Journal:

Article Title: Subunit Rotavirus Vaccine Administered Parenterally to Rabbits Induces Active Protective Immunity

doi:

Figure Lengend Snippet: Serum and intestinal antibody responses following two parenteral immunizations (0 and 21 or 28 dpv) of rabbits with formalin-inactivated SA11 rotavirus, MA104 cell lysate, or 50 μg of G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in Freund’s adjuvants (A to C) and with 50 μg of live/psoralen-inactivated SA11 rotavirus, TNC buffer, or 50 μg of G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in QS-21 (D to F). Total (IgM, IgG, and IgA) serum (A and D), intestinal IgG (B and E), and intestinal IgA (C and F) antirotavirus antibodies were measured by ELISA before (56 or 69 dpv) and after (84 or 97 dpv) oral challenge with lapine ALA rotavirus. Antibody titers were measured for individual rabbits, and results are plotted as GMTs of the groups. The GMT of intestinal IgG antibody titers for both 0 and 28 dpc of the rabbits vaccinated with 2/6-VLPs in Freund’s adjuvants is 368. Therefore, for this group, only one box appears on the graph (B). Error bars represent 1 standard error of the mean. For titers of <50 and <5, 25 and 2.5, respectively, were used to calculate the GMT. A GMT of 25 or 2.5, for serum or intestinal antibodies, respectively, was considered negative. A significant (P < 0.05, Wilcoxon signed ranks test) increase in rotavirus antibody GMTs following challenge is indicated by an asterisk.

Article Snippet: Vaccine preparations were either (i) adsorbed to AlP (kindly supplied by Wyeth Lederle Vaccines and Pedriatics, West Henrietta, N.Y.) as described previously ( 19 ); (ii) mixed 1:1 with Freund’s adjuvants (complete for the first dose and incomplete for the second dose); or (iii) mixed with 20 μg (unless otherwise specified) of QS-21 adjuvant (kindly supplied by Wyeth Lederle Vaccines and Pediatrics), a purified triterpene glycoside (saponin) of aquillaic acid derivative from the bark of the South American Molina tree Quillaja saponaria ( 45 , 46 ).

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Subunit Rotavirus Vaccine Administered Parenterally to Rabbits Induces Active Protective Immunity

doi:

Figure Lengend Snippet: Protection from lapine ALA rotavirus challenge in rabbits immunized twice parenterally with formalin-inactivated SA11 rotavirus, MA104 cell lysate, or 50 μg of G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in Freund’s adjuvant (A) or with 50 μg of live/psoralen-inactivated SA11 rotavirus, TNC buffer, or 50 μg of G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in QS-21 (B). Virus antigen shedding curves were plotted for individual rabbits, and the mean area under the curve calculated for each group. Results are plotted as percent reduction in shedding for each individual animal (•), as well as the mean for each vaccine group (—), calculated by comparing the mean area under the curve for each vaccine group to the mean area under the curve for the control group. The number of rabbits with overlapping levels of protective efficacy is indicated next to the dots (n = 2 or 3); formulation of vaccine and number of rabbits per group are indicated along the x axis.

Article Snippet: Vaccine preparations were either (i) adsorbed to AlP (kindly supplied by Wyeth Lederle Vaccines and Pedriatics, West Henrietta, N.Y.) as described previously ( 19 ); (ii) mixed 1:1 with Freund’s adjuvants (complete for the first dose and incomplete for the second dose); or (iii) mixed with 20 μg (unless otherwise specified) of QS-21 adjuvant (kindly supplied by Wyeth Lederle Vaccines and Pediatrics), a purified triterpene glycoside (saponin) of aquillaic acid derivative from the bark of the South American Molina tree Quillaja saponaria ( 45 , 46 ).

