Synthesis of bursin and hyaluronic acid graft copolymer and its immune-enhancing activity
To enhance the immune effect of Bursin(BS),we attempted to graft BS to Hyaluronic acid.The amino groups of BS was utilized for direct coupling with the carboxylic groups on hyaluronic acid to obtain BS@HA by EDC and NHS.The structure of the polymer was identified by Fourier transform infrared spectroscopy(FTIR),quantitative analysis was performed using UV measurement.Then the mice were immunized with the vaccine formulated with simulated antigen OVA,BS@HA and aluminum hydroxide adjuvant and detected the immune enhancing effect by ELISA and flow cytometry.The results showed that:1)The yield of the synthesized polymer could reach 95％when the molar ratio of hyaluronic acid,EDC,NHS,and BS was 1∶4∶6∶0.4,the reaction time was 2-4 hours and 16 hours after adding EDC and NHS respectively.The FTIR characteristic peaks of HA shifting from 1 605 cm-1to 1 632 cm-1 showed BS@HA was successfully prepared.2)BS@HA can induce higher expression of OVA-specific antibodies when the degrees of substitution(DS)was about 35％.3)BS@HA also can induce higher percentages of Follicular helper T cells and germinal center B cells.Therefore,it is spected BS@HA could enhance targeted delivery of immunogens to draining lymph nodes,induce more TFH and GC B cells and evoke high antigen-specific IgG titers.This study will enrich the theoretical research of hyaluronic acid in the field of animal vaccines,expand its practical application in the animal vaccines and lay the foundation for the development of novel immune enhancers.
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