The results of the SNT for PI-3 and BRSV are presented Fludarabine in vivo in Fig. 2A and B. At the initial examination (week −2), the mean log titre of PI-3 and BRSV specific antibodies of all calves from both groups was 1.7 ± 0.08

and 0.45 ± 0.07, respectively. The PI-3 SNT profile indicated a comparable antibody profile between both groups, with no significant difference identified (p = 0.18). Antibody levels increased rapidly for both groups at week 3 p.i. (one week following 1st vaccination) and remained elevated with minor variations throughout. The BRSV SNT profile also indicated no differences between the groups (p = 0.62). Antibody titres fluctuated similarly for both groups, with rising titres mainly correlating with the vaccination protocol with the exception of low values at week 9. ELISA results showed all animals were positive for PI-3 antibodies (cut-off value at 10 PP) with a mean value of 109.8 ± 6.7 PP and 36 (75%) were positive for BRSV antibodies (cut-off value at 10 PP) with a mean value of 56.3 ± 7.1 PP prior to the vaccination. ELISA results for PI-3 are presented in Fig. 2C and these indicate that antibody titres were not significantly different between the groups, and the profile of antibody changes over time were identical. There

was an increase in antibody levels following primary vaccination (week 2 p.i.), with a steady decline following week 8. The ELISA results for BRSV (Fig. 2D) indicate increasing antibody click here levels following primary vaccination (week 2 p.i.), a steady increase to week 10 and then a decline. There was no difference between groups in either case, with a p-value of 0.81 for BRSV and 0.55 for PI-3. Antibody titres specific to M. haemolytica are presented in Fig. 2E, which shows there was no difference between the groups (p = 0.26). Animals from both groups showed high antibody titres on the day of vaccination, which increased over time with a peak at week 10. Levels of IgG2 were lower in comparison to IgG1 for both PI-3 (Fig. 3A) and BRSV

Thiamine-diphosphate kinase (Fig. 3B). There was a significant difference (p = 0.01) between the groups for BRSV IgG1 antibodies at week 4 but the difference was not evident thereafter. Levels of IgG1 for BRSV reached their peak at week 9, whereas levels of IgG1 for PI-3 peaked at week 4 and gradually declined thereafter. On the other hand, levels of IgG2 for BRSV were lowest at week 4 and at week 2 for PI-3 but no group effect was identified for any of the IgG2 levels. In vitro cytokine analysis indicated that the production of IFN-γ by PBMC was significantly higher (p = 0.048 for the overall model) in calves from the control group when the cell culture was not stimulated with antigens (Med) as shown in Fig. 4A. At week 8, IL-4 production was significantly higher (p = 0.03) in the control group from unstimulated cells ( Fig. 4B). At week 12 the concentration of IL-4 was significantly higher (p = 0.04) in the control group from cells stimulated with TLRL 7/8 ( Fig.