3.5 months. In contrast, HA-specific IgG was maintained at low titres throughout the 4 month study period. When the total vaccine antigenic mass was administered as one dose or as equally divided doses spread over several days, pulmonary antibody responses were comparable. Mice immunized intragastrically with whole virus vaccine were completely protected against intranasal challenge with a homologous virulent virus of the H3 subtype. Partial protection was obtained when the vaccine used for immunization was a distantly related, antigenically variant strain of the same subtype, but no protection was obtained with a monovalent vaccine of an influenza A subtype different to the challenge virus. These characteristics of the response to intragastric immunization against influenza are consistent with features of a useful vaccine.
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In 2009, a global epidemic of influenza A(H1N1) virus caused the death of tens of thousands of people. Vaccination is the most effective means of controlling an epidemic of influenza and reducing the mortality rate. In this study, the long-term immunogenicity of influenza A/California/7/2009 (H1N1) split vaccine was observed as long as 15 months (450 days) after immunization in a mouse model. Female BALB/c mice were immunized intraperitoneally with different doses of aluminum-adjuvanted vaccine. The mice were challenged with a lethal dose (10× 50% lethal dose [LD(50)]) of homologous virus 450 days after immunization. The results showed that the supplemented aluminum adjuvant not only effectively enhanced the protective effect of the vaccine but also reduced the immunizing dose of the vaccine. In addition, the aluminum adjuvant enhanced the IgG antibody level of mice immunized with the H1N1 split vaccine. The IgG level was correlated to the survival rate of the mice. Aluminum-adjuvanted inactivated split-virion 2009 pandemic influenza A H1N1 vaccine has good immunogenicity and provided long-term protection against lethal influenza virus challenge in mice.
Inactivated vaccine
Pandemic
Lethal dose
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