Ontiveros-Padilla Luis, Hendy Dylan A, Pena Erik S, Williamson Grace L, Murphy Connor T, Lukesh Nicole R, Ashcraft Kathleen A, Abraham Mathew A, Landon Chelsea D, Staats Herman F, Abraham Soman N, Carlock Michael, Ross Ted M, Petrovsky Nikolai, Heise Mark T, Bachelder Eric M, Ainslie Kristy M
Division of Pharmacoengineering & Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA.
Division of Pharmacoengineering & Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA; Department of Biomedical Engineering, North Carolina State University and University of North Carolina, Chapel Hill, NC, USA.
J Control Release. 2025 Aug 10;384:113855. doi: 10.1016/j.jconrel.2025.113855. Epub 2025 May 18.
Flumist is the only FDA-approved intranasal influenza vaccine. Although it has recently been approved for at-home use, it has significant limitations. These include reduced effectiveness in generating a protective immune response in patients with extensive influenza exposure, safety concerns due to its live attenuated virus formulation, and reduced efficacy due to viral drift/shift. To address this limitation, we have developed a nanocomplex comprised of a mast cell (MC) agonist and toll-like receptor 9 (TLR9) ligand to adjuvant a broadly acting influenza antigen. The newly reported MC agonist was identified by screening mastoparan-7 analogs for MC degranulation activity, which led to a more active peptide analog, MP12W. Positively charged MP12W spontaneously forms nanoparticulate complexes (NPs) with CpG 1826 that were then used to intranasally vaccinate mice with a computationally optimized broadly reactive antigen (COBRA) hemagglutinin (HA) protein. The NPs were further optimized by substituting CpG 1826 with CpG 55.2, a TLR-9 agonist identified by machine learning to be more active in humans. MP12W-CpG 1826 NPs showed an increased pro-inflammatory response and decreased cytotoxicity in vitro compared to M7 complexes, translating into a safer profile in a model of increased hypersensitivity, collaborative cross mice 027 (CC027). Intranasal vaccination with this complex and broadly reactive HA resulted in higher mucosal antibody concentration and increased cytokine production with antigen recall. These responses were enhanced with MP12W-CpG 55.2 NP vaccination. MP12W-CpG NPs provided similar protection in an influenza challenge model. This study demonstrates the potential of this novel intranasal nanocomplex for vaccination.
弗卢米司特是美国食品药品监督管理局（FDA）批准的唯一一种鼻内流感疫苗。尽管它最近已被批准可在家中使用，但它有显著的局限性。这些局限性包括：在流感暴露广泛的患者中产生保护性免疫反应的有效性降低；由于其减毒活病毒制剂而存在安全问题；以及由于病毒漂移/变异导致效力降低。为了解决这一局限性，我们开发了一种纳米复合物，它由肥大细胞（MC）激动剂和Toll样受体9（TLR9）配体组成，用于辅助一种具有广泛作用的流感抗原。新报道的MC激动剂是通过筛选马蜂毒素-7类似物的肥大细胞脱颗粒活性而鉴定出来的，这导致了一种活性更高的肽类似物MP12W。带正电荷的MP12W与CpG 1826自发形成纳米颗粒复合物（NPs），然后用其对小鼠进行鼻内接种一种通过计算优化的具有广泛反应性的抗原（COBRA）血凝素（HA）蛋白。通过用CpG 55.2替代CpG 1826对NPs进行了进一步优化，CpG 55.2是一种通过机器学习鉴定出在人类中更具活性的TLR-9激动剂。与M�复合物相比，MP12W-CpG 18㔪 NPs在体外显示出增强的促炎反应和降低的细胞毒性，在超敏反应增强模型协作杂交小鼠027（CC027）中转化为更安全的特征。用这种复合物和具有广泛反应性的HA进行鼻内接种导致更高的黏膜抗体浓度，并在抗原再次刺激时增加细胞因子产生。用MP12W-CpG 55.2 NPs接种可增强这些反应。MP12W-CpG NPs在流感攻击模型中提供了类似的保护。这项研究证明了这种新型鼻内纳米复合物用于疫苗接种的潜力。
