Influence of particle size, antigen load, dose and additional adjuvant on the immune response from antigen loaded PLA microparticles

Polylactide (PLA) polymer particles entrapping tetanus toxoid (TT) were evaluated in terms of particle size, antigen load, dose and additional adjuvant for achieving high and sustained anti-TT antibody titer from single point intramuscular immunization. Admixture of polymer entrapped TT and alum improved the immune response in comparison to particle-based immunization. High and long lasting antibody titer was achieved upon immunization with 2-8 microm size particles. Microparticles within the size range 50-150 microm elicited very low serum antibody response. Immunization with very small particles (<2 microm) and with intermediate size range particles (10-70 microm) elicited comparable antibody response from single point immunization but lower in comparison to that achieved while immunizing with 2-8 microm size particles. Potentiation of antibody response on immunization of admixture of microparticles and alum was also dependent on particle size. These results indicate the need of optimal particle sizes in micron ranges for improved humoral response from single point immunization. Increasing antigen load on polymer particles was found to have positive influence on generation of antibody titers from particle based immunization. Maximum peak antibody titer of approximately 300 microg/mL was achieved on day 50 upon immunization with particles having highest load of antigen (94 microg/mg of polymer). Increase in dose of polymer entrapped antigen resulted in concomitant increase in peak antibody titers indicating the importance of antigen stability, particle size and load on generating reproducible immune response. Optimization of particle size, antigen load, dose and use of additional adjuvant resulted in high and sustained anti-TT antibody titers over a period of more than 250 days from single point immunization. Serum anti-TT antibody titers from single point immunization of admixrure of PLA particles and alum was comparable with immunization from two divided doses of alum adsorbed TT.