Development and Pre-Clinical Evaluation of Recombinant Human Myelin Basic Protein Nano Therapeutic Vaccine in Experimental Autoimmune Encephalomyelitis Mice Animal Model.
Development and Pre-Clinical Evaluation of Recombinant Human Myelin Basic Protein Nano Therapeutic Vaccine in Experimental Autoimmune Encephalomyelitis Mice Animal Model.
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DOI：
10.1038/srep46468
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发表时间：
2017-04-20
影响因子：
4.6
通讯作者：
Laible G
中科院分区：
文献类型：
--
作者：
Al-Ghobashy MA;ElMeshad AN;Abdelsalam RM;Nooh MM;Al-Shorbagy M;Laible G
重组人髓磷脂碱性蛋白（rhMBP）以前在转基因奶牛的牛奶中生产。证明了表面固定化抗hMBP抗体对hMBP或rhMBP的分子识别的差异。这表明rhMBP和hMBP之间的免疫应答存在差异。在此，在实验性自身免疫性脑脊髓炎（EAE）动物模型中证明了游离和控释rhMBP聚（ε-己内酯）纳米颗粒（NPs）作为针对多发性硬化（MS）的治疗性疫苗的活性。通过优化纳米处方，得到了具有高包封率和控释模式的离散球形、表面粗糙的rhMBP纳米粒。结果表明，rhMBP被负载并静电吸附在纳米粒表面。在EAE诱导前皮下注射游离的或rhMBP NPs降低了EAE小鼠的平均行为评分，并且仅显示出轻微的组织学改变和髓鞘的保存，rhMBP NPs显示出增加的保护作用。此外，对小鼠脑中的炎性细胞因子（IFN-γ和IL-10）的分析显示，用游离或rhMBP NPs预处理显著保护免于诱导的炎症。最后：i）rhMBP改善了EAE动物模型中的EAE症状，ii）纳米制剂显著增强了rhMBP作为治疗性疫苗的功效，以及iii）需要临床研究来证明rhMBP NP作为MS的治疗性疫苗的活性。
Recombinant human myelin basic protein (rhMBP) was previously produced in the milk of transgenic cows. Differences in molecular recognition of either hMBP or rhMBP by surface-immobilized anti-hMBP antibodies were demonstrated. This indicated differences in immunological response between rhMBP and hMBP. Here, the activity of free and controlled release rhMBP poly(ε-caprolactone) nanoparticles (NPs), as a therapeutic vaccine against multiple sclerosis (MS) was demonstrated in experimental autoimmune encephalomyelitis (EAE) animal model. Following optimization of nanoformulation, discrete spherical, rough-surfaced rhMBP NPs with high entrapment efficiency and controlled release pattern were obtained. Results indicated that rhMBP was loaded into and electrostatically adsorbed onto the surface of NPs. Subcutaneous administration of free or rhMBP NPs before EAE-induction reduced the average behavioral score in EAE mice and showed only mild histological alterations and preservation of myelin sheath, with rhMBP NPs showing increased protection. Moreover, analysis of inflammatory cytokines (IFN-γ and IL-10) in mice brains revealed that pretreatment with free or rhMBP NPs significantly protected against induced inflammation. In conclusion: i) rhMBP ameliorated EAE symptoms in EAE animal model, ii) nanoformulation significantly enhanced efficacy of rhMBP as a therapeutic vaccine and iii) clinical investigations are required to demonstrate the activity of rhMBP NPs as a therapeutic vaccine for MS.