HRV-specific IFN-γ producing T cell responses in the Gn pigs vaccinated with AttHRV with or without LA. Mononuclear cells from the ileum, spleen and peripheral blood of pigs were extracted and assayed on PID 28. Frequencies of IFN-γ+CD4+ and IFN-γ+CD8+ T cells among CD3+ MNC were determined by an intracellular staining and flow cytometry assay after the MNC were stimulated with purified AttHRV antigen for 17 h. Frequencies from the mock-stimulated MNC were subtracted from the frequencies of the AttHRV-stimulated MNC. Data represent the adjusted mean frequency (n = 4 for LA+AttHRV+; n = 7 for LA−AttHRV+) of HRV-specific IFN-γ producing T cells. Bars with different letters (A and B) on top differ significantly for the same tissue (Kruskal–Wallis Test, p < 0.05).

(a and b) HRV-specific ASC and total Ig SC responses in Gn pigs vaccinated with AttHRV with or without LA. Mononuclear cells from the ileum, spleen and peripheral blood of pigs were extracted and assayed on PID 28. Enzyme-linked-immunospot assays for determining HRV-specific ASC and total IgSC numbers were performed on the day of MNC extraction. Data represent the mean numbers of HRV-specific ASC (a) and total IgSC (b) per 5 × 10⁵ MNC, respectively (n = 4–7). Bars with different letters (A–C) on top differ significantly among groups for the same tissue and the same antibody isotype (Kruskal–Wallis Test, p < 0.05).

(a and b) Geometric mean titers (GMT) of isotype-specific antibodies to HRV and total Ig GMT in Gn pigs vaccinated with AttHRV with or without LA. Data represent the GMT of HRV-specific antibody (a) and total Ig (b) determined by ELISA at each time point (n = 7–9). Data points on lines marked with different letters (A–D) differ significantly (one-way ANOVA followed by Duncan’s multiple range test on log₁₀ transferred titers, p = 0.05).