Karevan Ghazal, Ahmadi Kazem, Taheri Ramezan Ali, Fasihi-Ramandi Mahdi
Department of Biology, Nourdanesh Institute of Higher Education, Meymeh, Iran.
Molecular Biology Research Center, System Biology and Poisoning Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Clin Exp Vaccine Res. 2021 Jan;10(1):35-43. doi: 10.7774/cevr.2021.10.1.35. Epub 2021 Jan 31.
Brucellosis as a worldwide zoonotic illness affect domestic animals and humans doesn't have any vaccine for the prevention of infection in humans yet. The aim of this study was to evaluate the specific immune response following the administration of glycine nanoparticles as adjuvant and delivery system of a chimeric antigen contained trigger factor, Omp31, and Bp26 in murine model.
The chimeric antigen of was cloned and expressed in () BL21 (DE3). Purification and characterization of recombinant protein was conducted through Ni-NTA (nickel-nitrilotriacetic acid) agarose, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), and Western blot. Nanoparticle characteristics including morphology, particle size distribution, zeta potential, protein retention rate, and release rate were measured . Subsequently, nanoparticle contained antigen was administered to mice and blood sample was taken to measured the antibody level.
The protein retention in the nanoparticles was successfully done and the nanoparticle characteristics were appropriate. The average size of glycine particles containing antigen was about 174 nm, and the absorption of protein was approximately 61.27% of the initial value, with a release rate of approximately 70% after 8 hours. Enzyme-linked immunosorbent assay result proved that the immunized sera of mice which were administered with nano-formula contains high levels of antibodies (immunoglobulin G) against recombinant chimeric antigen and also a high level of mucosal antibody (immunoglobulin A) in the oral group, which showed a desirable immunity against .
The results showed that chimeric antigen-loaded glycine nanoparticles can act as a vaccine candidate for inducing the cellular and humoral immune response against brucellosis.
布鲁氏菌病作为一种全球性人畜共患病,影响家畜和人类,目前尚无预防人类感染的疫苗。本研究的目的是在小鼠模型中评估甘氨酸纳米颗粒作为包含触发因子、外膜蛋白31(Omp31)和Bp26的嵌合抗原的佐剂和递送系统给药后的特异性免疫反应。
将嵌合抗原克隆并在大肠杆菌(Escherichia coli)BL21(DE3)中表达。通过镍-次氮基三乙酸(Ni-NTA)琼脂糖、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹法对重组蛋白进行纯化和表征。测量纳米颗粒的特性,包括形态、粒径分布、zeta电位、蛋白质保留率和释放率。随后,将含抗原的纳米颗粒给予小鼠,并采集血样以测量抗体水平。
纳米颗粒中蛋白质保留成功,纳米颗粒特性合适。含抗原的甘氨酸颗粒平均大小约为174 nm,蛋白质吸收约为初始值的61.27%,8小时后释放率约为70%。酶联免疫吸附测定结果证明,用纳米配方给药的小鼠免疫血清含有高水平的针对重组嵌合抗原的抗体(免疫球蛋白G),并且口服组中黏膜抗体(免疫球蛋白A)水平也很高,这显示出对布鲁氏菌病的理想免疫力。
结果表明,负载嵌合抗原的甘氨酸纳米颗粒可作为诱导针对布鲁氏菌病的细胞免疫和体液免疫反应的候选疫苗。