SARS-CoV2 Immunity and Cell Biology

SARS-CoV2 免疫和细胞生物学

• 批准号: 10927964
• 负责人: Jonathan W Yewdell
• 金额: $ 30.15万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10927964
• 关键词: 2019-nCoV; A549; ACE2; Affinity; Alanine; Antibody Specificity; Binding; Biological; Biological Assay; Cells; Cellular biology; Coronavirus; Coronavirus Infections; Epitopes; Goals; Histocompatibility Antigens Class I; Human; Interferometry; Libraries; MHC Class I Genes; Monoclonal Antibodies; Mus; Nucleocapsid Proteins; Pathway interactions; Peptide Hydrolases; Process; Proteins; Proteome; Recombinants; Reporter; SARS-CoV-2 immunity; SARS-CoV-2 spike protein; Sleeping Beauty; Stomatitis; System; TMPRSS2 gene; Transfection; Transposase; Vesicular stomatitis Indiana virus; Viral Physiology; Virus; Virus Receptors; antigen processing; chemokine; cytokine; peptide I; receptor; seasonal coronavirus; translatome; vector

We have generated Sleeping Beauty transposase vectors that drive expression of the SARS-CoV-2 receptor ACE2 and or protease TMPRSS2 in host cells and use these vectors to create permanent transfected lines of A549, 293FT, and BHK-21 cells. We have generated a recombinant vesicular stomatitis virus that expresses a reporter fluorescent protein and SARS-CoV-2 spike protein in place of the normal viral receptor. This virus allows us to perform highly accurate neutralization assays using anti-spike mAbs or polyclonal sera. We have generated several hundred mouse mAbs specific for SARS-CoV-2 spike protein and are in the process of defining their epitopes and biological activity. For this effort, we are generating library in which each residue in spike is replaced with alanine. The library will be used express spike via transfection or by an equivalent recombinant vesicular stomatitis library. We are generating mAbs specific for SARS-CoV-2 nucleocapsid protein. We have characterized the binding of dozens of chemokines to a number of SARS-CoV-2 proteins and have discovered a number of chemokines that bind with high affinity as determined by bio-layer interferometry. We have created a system for generating recombinant seasonal CoVs or recombinant SARS-CoV-2. We are creating seasonal CoVs that express SARS-CoV-2 spike in place of native spike to enable a more realistic neutralization assay. We are also generating recombinant CoVs that express MHC class I peptides to enable studying the effects of CoV infection on the MHC class I antigen processing pathway.

我们已经产生了睡美人转座酶向量，这些媒介驱动宿主细胞中SARS-COV-2受体ACE2和蛋白酶TMPRSS2的表达，并使用这些向量创建了A549、293ft和BHK-21细胞的永久转染线。 我们已经产生了一种重组囊泡性口腔炎病毒，该病毒表达了报告基因荧光蛋白和SARS-COV-2尖峰蛋白，以代替正常病毒受体。 该病毒使我们能够使用抗尖峰MABS或多克隆血清进行高度准确的中和测定。 我们已经产生了针对SARS-COV-2峰值蛋白的数百个小鼠mAb，并且正在定义其表位和生物活性。 为了进行这项工作，我们正在生成库，其中尖峰中的每个残留物被丙氨酸代替。 该库将通过转染或等效的重组囊炎图书馆使用明确的尖峰。 我们正在生成针对SARS-COV-2 Nucleocapsid蛋白的MAB。 我们已经表征了数十种趋化因子与许多SARS-COV-2蛋白的结合，并发现了许多与生物层干扰法确定的高亲和力结合的趋化因子。 我们创建了一个用于生成重组季节性COV或重组SARS-COV-2的系统。 我们正在创建季节性的COV，以代替本地尖峰表达SARS-COV-2尖峰，以实现更现实的中和测定。 我们还生成了表达MHC I类肽的重组COV，以使研究COV感染对MHC I类抗原加工途径的影响。

项目成果

Individuals cannot rely on COVID-19 herd immunity: Durable immunity to viral disease is limited to viruses with obligate viremic spread.

• DOI: 10.1371/journal.ppat.1009509
• 发表时间: 2021-04
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Yewdell JW

Enhanced virulence and waning vaccine-elicited antibodies account for breakthrough infections caused by SARS-CoV-2 delta and beyond.

• DOI: 10.1016/j.isci.2022.105507
• 发表时间: 2022-12-22
• 期刊: ISCIENCE
• 影响因子: 5.8
• 作者: Kwon, Hyung-Joon;Kosikova, Martina;Tang, Weichun;Ortega-Rodriguez, Uriel;Radvak, Peter;Xiang, Ruoxuan;Mercer, Kelly E.;Muskhelishvili, Levan;Davis, Kelly;Ward, Jerrold M.;Kosik, Ivan;Holly, Jaroslav;Kang, Insung;Yewdell, Jonathan W.;Plant, Ewan P.;Chen, Wilbur H.;Shriver, Mallory C.;Barnes, Robin S.;Pasetti, Marcela F.;Zhou, Bin;Wentworth, David E.;Xe, Hang
• 通讯作者: Xe, Hang

Cell surface SARS-CoV-2 nucleocapsid protein modulates innate and adaptive immunity.

• DOI: 10.1126/sciadv.abp9770
• 发表时间: 2022-08-05
• 期刊: Science advances
• 影响因子: 13.6