Laboratory Studies of Human Respiratory Syncytial Virus and Other Pneumoviruses
人类呼吸道合胞病毒和其他肺病毒的实验室研究
基本信息
- 批准号:10272025
- 负责人:
- 金额:$ 125.84万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:AdultAgeAmino Acid SequenceAmino AcidsAnimal ModelAntigen PresentationAttenuatedAttenuated Live Virus VaccineBackBasic ScienceBloodCCR5 geneCD86 geneCardiopulmonaryCellsChildhoodClinical ResearchCodeCodon NucleotidesComplexComputational algorithmCooperative Research and Development AgreementDeletion MutationDendritic CellsDevelopmentDinucleoside PhosphatesDiseaseDown-RegulationElderlyEpitopesEvaluationExhibitsFamily memberGeneticGenomeGlycoproteinsGoalsHamstersHumanHuman GenomeHuman MetapneumovirusHydrophobicityImmuneImmune responseImmunobiologyImmunologicsIn VitroIndividualInfantInfluenza A virusLaboratory StudyLifeLymphoid TissueMessenger RNAMissense MutationMolecular BiologyMolecular GeneticsMorbidity - disease rateMurine pneumonia virusMusMutationMyelogenousNonstructural ProteinNucleoproteinsOpen Reading FramesPeripheralPhosphoproteinsPlayPneumovirusPolymerasePopulationProductionProtein BiosynthesisProteinsRNARNA VirusesReagentRespiratory Syncytial Virus InfectionsRespiratory Syncytial Virus VaccinesRespiratory Tract DiseasesRespiratory syncytial virusRodentRoleSecondary toSerumSurfaceSystemTechniquesTemperatureTissuesTranslationsUmbilical Cord BloodVaccine Clinical TrialVaccinesVariantViralViral GenomeViral ProteinsVirusVirus ReplicationWorkadaptive immune responseattenuationbasecell motilitycytokinedesigngene synthesisgenome-wideglycoprotein Gimmunogenicitymembermortalitymultiple myeloma M Proteinmutantneutralizing antibodypathogenpreclinical studyprogramsreverse geneticsvaccine candidateviral fitness
项目摘要
We previously performed codon-pair deoptimization (CPD) of various open reading frames (ORFs) of RSV. This is done by rearranging codons using computer algorithms and de novo gene synthesis to increase the content of normally-underrepresented codon-pairs without changing amino acid coding or overall codon usage. CPD is still a new technique and is incompletely understood. It typically has the effect of attenuation. This is thought to be due primarily to reduced efficiency of translation, although this is controversial and other mechanisms may also contribute. In previous years, we produced four RSV mutants by CPD of various combinations of ORFs, and showed that these viruses indeed were attenuated and, unexpectedly, were temperature-sensitive. We also evaluated the genetic stability of two of these viruses under restrictive temperatures, identified potential de-attenuating mutations, and used this information to make a promising vaccine candidate bearing a CPD L polymerase ORF that had desirable feature of attenuation, immunogenicity, and genetic stability. CPD RSV strains have been licensed for development to Codagenix, Inc.
While CPD has been used mostly to generate live-attenuated vaccine candidates, the effects of the converse approach of generating codon pair optimized viruses are still largely unknown. To evaluate effects of codon pair optimization (CPO), we subjected various open reading frames (ORFs) in the RSV genome to CPO by increasing the content of codon pairs that are overrepresented in the human genome without changing overall codon usage and amino acid sequences. This has the potential to increase the expression of the encoded protein(s) and antigenic determinants, and could be useful to generate vaccine candidates with increased immunogenicity. Four viruses were made: Max A (with CPO of NS1, NS2, N, P, M, and SH ORFs), Max B (with CPO of G and F), Max L (with CPO of L), and Max FLC (with CPO of all ORFs except M2-1 and M2-2). Because of the possibility of increased viral replication, each CPO virus was attenuated by the inclusion of a codon deletion mutation (del1313) and a missense mutation (I1314L) in the L polymerase. We found that CPO had no effect on multicycle virus replication in vitro, temperature sensitivity, or specific infectivity. Max A and L, which in common had CPO of one or more ORFs of proteins of the polymerase complex, exhibited global increases in viral protein synthesis. Max B (with CPO of the major antigenic determinants of RSV, the G and F glycoproteins) exhibited decreased protein synthesis, and it alone had reduced single-cycle virus replication in vitro. All CPO RSVs exhibited marginal reductions in replication in mice and hamsters. Surprisingly, the CPO RSVs induced lower levels of serum RSV-neutralizing antibodies in hamsters. This reduced immunogenicity might reflect reduced viral replication and possibly also the decrease in CpG and UpA dinucleotides as immune stimulators. Overall, our study describes paradoxical effects of CPO of an RNA virus on viral replication and the adaptive humoral immune response.
