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1.
Figure 1

Figure 1. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Immunization scheme of candidate S-OIV H1N1 and avian H5N1 vaccine. BALB/c mice were immunized with different doses of the produced vaccine S-OIV H1N1 or NIBRG-14 H5N1. (A) PBS control, 0.001 μg, 0.01 μg, and 0.05 μg of S-OIV H1N1 or NIBRG-14 H5N1 vaccine; (B) PBS control, 0.01 μg, and 0.05 μg of S-OIV H1N1 or NIBRG-14 H5N1 with or without alum or MPLA adjuvant. About 3 weeks after immunization, the mice were challenged with 106 TCID50 of A/California/7/2009 H1N1 or 1000 TCID50 NIBRG-14 H5N1.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
2.
Figure 3

Figure 3. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Survival Rates and Body Weight Loss of immunized mice post challenge with A/California/7/2009 Virus. Mice (n = 4 to 7 per group) immunized with PBS, and 0.001 μg, 0.01 μg, and 0.05 μg of vaccine with or without adjuvant (alum or MPLA), followed by lethal challenge with 106 TCID50 of A/California/7/2009 virus intranasally. Following 14 days of observation, survival rates (A) and weight changes (B) in each group are shown.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
3.
Figure 2

Figure 2. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Adjuvant promoted strong protective immune responses to S-OIV H1N1 virus. To realize the least immunization dose of S-OIV and H5N1 vaccine needed for mice to raise sufficient protective immune responses to A/California/7/2009 virus, mice (n = 7 to 8 per group) immunized with PBS, 0.001 μg, 0.01 μg, or 0.05 μg of vaccine in the presence or absence of adjuvant. Three weeks after the single immunization, mice sera were evaluated for (A) H1N1 IgG, (B) H5N1 IgG, and (C) H1N1 IgG1 and IgG2a, and (D) H5N1 IgG1 and IgG2a immune responses to A/California/7/2009 virus. The data represent the mean titers ± SD (error bars) of antibodies in each group of animals.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
4.
Figure 6

Figure 6. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

MPLA as the adjuvant significantly reduced lung viral titer. Mice were vaccinated with S-OIV H1N1 or NIBRG-14 H5N1 vaccines with or without different adjuvants. For the challenge, 106 TCID50 of virus was inoculated into mice nasal cavities. Three days post challenge, mice lungs were homogenized and inoculated into MDCK cells for 48 h. Virus titers were evaluated using a micro-plaque assay. (A) H1N1 vaccination, H1N1 virus challenge, (B) H5N1 vaccination, H5N1 virus challenge, (C) H5N1 vaccination, H1N1 virus challenge, (D) H1N1 vaccination, H5N1 virus challenge. The TCID50 titers of the virus were calculated by the method of Reed and Muench [reference [24]. P < 0.05 indicated the significance the differences in viral titers between two groups of vaccinated mice.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
5.
Figure 5

Figure 5. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Adjuvant promoted IgG responses of mice to homologous and heterogeneous influenza H1N1 and H5N1 virus. To realize the least immunization dose of S-OIV and H5N1 vaccine needed for mice to raise sufficient protective immune responses to A/California/7/2009 H1N1 and NIBRG-14 H5N1 virus, mice (n = 3 to 4 per group) immunized with PBS, 0.01 μg, or 0.05 μg of vaccine with or without adjuvant. After 3 weeks of single-dose immunization, mice sera were used to evaluate the immune response type (A) H1N1 IgG to H1N1 virus, (B) H5N1 IgG to H5N1 virus, (C) H5N1 IgG to H1N1 virus, and (D) H1N1 IgG to H5N1 virus immune responses. The data represent the mean titers ± SD (error bars) of antibodies in each group of animals.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
6.
Figure 7

Figure 7. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Neutralization antibody response of immunized-mice post challenge with S-OIV H1N1 or NIBRG-14 H5N1 influenza virus. Neutralization antibodies of mice post vaccinated with S-OIV H1N1 or NIBRG-14 H5N1 vaccines were evaluated using a microneutralization assay. Mice sera (pre-immune as negative; A/California/7/2009 NIBSC 09/152 and antiserum to NIBRG-14 H5N1 HA as positive control) were mixed with viruses (100 TCID50 of NIBRG-121 H1N1 or NIBRG-14 H5N1 virus) at room temperature for 1 hour, and were inoculated into 96-well MDCK cells (1.5 × 104/ml). Experiments were performed following the WHO protocol for the microneutralization assay. (A) H1N1 vaccination, H1N1 neutralization antibody, (B) H5N1 vaccination, H5N1 neutralization antibody, (C) H5N1 vaccination, H1N1 neutralization antibody, (D) H1N1 vaccination, H5N1 neutralization antibody. Error bar is the standard deviation of six serum samples.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.
7.
Figure 4

Figure 4. From: Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination.

Adjuvant enhanced vaccine to elicit a hemagglutination inhibition (HAI) antibody response. Mice (n = 7 to 8 per group) were immunized with single different dose of S-OIV H1N1 or NIBRG-14 H5N1 influenza vaccine with or without alum or MPLA as the adjuvant. The titers of serum specific antibodies were evaluated using the hemagglutination inhibition (HAI) test. Data are representative of two separate experiments of H1N1 vaccine immunized mice sera (No sera from H5N1-immunized mice contained HAI to NIBRG-14 H5N1 virus using chicken red blood cells). Comparisons of HAI antibody titer in mice immunized with PBS, 0.001 μg, 0.1 μg, or 0.5 μg of HA in the produced vaccine with or without adjuvant. For comparison of HAI antibody titers, Student’s t test was used to examine the significance of differences between HAI positive rates (with HAI titer 3 40) of each vaccinated group and control group (mice immunized with PBS only). A P value of < 0.05 was considered significant. The star “★” indicates significant differences.

Hui-Tsu Lin, et al. J Biomed Sci. 2013;20(1):19-19.

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