Choi Jin Huk, Schafer Stephen C, Freiberg Alexander N, Croyle Maria A
†Division of Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, Texas 78712, United States.
‡Department of Pathology, The University of Texas Medical Branch, Galveston, Texas 77555, United States.
Mol Pharm. 2015 Aug 3;12(8):2697-711. doi: 10.1021/mp5006454. Epub 2015 Jan 20.
The severity and longevity of the current Ebola outbreak highlight the need for a fast-acting yet long-lasting vaccine for at-risk populations (medical personnel and rural villagers) where repeated prime-boost regimens are not feasible. While recombinant adenovirus (rAd)-based vaccines have conferred full protection against multiple strains of Ebola after a single immunization, their efficacy is impaired by pre-existing immunity (PEI) to adenovirus. To address this important issue, a panel of formulations was evaluated by an in vitro assay for their ability to protect rAd from neutralization. An amphiphilic polymer (F16, FW ∼39,000) significantly improved transgene expression in the presence of anti-Ad neutralizing antibodies (NAB) at concentrations of 5 times the 50% neutralizing dose (ND50). In vivo performance of rAd in F16 was compared with unformulated virus, virus modified with poly(ethylene) glycol (PEG), and virus incorporated into poly(lactic-co-glycolic) acid (PLGA) polymeric beads. Histochemical analysis of lung tissue revealed that F16 promoted strong levels of transgene expression in naive mice and those that were exposed to adenovirus in the nasal cavity 28 days prior to immunization. Multiparameter flow cytometry revealed that F16 induced significantly more polyfunctional antigen-specific CD8+ T cells simultaneously producing IFN-γ, IL-2, and TNF-α than other test formulations. These effects were not compromised by PEI. Data from formulations that provided partial protection from challenge consistently identified specific immunological requirements necessary for protection. This approach may be useful for development of formulations for other vaccine platforms that also employ ubiquitous pathogens as carriers like the influenza virus.
当前埃博拉疫情的严重程度和持续时间凸显了为高危人群(医务人员和农村村民)研发一种起效快且持久的疫苗的必要性,因为在这些人群中重复的初免-加强免疫方案并不可行。虽然基于重组腺病毒(rAd)的疫苗单次免疫后能对多种埃博拉病毒株提供完全保护,但其效力会受到对腺病毒的预先存在免疫(PEI)的影响。为解决这一重要问题,通过体外试验评估了一组制剂保护rAd免受中和的能力。一种两亲性聚合物(F16,分子量约39,000)在抗腺病毒中和抗体(NAB)浓度为50%中和剂量(ND50)的5倍时,能显著提高转基因表达。将F16中的rAd的体内性能与未配制的病毒、用聚乙二醇(PEG)修饰的病毒以及包封于聚乳酸-羟基乙酸共聚物(PLGA)聚合物微球中的病毒进行了比较。肺组织的组织化学分析显示,F16在未接触过腺病毒的小鼠以及在免疫前28天经鼻腔接触过腺病毒的小鼠中均能促进高水平的转基因表达。多参数流式细胞术显示,与其他测试制剂相比,F16诱导同时产生IFN-γ、IL-2和TNF-α的多功能抗原特异性CD8+T细胞显著更多。这些效应不受PEI的影响。来自能提供部分攻毒保护的制剂的数据一致确定了保护所需的特定免疫要求。这种方法可能有助于开发其他疫苗平台的制剂,这些平台也采用普遍存在的病原体作为载体,如流感病毒。