Respiratory syncytial virus (RSV) infects and causes disease in infants and reinfects with reduced disease throughout life without significant antigenic change. In contrast, reinfection by influenza A virus (IAV) largely requires antigenic change. The adaptive immune response depends on antigen presentation by dendritic cells (DC), which may be too immature in young infants to induce a fully protective immune response against RSV reinfections. We therefore compared the ability of RSV and IAV to activate primary human cord blood (CB) and adult blood (AB) myeloid DC (mDC). While RSV and IAV infected with similar efficiencies, RSV poorly induced maturation and cytokine production in CB and AB mDC. This difference between RSV and IAV was more profound in CB mDC. While IAV activated CB mDC to some extent, RSV did not induce CB mDC to increase the maturation markers CD38 and CD86 or CCR7, which directs DC migration to lymphatic tissue. Low CCR7 surface expression was associated with high expression of CCR5, which keeps DC in inflamed peripheral tissues. To evaluate a possible inhibition by RSV, we subjected RSV-inoculated AB mDC to secondary IAV inoculation. While RSV-inoculated AB mDC responded to secondary IAV inoculation by efficiently upregulating activation markers and cytokine production, IAV-induced CCR5 downregulation was slightly inhibited in cells exhibiting robust RSV infection. Thus, suboptimal stimulation and weak and mostly reversible inhibition seem to be responsible for inefficient mDC activation by RSV. The inefficient mDC stimulation and immunological immaturity in young infants may contribute to reduced immune responses and incomplete protection against RSV reinfection.
我们先前对RSV的各种开放阅读框架(ORF)进行了密码子对去优化(CPD)。这是通过使用计算机算法和从头基因合成重新排列密码子来完成的,以增加正常代表性不足的密码子对的含量,而不改变氨基酸编码或总体密码子使用。CPD仍然是一项新技术,人们对它的了解还不完全。它通常具有衰减的效果。这被认为主要是由于翻译效率降低,尽管这是有争议的,其他机制也可能起作用。在过去的几年中,我们通过CPD的ORF的各种组合产生了四个RSV突变体,并表明这些病毒确实是减毒的,出乎意料的是,是温度敏感的。我们还评估了这些病毒中的两种在限制性温度下的遗传稳定性,鉴定了潜在的去减毒突变,并使用这些信息来制备具有减毒、免疫原性和遗传稳定性的期望特征的携带CPD L聚合酶ORF的有希望的疫苗候选物。CPD RSV毒株已被许可开发给Codageland,Inc.。
虽然CPD主要用于产生减毒活疫苗候选物,但产生密码子对优化病毒的匡威方法的效果在很大程度上仍然未知。为了评估密码子对优化(CPO)的效果,我们通过增加在人类基因组中过度表达的密码子对的含量,而不改变总体密码子使用和氨基酸序列,将RSV基因组中的各种开放阅读框(ORF)进行CPO。这有可能增加编码的蛋白质和抗原决定簇的表达,并且可用于产生具有增加的免疫原性的疫苗候选物。制作了四种病毒:Max A(具有NS 1、NS 2、N、P、M和SH ORF的CPO)、Max B(具有G和F的CPO)、Max L(具有L的CPO)和Max FLC(具有除M2-1和M2-2之外的所有ORF的CPO)。由于增加病毒复制的可能性,通过在L聚合酶中包含密码子缺失突变(del 1313)和错义突变(I1314 L)来减毒每种CPO病毒。我们发现,CPO在体外,温度敏感性,或特定的感染性对多重冠状病毒复制没有影响。Max A和L共同具有聚合酶复合体蛋白质的一个或多个ORF的CPO,表现出病毒蛋白质合成的整体增加。Max B(含RSV主要抗原决定簇的CPO,即G和F糖蛋白)表现出蛋白质合成减少,并且其单独在体外具有减少的单循环病毒复制。所有CPO RSV在小鼠和仓鼠中的复制均表现出轻微减少。令人惊讶的是,CPO RSV在仓鼠中诱导较低水平的血清RSV中和抗体。这种降低的免疫原性可能反映了病毒复制的减少,也可能反映了作为免疫刺激剂的CpG和UpA二核苷酸的减少。总的来说,我们的研究描述了RNA病毒的CPO对病毒复制和适应性体液免疫应答的矛盾作用。
