Quintilio Wagner, Takata Célia S, Sant'Anna Osvaldo A, da Costa Maria Helena B, Raw Isaías
Quality Control Service, Instituto Butantan, Av. Dr Vital Brazil, 1500 SP 05503-900 Brazil.
Curr Drug Deliv. 2009 Jul;6(3):297-04. doi: 10.2174/156720109788680886.
Polymeric microspheres containing diphtheria and tetanus toxoids were prepared without protein stabilizers. A vaccine containing 2 Lf(tetanus) and 0.4 Lf(diphtheria) was injected either in BALB/c mice or in guinea-pigs. As control, a group received the alum-adsorbed unencapsulated toxoids. In mice, on day 44 one group and control received a booster and at day 111 the other group received the same booster dose. Before de booster, all groups had very low neutralizing antibodies, as determined by Toxin binding inhibition assay. One week after booster all groups had high antibody titers, especially those immunized with microencapsulated vaccine, which were at least 5 times higher than those immunized with alum vaccine for both antigens. Besides, guinea pigs receiving lower dose had antibodies titers as high as 60 UI/mL, and 30 times higher than those immunized with alum vaccine. Therefore by using an encapsulated vaccine without any kind of protein stabilizer we were able to induce in vivo protective responses irrespective of observed in vitro protein degradation by HPLC. Manipulating the vaccination schedule at the same time to the toxoids encapsulation does not only increase the antibody titers but also their specificity.
制备了不含蛋白质稳定剂的含有白喉和破伤风类毒素的聚合物微球。将含有2Lf（破伤风）和0.4Lf（白喉）的疫苗注射到BALB/c小鼠或豚鼠体内。作为对照，一组接受明矾吸附的未包封类毒素。在小鼠中，一组在第44天和对照组接受加强免疫，另一组在第111天接受相同的加强剂量。在加强免疫前，通过毒素结合抑制试验测定，所有组的中和抗体都非常低。加强免疫一周后，所有组的抗体滴度都很高，尤其是用微囊化疫苗免疫的组，两种抗原的抗体滴度至少比用明矾疫苗免疫的组高5倍。此外，接受较低剂量的豚鼠抗体滴度高达60 UI/mL，比用明矾疫苗免疫的组高30倍。因此，通过使用不含任何蛋白质稳定剂的包封疫苗，我们能够在体内诱导保护性反应，而不管通过高效液相色谱法观察到的体外蛋白质降解情况如何。同时调整疫苗接种时间表和类毒素包封不仅能提高抗体滴度，还能提高其特异性。
