Chitosan-HPMC-blended microspheres as a vaccine carrier for the delivery of tetanus toxoid
Chitosan-HPMC-blended microspheres as a vaccine carrier for the delivery of tetanus toxoid
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DOI：
10.3109/21691401.2014.966193
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发表时间：
2016-02-17
影响因子：
5.8
通讯作者：
Jang, Hyun Tae
中科院分区：
文献类型：
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作者：
Arthanari, Saravanakumar;Mani, Ganesh;Jang, Hyun Tae
本研究的目的是开发一种合适的破伤风类毒素（TT）疫苗的替代佐剂，在单次免疫后诱导长期免疫。在我们的研究中，通过使用不同比例（7/3、8/2和9/1）的壳聚糖-羟丙基甲基纤维素（HPMC）混合空微球进行处方前研究。此外，TT用肝素（肝素浓度为1%、2%、3%和4%w/v）稳定，并通过水包油包水（W/O/W）多重乳化法包封在理想的壳聚糖- HPMC（CHBMS）微球中。在90天的时间内评价CHBMS的疫苗包封和体外释放效率。通过ELISA测定抗原从微球的释放。通过SDS-PAGE研究抗原完整性。从优化研究中发现，壳聚糖/HPMC的比例为8/2产生了良好的产率，微球为球形，规则且大小均匀。在CHBMS中，3%w/v的肝素浓度导致良好持续的抗原递送达90天。结果表明，该缓释剂在2 d内具有初释、恒释和90 d内几乎100%完全释药的特征。根据体外释放特性，通过抗体诱导方法评价了用于体内研究的CHBMS（3% w/v肝素）的理想批次。在9个月的时间内测量不同组合的抗体水平，最后在1年后进行第二次加强剂量。总之，观察到CHBMS（组合-1）导致豚鼠血清抗体水平为4.5 IU/mL，中央研究所的明矾吸附破伤风类毒素（CRITT）（组合2）在第二次加强剂量1年后的水平为3.5 IU/mL。这种使用CHBMS的新方法可能具有疫苗单步免疫的潜在优势。
The purpose of this research was to develop a suitable and alternate adjuvant for the tetanus toxoid (TT) vaccine that induces long term immunity after a single-dose immunization. In our study, the preformulation studies were carried out by using different ratios (7/3, 8/2, and 9/1) of chitosan-hydroxypropyl methylcellulose (HPMC)-blended empty microspheres. Moreover, TT was stabilized with heparin (at heparin concentrations of 1%, 2%, 3%, and 4% w/v) and encapsulated in ideal chitosan - HPMC (CHBMS) microspheres, by the water-in-oil-in-water (W/O/W) multiple emulsion method. The vaccine entrapment and the in vitro release efficiency of the CHBMS was evaluated for a period of 90 days. The release of antigens from the microspheres was determined by ELISA. Antigen integrity was investigated by SDS-PAGE. From the optimization studies, it was found that a chitosan/HPMC ratio of 8/2 produced a good yield, with microspheres that were spherical, regular and uniformly-sized. In the CHBMS, a heparin concentration of 3% w/v resulted in well-sustained antigen delivery for a period of 90 days. It was found that the characteristics of initial release could be observed in 2 days, followed by a constant release, and an almost 100% complete release in 90 days. From the in vitro release characteristics, the ideal batch of CHBMS (3% w/v heparin) was evaluated for in vivo studies by the antibody induction method. The antibody levels were measured for different combinations for the period of 9 months, and finally, with a second booster dose after 1 year. In conclusion, it was observed that CHBMS (combination-1) resulted in the antibody level of 4.5 IU/mL of guinea pig serum, and the level was 3.5 IU/mL for the Central Research Institute's alum-adsorbed tetanus toxoid (CRITT) (combination 2), after 1 year, with a second booster dose. This novel approach of using CHBMS may have potential advantages for single-step immunization with vaccines.