呼吸道合胞病毒(RSV)感染并导致婴儿疾病,并在一生中再次感染,疾病减少,而没有显著的抗原变化。相比之下,甲型流感病毒(IAV)的再感染在很大程度上需要抗原改变。适应性免疫反应取决于树突状细胞(DC)的抗原提呈,而树突状细胞在年幼的婴儿中可能还太不成熟,无法诱导针对RSV再感染的完全保护性免疫反应。因此,我们比较了RSV和IAV激活原代人脐带血(CB)和成人血(AB)髓样DC(mDC)的能力。虽然RSV和IAV感染具有相似的效率,但RSV在CB和AB mDC中诱导成熟和细胞因子产生较差。RSV和IAV之间的这种差异在CB mDC中更深刻。虽然IAV在一定程度上激活CB mDC,但RSV并不诱导CB mDC增加成熟标志物CD 38和CD 86或CCR 7,其指导DC向淋巴组织迁移。低CCR 7表面表达与CCR 5的高表达相关,CCR 5将DC保持在发炎的外周组织中。为了评估RSV的可能抑制,我们将RSV接种的AB mDC进行第二次IAV接种。虽然RSV接种的AB mDC通过有效上调活化标志物和细胞因子产生来响应二次IAV接种,但IAV诱导的CCR 5下调在表现出强RSV感染的细胞中被轻微抑制。因此,次优刺激和弱且大部分可逆的抑制似乎是RSV无效激活mDC的原因。低效率的mDC刺激和年幼婴儿的免疫不成熟可能导致免疫应答降低和对RSV再感染的不完全保护。
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)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Ursula Buchholz其他文献
Ursula Buchholz的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Ursula Buchholz', 18)}}的其他基金
Paramyxoviruses as Vaccine Vectors Against Highly Pathogenic Viruses
副粘病毒作为高致病性病毒的疫苗载体
- 批准号:
10272101 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Laboratory And Pre-clinical Studies Of Parainfluenza Viruses
副流感病毒的实验室和临床前研究
- 批准号:
10272021 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Clinical Trials of Vaccines for Respiratory Syncytial Virus and Related Viruses
呼吸道合胞病毒及相关病毒疫苗的临床试验
- 批准号:
10272020 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Laboratory And Pre-clinical Studies Of Parainfluenza Viruses
副流感病毒的实验室和临床前研究
- 批准号:
10927726 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Laboratory Studies of Human Respiratory Syncytial Virus and Other Pneumoviruses
人类呼吸道合胞病毒和其他肺病毒的实验室研究
- 批准号:
10692018 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Paramyxoviruses as Vaccine Vectors Against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
副粘病毒作为针对严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 的疫苗载体
- 批准号:
10692252 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Paramyxoviruses as Vaccine Vectors Against Highly Pathogenic Viruses
副粘病毒作为高致病性病毒的疫苗载体
- 批准号:
10927793 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Paramyxoviruses as Vaccine Vectors Against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
副粘病毒作为针对严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 的疫苗载体
- 批准号:
10272294 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Clinical Trials of Vaccines for Respiratory Syncytial Virus and Related Viruses
呼吸道合胞病毒及相关病毒疫苗的临床试验
- 批准号:
10927725 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
Paramyxoviruses as Vaccine Vectors Against Highly Pathogenic Viruses
副粘病毒作为高致病性病毒的疫苗载体
- 批准号:
10692084 - 财政年份:
- 资助金额:
$ 125.84万 - 项目类别:
相似国自然基金
靶向递送一氧化碳调控AGE-RAGE级联反应促进糖尿病创面愈合研究
- 批准号:JCZRQN202500010
- 批准年份:2025
- 资助金额:0.0 万元
- 项目类别:省市级项目
对香豆酸抑制AGE-RAGE-Ang-1通路改善海马血管生成障碍发挥抗阿尔兹海默病作用
- 批准号:2025JJ70209
- 批准年份:2025
- 资助金额:0.0 万元
- 项目类别:省市级项目
AGE-RAGE通路调控慢性胰腺炎纤维化进程的作用及分子机制
- 批准号:
- 批准年份:2024
- 资助金额:0 万元
- 项目类别:面上项目
甜茶抑制AGE-RAGE通路增强突触可塑性改善小鼠抑郁样行为
- 批准号:2023JJ50274
- 批准年份:2023
- 资助金额:0.0 万元
- 项目类别:省市级项目
