Design and testing of germline-targeting and boosting immunogens to elicit 10E8-like broadly neutralizing antibodies against HIV
设计和测试种系靶向和增强免疫原,以引发针对 HIV 的 10E8 样广泛中和抗体
基本信息
- 批准号:10188410
- 负责人:
- 金额:$ 90.46万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-07-08 至 2024-06-30
- 项目状态:已结题
- 来源:
- 关键词:ART proteinAdoptive TransferAffinityAnimal ModelAntibodiesAntigensAvidityB cell differentiationB-Cell ActivationB-Cell Antigen ReceptorB-LymphocytesB-cell receptor repertoire sequencingBindingBiological AssayBiophysicsCell SeparationCellsCytometryDataDevelopmentDirected Molecular EvolutionEngineeringEnvironmentEnzyme-Linked Immunosorbent AssayEpitopesExhibitsFlow CytometryFrequenciesFundingGenerationsGenetic StructuresGlycoproteinsGoalsHIVHIV vaccineHIV-1Health PrioritiesHumanImmune responseImmunityImmunizationImmunoglobulin Somatic HypermutationIn VitroIndividualInduced MutationKnock-inKnock-in MouseLeadLiposomesLocationMammalian CellMasksMembraneMembrane ProteinsMessenger RNAMethodsModelingMolecular ConformationMusMutateMutationPassive ImmunizationPathway interactionsPeptidesPhysiologicalPolysaccharidesProtein EngineeringProteinsRNARegimenRepliconResistanceScaffolding ProteinSorting - Cell MovementSpecificityStructureTechniquesTestingVaccinationVaccine ResearchVaccinesVariantViralVirusVirus-like particleWild Type MouseYeastsautoreactivitybasecross reactivitydesignenv Gene Productsexperimental studyglobal healthglycosylationin vivointerestmouse modelnanoparticleneutralizing antibodynext generation sequencingnoveloverexpressionprotein reconstitutionprototyperesponserestraintscaffoldsimian human immunodeficiency virusvaccine candidatevaccine development
项目摘要
Induction of broadly neutralizing antibodies (bnAbs) is a critical unmet goal of HIV vaccine
development. BnAb 10E8 is of particular interest as a vaccine lead due to its (1) very high
breadth, (2) low number of mutations required to develop broad neutralization, (3) low
auto-reactivity, (4) expected high frequency of precursors in the human na'ive B-cell pool and (5)
ability to provide surprisingly potent in vivo sterilizing protection in passive transfer studies.
Key challenges for inducing 10E8-like bnAbs are: (a) the low germline affinity by HIV peptides and
proteins, (b) the severe restriction on bnAb angle of approach imposed by the recessed,
membrane-proximal epitope environment and (c) the absence of the 10E8 epitope from most soluble,
native-like trimers (e.g. SOSIP or NFL or UFO).
A promising strategy to initiate 10E8-like bnAb induction is germline targeting, in which suitable
10E8-class precursors are specifically activated using engineered immunogens, thus selecting BCRs
with the potential to develop broad neutralization in the absence of autoreactivity. This approach
will also help circumvent steric problems associated with the recessed location of the epitope, by
priming precursors with known genetic and structural potential to mature into bnAbs compatible with
MPER steric restraints. In this proposal, we will engineer epitope-scaffold immunogens that
activate 10E8-like precursors, using computational design and directed evolution. We will initially
test immunogens ex vivo using human na'ive B cell sorting. We will further generate knock-in mice
that over-express 10E8-like precursors, and we will use those mice to test B-cell priming and
boosting in vivo, first adoptively transferring knockin B cells to wild- type mice in order to
mimic frequencies of 10E8 bnAb precursor and competitor B cells in humans.
As known bnAbs are highly mutated, vaccine induction of bnAbs following a germline prime will
likely require sequential immunization with other immunogens designed to shepherd affinity
maturation of the B-cell receptor. We will develop different classes of boosting immunogens,
including epitope-scaffolds with more native epitopes, membrane-protein scaffolds and
membrane-bound Env variants stabilized in a conformation to which 10E8 binds strongly. We will
conduct sequential prime/boost immunization experiments in knock-in mice and use ELISA, cytometry,
single B-cell sorting and sequencing and neutralization assays to track and optimize affinity
maturation.
In summary, these studies seek to develop novel HIV vaccine candidates and also to
shift HIV vaccine research towards a reductionist approach based on strategic
identification of bNAb precursors, state-of-the-art protein engineering to develop
germline-targeting and boosting immunogens, development of human Ig knock-in mouse models to enable
testing of human-repertoire-specific vaccines, and in-depth analysis of vaccine-induced affinity
maturation pathways in vivo to guide iterative vaccine optimization.
