Rafferty D E, Elfaki M G, Montgomery P C
Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Vaccine. 1996 Apr;14(6):532-8. doi: 10.1016/0264-410x(95)00196-8.
Biodegradable microparticles (MPs) were prepared to contain dinitrophenylated bovine serum albumin (DNP-BSA) and the cytokines interleukin (IL)-5 and IL-6. The conformational integrity of these MPs was examined by scanning electron microscopy. DNP-BSA was evaluated, postentrapment, by a bicinchoninic acid assay, SDS-PAGE, isoelectric focusing, Western blotting, ELISA and spectrophotometric analysis. The bioactivity of the ILs postentrapment was measured using bioassays. Ocular-topical (OT) and intraperitoneal (IP) administration of loaded MPs induced serum IgG, tear IgA and vaginal wash (VW) IgA responses which persisted up to 45 days post secondary immunization (P2o). OT and IP delivery elicited serum IgG and tear IgA responses up to 140 days post tertiary immunization (P3o). VW IgA responses persisted up to 45 days and 140 days P3o OT and IP delivery, respectively. Overall, the inclusion of cytokines in antigen containing MPs enhanced tear IgA antibody levels following OT delivery P2o, while elevated VW IgA responses occurred following IP delivery P2o and P3o. These data demonstrate that antigen/cytokine loaded MPs can potentiate long-term mucosal antibody responses at both target and distal effector sites as well as elicit circulating antibodies.
制备了可生物降解的微粒（MPs），其包含二硝基苯基化牛血清白蛋白（DNP-BSA）以及细胞因子白细胞介素（IL）-5和IL-6。通过扫描电子显微镜检查这些MPs的构象完整性。包封后，通过二辛可宁酸测定、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）、等电聚焦、蛋白质印迹法、酶联免疫吸附测定（ELISA）和分光光度分析对DNP-BSA进行评估。使用生物测定法测量包封后ILs的生物活性。负载MPs的眼局部（OT）和腹腔内（IP）给药诱导血清IgG、泪液IgA和阴道冲洗液（VW）IgA反应，这些反应在二次免疫后（P2o）持续长达45天。OT和IP给药在三次免疫后（P3o）长达140天引发血清IgG和泪液IgA反应。VW IgA反应在P3o时OT和IP给药分别持续长达45天和140天。总体而言，在含抗原的MPs中加入细胞因子可提高OT给药P2o后泪液IgA抗体水平，而在IP给药P2o和P3o后VW IgA反应升高。这些数据表明，负载抗原/细胞因子的MPs可增强靶部位和远端效应部位的长期粘膜抗体反应以及引发循环抗体。
