Nanoparticle Delivery of DNA Vaccine Against Zika Virus

纳米颗粒递送抗寨卡病毒 DNA 疫苗

基本信息

批准号：
9379639
负责人：
Hai-Quan Mao
金额：
$ 20.43万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-19 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9379639
关键词：
Address Alexa594 Amino Acid Sequence Amino Acids Analysis of Variance Antibodies Antibody Formation Antibody titer measurement Antigen Presentation Antigen Presentation Pathway Antigen-Presenting Cells Antigens Asia B-Cell Activation Benchmarking Bone Marrow Brazil CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cells Centers for Disease Control and Prevention (U.S.)Chimera organism Cytotoxic T-Lymphocytes DNA DNA Vaccines DNA delivery Dendritic Cells Development Dose Drainage procedure Electroporation Engineering Enzyme-Linked Immunosorbent Assay Enzymes Epidemic Esters Europe Exhibits Flow Cytometry Frequencies Gene Expression Generations Glycolates Helper-Inducer T-Lymphocyte Histocompatibility Antigens Class II Immune response Immunoglobulin G Immunologic Memory Infection Injection of therapeutic agent Intramuscular Label Laboratories Life Link Lymph Node Drainage Lymphocyte MHC Class II Genes Mediating Medicine Membrane Membrane Proteins Methods Mus Neutralization Tests North America Ovalbumin Phosphate Buffer Physical condensation Polyethylene Glycols Polyethyleneimine Polymerase Chain Reaction Polypropylenes Process Production Protein Subunits Race Reporting Reproducibility Reverse Transcription Saline Serum Site South America Subunit Vaccines Sulfides Surface Properties T-Cell Activation Testing Th2 Cells Time Tissues Transfection Transmission Electron Microscopy Vaccination Vaccines World Health Organization Zika Virus Zika virus vaccine base copolymer cytokine disorder prevention env Gene Products fetal bovine serum fight against innovation light scattering long term memory lymph nodes lymphatic drainage nanoparticle neutralizing antibody next generation nonhuman primate novel strategies pathogen plasmid DNA response subcutaneous transgene expression uptake vaccine candidate vaccine delivery whole body imaging

项目摘要

PROJECT SUMMARY The objective of this project is to develop DNA nanoparticles with efficient delivery to the local draining lymph nodes (dLN) via subcutaneous (s.c.) administration to promote dendritic cell (DC) transfection, antigen presentation, and T cell activation in the lymph nodes, and to elicit robust antibody titers and immunological memory against Zika virus. As the Zika virus (ZIKV) epidemic in Brazil spread around south and north Americas, Europe, and Asia, teams around the world have been racing to develop ZIKV vaccines. DNA vaccines offer many advantages in terms of ease of production, excellent stability with long shelf life, multivalent capability, and fast development cycle. The first generation of vaccine candidates is built with the pre-membrane and envelope sequences from a Brazilian ZIKV strain as the dominant immunogen. They have been tested as naked plasmid DNA vaccines administered by intramuscular (i.m.) injection, and generated effective neutralizing antibodies in mice and nonhuman primates. However, naked DNA vaccine inherently has low efficiency to generate Th2 response and immune memory, and requires higher DNA dose due to the low abundance of antigen presenting cells (APCs) at the injection site. This study will directly address these challenges by combining two novel approaches to engineer a more potent ZIKV DNA vaccine: (1) a chimeric DNA construct encoding ZIKA envelope protein sequences and lysosomal-associated membrane protein-1 (LAMP), which can direct the expressed antigen to MHC Class II-rich compartment, thus skewing ZIKV antigen presentation towards a strongly Th2-biased response and generation of immune memory; and (2) flash nanocomplexation (FNC)-produced small (~40 nm) ZIKV/LAMP DNA nanoparticles to enable drainage to the dLNs following s.c. injection, thus enhancing the immune response and allowing for substantial reduction in vaccination dose. We will first engineer the DNA nanoparticles with different sizes and narrow distribution using the FNC method, and characterize their stability, surface properties, and ability to transfect DCs; then determine the effect of NP size on LN-draining efficiency and gene expression in DCs in different dLNs; and lastly demonstrate the robust immune responses elicited by LN-targeting NPs with LAMP/ZIKV DNA vaccine, and characterize the subtypes of anti-ZIKV response. This timely study will not only develop a more effective ZIKV vaccine and support further testing of the next generation of Zika vaccine, but also provide a potent DNA vaccine platform against other emerging pathogens.

项目概要该项目的目标是开发能够有效输送至局部引流淋巴的 DNA 纳米粒子通过皮下 (s.c.) 给药促进树突状细胞 (DC) 转染、抗原呈递和 T 细胞在淋巴结中的激活，并引发强大的抗体滴度和免疫学对抗寨卡病毒的记忆。随着巴西寨卡病毒（ZIKV）疫情向南北蔓延美洲、欧洲、亚洲，世界各地的团队都在竞相研发 ZIKV 疫苗。脱氧核糖核酸疫苗在易于生产、优异的稳定性和较长的保质期方面具有许多优点，多价能力和快速的开发周期。第一代候选疫苗是用作为主要免疫原的巴西 ZIKV 毒株的前膜和包膜序列。他们有作为裸质粒 DNA 疫苗通过肌内 (i.m) 注射进行测试，并生成在小鼠和非人灵长类动物中有效的中和抗体。然而，裸DNA疫苗本质上具有产生Th2反应和免疫记忆的效率低，并且由于低浓度而需要更高的DNA剂量注射部位有丰富的抗原呈递细胞 (APC)。本研究将直接解决这些问题通过结合两种新方法来设计更有效的 ZIKV DNA 疫苗来应对挑战：(1) 嵌合体编码 ZIKA 包膜蛋白序列和溶酶体相关膜蛋白 1 的 DNA 构建体 (LAMP)，它可以将表达的抗原引导至富含 MHC II 类的区室，从而使 ZIKV 抗原产生偏差呈现出强烈的 Th2 偏向反应和免疫记忆的产生； (2) 闪光灯纳米络合 (FNC) 产生的小型（约 40 nm）ZIKV/LAMP DNA 纳米颗粒能够引流到 dLN 以下 s.c.注射，从而增强免疫反应并大幅减少疫苗接种剂量。我们将首先设计不同尺寸和窄分布的 DNA 纳米粒子使用FNC方法，表征其稳定性、表面性质和转染DC的能力；然后确定 NP 大小对不同 dLN 中 DC 的 LN 引流效率和基因表达的影响；和最后证明了 LN 靶向 NP 使用 LAMP/ZIKV DNA 疫苗引发的强大免疫反应，并描述抗 ZIKV 反应的亚型。这项及时的研究不仅将开发出更有效的 ZIKV 疫苗并支持下一代寨卡疫苗的进一步测试，同时也提供了有效的 DNA 针对其他新兴病原体的疫苗平台。