广泛中和抗体(bnAbs)的诱导是HIV疫苗的一个关键未实现的目标
发展BnAb 10 E8作为疫苗先导特别令人感兴趣,这是由于其(1)非常高的
宽度,(2)发展广泛中和所需的突变数量少,(3)低
自身反应性,(4)人初始B细胞库中预期的高频率前体,和(5)
在被动转移研究中提供令人惊讶的有效体内灭菌保护的能力。
诱导10 E8样bnAb的关键挑战是:(a)HIV肽的低种系亲和力,
蛋白质,(B)由凹进的对bnAb接近角的严重限制,
膜近端表位环境和(c)不存在10 E8表位,
天然样三聚体(例如SOSIP或NFL或UFO)。
启动10 E8样bnAb诱导的一种有前景的策略是种系靶向,其中合适的靶向方法包括:
使用工程免疫原特异性激活10 E8类前体,从而选择BCR
在没有自身反应性的情况下有可能产生广泛中和。这种方法
还有助于避免与表位的凹陷位置相关的空间问题,
引发具有已知遗传和结构潜力的前体成熟为与以下相容的bnAb:
MPER位阻。在这项提案中,我们将设计抗原表位支架免疫原,
使用计算机设计和定向进化激活10 E8样前体。我们将初步
使用人幼稚B细胞分选离体测试免疫原。我们将进一步培育基因敲入小鼠
过表达10 E8样前体的小鼠,我们将用这些小鼠来测试B细胞引发,
体内加强,首先过继转移敲入B细胞至野生型小鼠,
人体中10 E8 bnAb前体和竞争B细胞的模拟频率。
如已知的bnAb是高度突变的,在种系引发后的bnAb的疫苗诱导将使bnAb的免疫原性降低。
可能需要用其他免疫原进行顺序免疫,
B细胞受体的成熟。我们将开发不同种类的增强免疫原,
包括具有更多天然表位的表位支架、膜蛋白支架和
膜结合的Env变体稳定在10 E8强烈结合的构象中。我们将
在基因敲入小鼠中进行顺序初免/加强免疫实验,并使用ELISA,细胞计数,
单个B细胞分选和测序以及中和测定,以跟踪和优化亲和力
成熟
总之,这些研究旨在开发新的HIV候选疫苗,
将艾滋病毒疫苗研究转向基于战略
bNAb前体的鉴定,最先进的蛋白质工程,
生殖系靶向和加强免疫原,开发人IG基因敲入小鼠模型,
测试人类特异性疫苗,并深入分析疫苗诱导的亲和力
成熟途径,以指导迭代疫苗优化。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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WILLIAM R. SCHIEF其他文献
WILLIAM R. SCHIEF的其他文献
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{{ truncateString('WILLIAM R. SCHIEF', 18)}}的其他基金
Design and testing of germline-targeting and boosting immunogens to elicit 10E8-like broadly neutralizing antibodies against HIV
设计和测试种系靶向和增强免疫原,以引发针对 HIV 的 10E8 样广泛中和抗体
- 批准号:
10435499 - 财政年份:2019
- 资助金额:
$ 90.46万 - 项目类别:
Design and testing of germline-targeting and boosting immunogens to elicit 10E8-like broadly neutralizing antibodies against HIV
设计和测试种系靶向和增强免疫原,以引发针对 HIV 的 10E8 样广泛中和抗体
- 批准号:
10655514 - 财政年份:2019
- 资助金额:
$ 90.46万 - 项目类别:
Computational design of novel antigens targeting mature and germline b12
针对成熟和种系 b12 的新型抗原的计算设计
- 批准号:
8463113 - 财政年份:2013
- 资助金额:
$ 90.46万 - 项目类别:
Computational design of novel antigens targeting mature and germline b12
针对成熟和种系 b12 的新型抗原的计算设计
- 批准号:
8117981 - 财政年份:2011
- 资助金额:
$ 90.46万 - 项目类别:
ELASTICITY OF KINESIN UNDER ROTARY AND LINEAR FORCES
旋转力和线性力下驱动蛋白的弹性
- 批准号:
6374822 - 财政年份:2001
- 资助金额:
$ 90.46万 - 项目类别:
ELASTICITY OF KINESIN UNDER ROTARY AND LINEAR FORCES
旋转力和线性力下驱动蛋白的弹性
- 批准号:
6171791 - 财政年份:2000
- 资助金额:
$ 90.46万 - 项目类别:
ELASTICITY OF KINESIN UNDER ROTARY AND LINEAR FORCES
旋转力和线性力下驱动蛋白的弹性
- 批准号:
2865210 - 财政年份:1999
- 资助金额:
$ 90.46万 - 项目类别:
Prediction and Perturbation of Epitopes by Modeling and Immune Responses
通过建模和免疫反应预测和扰动表位
- 批准号:
8897074 - 财政年份:
- 资助金额:
$ 90.46万 - 项目类别:
Computational design of novel antigens targeting mature and germline b12
针对成熟和种系 b12 的新型抗原的计算设计
- 批准号:
8841294 - 财政年份:
- 资助金额:
$ 90.46万 - 项目类别:
Computational design of novel antigens targeting mature and germline b12
针对成熟和种系 b12 的新型抗原的计算设计
- 批准号:
8662174 - 财政年份:
- 资助金额:
$ 90.46万 - 项目类别:
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