Techniques:

Journal:

Article Title: Subunit Rotavirus Vaccine Administered Parenterally to Rabbits Induces Active Protective Immunity

doi:

Figure Lengend Snippet: Serum immune responses to individual rotavirus proteins before (56 or 69 dpv) or after (84 or 97 dpv) oral challenge with lapine ALA rotavirus. Representative results are shown for rabbits vaccinated parenterally either with 2/4/6/7-VLPs in Freund’s adjuvants (F) or with 2/4/6/7-, 2/6/7-, or 2/6-VLPs, psoralen-inactivated SA11, or TNC buffer in QS-21 (Q). Proteins in a SA11-infected cell lysate were separated by SDS-PAGE, transferred to nitrocellulose, and incubated with MAbs directed to VP4, VP6, or VP7, with a hyperimmune serum to SA11, or with paired sera from rabbits immunized with the indicated vaccine formulations. Percent protection from infection for individual rabbits is indicated below each lane. Locations of the viral proteins are indicated on the left.

Article Snippet: Vaccine preparations were either (i) adsorbed to AlP (kindly supplied by Wyeth Lederle Vaccines and Pedriatics, West Henrietta, N.Y.) as described previously ( 19 ); (ii) mixed 1:1 with Freund’s adjuvants (complete for the first dose and incomplete for the second dose); or (iii) mixed with 20 μg (unless otherwise specified) of QS-21 adjuvant (kindly supplied by Wyeth Lederle Vaccines and Pediatrics), a purified triterpene glycoside (saponin) of aquillaic acid derivative from the bark of the South American Molina tree Quillaja saponaria ( 45 , 46 ).

Techniques: Infection, SDS Page, Incubation

Journal:

Article Title: Subunit Rotavirus Vaccine Administered Parenterally to Rabbits Induces Active Protective Immunity

doi:

Figure Lengend Snippet: Correlation of higher protection rates with presence of serum N-Abs to VP4. (A) Serum N-Ab responses following two parenteral immunizations (0 and 28 dpv) of rabbits with 50 μg of P[2], G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in Freund’s adjuvants or with 50 μg of live or psoralen-inactivated SA11, TNC buffer, or 50 μg of P[2], G3 2/4/6/7-, G3 2/6/7-, or 2/6-VLPs in QS-21 were measured by FFNA against rotavirus strains SA11 (P[2], G3) and B223 (P8[11], G10) and against reassortant virus strains R-N33 (P8[11], G3) and R-A32 (P[2], G10) prior to (56 or 69 dpv) oral challenge with lapine ALA rotavirus. Serum N-Ab titers were measured for individual rabbits, and results are plotted as the GMTs of the groups. Error bars represent 1 standard error of the mean. For titers of <100, 50 was used to calculate the GMT. A GMT of 50 and its log were considered negative. A significant difference (P ≤ 0.021, Mann-Whitney U test) in VP4-specific N-Abs induced by 2/4/6/7-VLPs vaccines measured by comparing serum N-Abs measured against B223 and R-A32 is indicated by an asterisk. (B) Scatterplot of the log of the SA11 VP4-specific-NAb titers and the percent reduction of virus antigen shedding for individual rabbits (n = 58). Results were graphed to indicate whether rabbits were immunized with both VP4 and VP7 (■; SA11 or 2/4/6/7-VLPs in either adjuvant), absence of VP4 but presence of VP7 (•; 2/6/7-VLPs in either adjuvant), or absence of both VP4 and VP7 (▴; 2/6-VLPs in either adjuvant or TNC buffer in QS-21). Correlation coefficient was calculated by Pearson’s correlation coefficient.

Article Snippet: Vaccine preparations were either (i) adsorbed to AlP (kindly supplied by Wyeth Lederle Vaccines and Pedriatics, West Henrietta, N.Y.) as described previously ( 19 ); (ii) mixed 1:1 with Freund’s adjuvants (complete for the first dose and incomplete for the second dose); or (iii) mixed with 20 μg (unless otherwise specified) of QS-21 adjuvant (kindly supplied by Wyeth Lederle Vaccines and Pediatrics), a purified triterpene glycoside (saponin) of aquillaic acid derivative from the bark of the South American Molina tree Quillaja saponaria ( 45 , 46 ).

Techniques: MANN-WHITNEY