Su Fang-Yi, Zhao Qingyang Henry, Dahotre Shreyas N, Gamboa Lena, Bawage Swapnil Subhash, Silva Trenkle Aaron D, Zamat Ali, Phuengkham Hathaichanok, Ahmed Rafi, Santangelo Philip J, Kwong Gabriel A
The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30317, USA.
Sci Adv. 2022 Feb 25;8(8):eabm7950. doi: 10.1126/sciadv.abm7950. Epub 2022 Feb 23.
Simultaneous delivery of mRNA to multiple populations of antigen (Ag)-specific CD8 T cells is challenging given the diversity of peptide epitopes and polymorphism of class I major histocompatibility complexes (MHCI). We developed Ag-presenting nanoparticles (APNs) for mRNA delivery using pMHCI molecules that were refolded with photocleavable peptides to allow rapid ligand exchange by UV light and site-specifically conjugated with a lipid tail for postinsertion into preformed mRNA lipid nanoparticles. Across different TCR transgenic mouse models (P14, OT-1, and Pmel), UV-exchanged APNs bound and transfected their cognate Ag-specific CD8 T cells equivalent to APNs produced using conventionally refolded pMHCI molecules. In mice infected with PR8 influenza, multiplexed delivery of UV-exchanged APNs against three immunodominant epitopes led to ~50% transfection of a VHH mRNA reporter in cognate Ag-specific CD8 T cells. Our data show that UV-mediated peptide exchange can be used to rapidly produce APNs for mRNA delivery to multiple populations of Ag-specific T cells in vivo.
鉴于肽表位的多样性和I类主要组织相容性复合体（MHCI）的多态性，将信使核糖核酸（mRNA）同时递送至多个抗原（Ag）特异性CD8 T细胞群体具有挑战性。我们开发了用于mRNA递送的抗原呈递纳米颗粒（APN），其使用与光可裂解肽重折叠的pMHCI分子，以允许通过紫外线进行快速配体交换，并与脂质尾进行位点特异性缀合，以便在插入预先形成的mRNA脂质纳米颗粒后进行后插入。在不同的T细胞受体转基因小鼠模型（P14、OT-1和Pmel）中，紫外线交换的APN与使用传统重折叠的pMHCI分子产生的APN一样，能够结合并转染其同源Ag特异性CD8 T细胞。在感染PR8流感的小鼠中，针对三种免疫显性表位的紫外线交换APN的多重递送导致同源Ag特异性CD8 T细胞中约50%的VHH mRNA报告基因转染。我们的数据表明，紫外线介导的肽交换可用于快速生产APN，以便在体内将mRNA递送至多个Ag特异性T细胞群体。