蒙药额尔敦-乌日勒基础方调控AGE-RAGE信号通路改善术后认知功能障碍研究
- 批准号:
- 批准年份:2022
- 资助金额:33 万元
- 项目类别:地区科学基金项目
补肾健脾祛瘀方调控AGE/RAGE信号通路在再生障碍性贫血骨髓间充质干细胞功能受损的作用与机制研究
- 批准号:
- 批准年份:2022
- 资助金额:52 万元
- 项目类别:面上项目
LncRNA GAS5在2型糖尿病动脉粥样硬化中对AGE-RAGE 信号通路上相关基因的调控作用及机制研究
- 批准号:n/a
- 批准年份:2022
- 资助金额:10.0 万元
- 项目类别:省市级项目
围绕GLP1-Arginine-AGE/RAGE轴构建探针组学方法探索大柴胡汤异病同治的效应机制
- 批准号:81973577
- 批准年份:2019
- 资助金额:55.0 万元
- 项目类别:面上项目
AGE/RAGE通路microRNA编码基因多态性与2型糖尿病并发冠心病的关联研究
- 批准号:81602908
- 批准年份:2016
- 资助金额:18.0 万元
- 项目类别:青年科学基金项目
高血糖激活滑膜AGE-RAGE-PKC轴致骨关节炎易感的机制研究
- 批准号:81501928
- 批准年份:2015
- 资助金额:18.0 万元
- 项目类别:青年科学基金项目
相似海外基金
PROTEMO: Emotional Dynamics Of Protective Policies In An Age Of Insecurity
PROTEMO:不安全时代保护政策的情绪动态
- 批准号:
10108433 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
EU-Funded
The role of dietary and blood proteins in the prevention and development of major age-related diseases
膳食和血液蛋白在预防和发展主要与年龄相关的疾病中的作用
- 批准号:
MR/X032809/1 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Fellowship
Atomic Anxiety in the New Nuclear Age: How Can Arms Control and Disarmament Reduce the Risk of Nuclear War?
新核时代的原子焦虑:军控与裁军如何降低核战争风险?
- 批准号:
MR/X034690/1 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Fellowship
Collaborative Research: Resolving the LGM ventilation age conundrum: New radiocarbon records from high sedimentation rate sites in the deep western Pacific
合作研究:解决LGM通风年龄难题:西太平洋深部高沉降率地点的新放射性碳记录
- 批准号:
2341426 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Continuing Grant
Collaborative Research: Resolving the LGM ventilation age conundrum: New radiocarbon records from high sedimentation rate sites in the deep western Pacific
合作研究:解决LGM通风年龄难题:西太平洋深部高沉降率地点的新放射性碳记录
- 批准号:
2341424 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Continuing Grant
Doctoral Dissertation Research: Effects of age of acquisition in emerging sign languages
博士论文研究:新兴手语习得年龄的影响
- 批准号:
2335955 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Standard Grant
The economics of (mis)information in the age of social media
社交媒体时代(错误)信息的经济学
- 批准号:
DP240103257 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Discovery Projects
How age & sex impact the transcriptional control of mammalian muscle growth
你多大
- 批准号:
DP240100408 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Discovery Projects
Supporting teachers and teaching in the age of Artificial Intelligence
支持人工智能时代的教师和教学
- 批准号:
DP240100111 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Discovery Projects
Enhancing Wahkohtowin (Kinship beyond the immediate family) Community-based models of care to reach and support Indigenous and racialized women of reproductive age and pregnant women in Canada for the prevention of congenital syphilis
加强 Wahkohtowin(直系亲属以外的亲属关系)以社区为基础的护理模式,以接触和支持加拿大的土著和种族育龄妇女以及孕妇,预防先天梅毒
- 批准号:
502786 - 财政年份:2024
- 资助金额:
$ 125.84万 - 项目类别:
Directed Grant