ABSTRACT
Objective:To investigate the optimal dose of Th1/Th2 immuno-enhancement effects of cultivated Artemisia rupestris L. crude polysaccharides (CARCP) on foot-and-mouth disease vaccine (FMDV) via the intramuscular route. Methods:ICR mice were intramuscularly immunized twice with different concentrations of CARCP mixed with FMDV at 2-week intervals. FMDV-specific antibodies, isotypes, and IgE in serum were detected by ELISA. Splenocyte proliferation was detected by MTT. T lymphocyte subsets and cytokines in the spleen were detected by flow cytometry. Clinical signs and local reactions at the injection site were monitored daily, and the body weight of mice was weighed after immunization.Results:The medium dose of CARCP could significantly improve FMDV-specific IgG, IgG 1, and IgG 2a antibody levels and the IgG 2a/IgG 1 ratio ( P<0.05) and lead to significant splenocyte proliferative responses ( P<0.01). The medium dose of CARCP could also significantly increase the level of CD3 +CD4 + and CD3 +CD8 + T cells as well as CD4 +/CD8 + ratio ( P<0.05), elicited the higher levels of IFN-γ in CD4 + T cells and CD8 + T cells ( P<0.05). No local adverse reactions at the injection site were observed after immunization. There was no significant difference in body weight or growth between each group( P>0.05). CARCP did not significantly induce an IgE response ( P>0.05). Conclusions:CARCP as an FMDV adjuvant promotes Th1/Th2 immune responses, especially in favor of the Th1 response, and has a certain safety profile. The best immune enhancement is achieved with CARCP at medium doses.
ABSTRACT
We developed a pre-treatment method to remove interfering substances during quantification of 146S antigens in foot-and-mouth disease (FMD) vaccines by high performance size exclusion chromatography (HPSEC). Three methods, including ultracentrifugation, PEG precipitation and nuclease digestion, were optimized and compared for removal efficiency of the interfering impurities in FMD vaccines. Under optimized conditions, the 146S contents in two batches of FMD vaccines were determined to be 7.1 and 7.6 μg/mL by ultracentrifugation, 9.7 and 10.4 μg/mL by PEG precipitation, and 10.5 and 10.4 μg/mL by nuclease digestion. The optimal condition for nuclease digestion using Benzonase determined by response surface method was as follows: appending Benzonase into 200 μL of antigen phase to a final concentration of 421 U/mL and incubating at 25.1 °C for 1.29 h. This method has advantages including efficient removal of the interfering impurities, fast processing speed, and mild operating conditions. Then 12 bathes of FMD vaccines with different serotypes produced by 4 manufacturers were tested to verify the established treatment method. Results showed the method was applicable to various FMD vaccines with good reproducibility (RSD<5.3%, n=3). The developed method removed interference from impurities during quantification of 146S, and therefore would broaden the application of HPSEC in vaccine quality control and ensure the accuracy and reliability.
Subject(s)
Animals , Chromatography, Gel , Foot-and-Mouth Disease , Foot-and-Mouth Disease Virus , Reproducibility of Results , Viral VaccinesABSTRACT
This study investigated changes in certain blood parameters in calves and pigs after foot-and-mouth disease (FMD) vaccination. In this study, five calves and five pigs were selected from groups of 10 calves and pigs, respectively, and were vaccinated with an FMD vaccine. The remaining animals formed two non-treatment control groups. Blood samples were collected from all animals on the 1st, 3rd, 5th, and 7th days post-vaccination. In the FMD-vaccinated calves and pigs on day 7 post-vaccination, white blood cell counts, blood urea nitrogen levels, and alanine aminotransferase and aspartate aminotransferase activities were higher than those in the respective controls. The present data suggested that the certain hemato-biochemical parameters on cattle and pigs were meaningfully changed between before and after FMD vaccination.