Innate and adaptive defenses against SARS-COV-2 in the oral cavity during acute unvaccinated and breakthrough infections

急性未接种疫苗和突破性感染期间口腔针对 SARS-COV-2 的先天和适应性防御

• 批准号: 10667248
• 负责人: JEFFREY M. JACOBSON
• 金额: $ 48.3万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10667248
• 关键词: 2019-nCoV; Accounting; Acute; Address; Aerosols; Affect; Affinity; Age; Antibody Response; Binding; Biological Assay; Biology; Blood; Brush Cell; CLIA certified; COVID-19; COVID-19 vaccination; COVID-19 vaccine; Cells; Clinical; Clinical Trials; Communicable Diseases; Coughing; Cytology; Data; Data Set; Dyspnea; Enrollment; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Event; Female; Fever; Future; Gender; Gene Expression; Genomics; Goals; Health Professional; Healthcare Systems; Hospitalization; Host Defense; Immune; Immune response; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunologic Factors; Immunologic Markers; Immunology; Immunotherapy; Individual; Infection; Inflammatory Response; Interferons; Invaded; Laboratories; Longitudinal Studies; Measures; Memory; Molecular; Molecular Biology; Monitor; Natural History; Natural Immunity; Oral; Oral Medicine; Oral cavity; Oral mucous membrane structure; Outcome; Participant; Patients; Peripheral; Persons; Prognostic Marker; RNA; RNA vaccine; Recovery; Response Elements; SARS-CoV-2 exposure; SARS-CoV-2 immune response; SARS-CoV-2 infection; SARS-CoV-2 infection history; SARS-CoV-2 positive; SARS-CoV-2 variant; Saliva; Salivary; Sampling; Secretory Immunoglobulin A; Serology; Serum; Severity of illness; Site; Source; Symptoms; Testing; Time; Tongue; Vaccinated; Vaccination; Vaccinee; Vaccines; Variant; Viral; Viral Genome; Viral Load result; Virus; Virus Diseases; Virus Replication; adaptive immune response; anti-viral efficacy; antimicrobial peptide; base; breakthrough infection; cohort; cytokine; digital; field study; genome sequencing; innovation; insight; male; member; mucosal site; multidisciplinary; nasopharyngeal swab; novel; novel diagnostics; oral cavity epithelium; particle; pathogen; post SARS-CoV-2 infection; rapid test; recruit; response; saliva sample; sex; single-cell RNA sequencing; study population; success; tongue sampling; transmission process; unvaccinated; vaccination outcome; vaccine efficacy; vaccine trial; viral RNA; viral transmission; virology; whole genome

Background. Vaccines against SARS-CoV-2 (CoV-2) do not prevent infection and subsequent transmission. Strong evidence now implicates the oral cavity as a primary site of acute CoV-2 infection It is therefore incumbent upon us to search for evidence as to why primary mucosal sites, such as the tongue, remain vulnerable to CoV-2 infection despite vaccination. Our goal. Immune responses are both innate, which are not specific to an invading pathogen, and adaptive, which are specific antibody responses. We hypothesize that interrogating oral epithelial cells and saliva collected from CoV-2 acutely infected unvaccinated and breakthrough-infected subjects will reveal early and specific innate and adaptive immune responses that vary with viral load, viral variants, viral clearance, and patient sex and age. We will collect samples at two time points from acutely infected unvaccinated participants; one within 7 days and the second within 14 days of testing positive. We will evaluate the relationship among measures of oral mucosal viral replication, innate and adaptive factors from salivary, cytological, and serological sources and determine their associations with quantitative viral measures in the oral mucosa. Innate factors to be analyzed in the saliva include, but not limited to, cytokines, antimicrobial peptides, interferons (IFN) and secretory IgA that are correlated with early infection events. To cross validate our results we will also characterize viral strains and develop novel PCR-based assays for intracellular CoV-2 RNA. Studies will include single cell RNA sequencing to analyze virus-induced gene expression changes in infected and bystander cells from the tongue. Full genome sequencing will be used to determine if viral variants are present and correlate oral immune profiles with variants. Finally we will develop digital PCR assays to selectively amplify sub genomic CoV-2 RNA as a rapid measure of infected cells. Once this baseline information is established, we will compare the adaptive and innate oral immune factors of CoV-2 vaccinated and breakthrough study participants, respectively, to the oral immune responses in natural infection. How will we advance the field? This study will allow us to chronicle for the first time the impact of vaccination on oral immune responses of healthy individuals and compare them to immune responses from COVID-19-exposed individuals. We anticipate that our integrated accounting of the oral immune responses during the natural history of SARS-CoV-2 infections will provide an invaluable reference data set for patients, healthcare professionals and those measuring the efficacy of antiviral treatments, immunotherapies and vaccines.

背景。针对SARS-COV-2（COV-2）的疫苗不会防止感染，随后 传播。现在，有力的证据表明口腔是急性COV-2的主要部位 因此，感染因此有责任寻找有关主要粘膜的证据 尽管疫苗接种，例如舌头等地点仍然容易受到COV-2感染的影响。 我们的目标。免疫反应都是先天的，不是针对入侵病原体的特定的， 和自适应，这是特定的抗体反应。我们假设询问口头 从COV-2收集的上皮细胞和唾液急性感染未接种疫苗和 突破感染的受试者将揭示早期和特定的先天和适应性免疫 因病毒载荷，病毒变异，病毒清除以及患者性别和年龄而变化的反应。 我们将从急性感染未接种疫苗的参与者的两个时间点收集样品；一 在7天内，第二次在测试阳性后的14天内。我们将评估关系 在口腔粘膜病毒复制的措施中，唾液的先天和适应性因素 细胞学和血清学来源，并确定其与定量病毒的关联 口腔粘膜的措施。唾液中要分析的先天因素包括但不限 与与之相关的细胞因子，抗菌肽，干扰素（IFN）和分泌IgA 早期感染事件。跨越验证我们的结果，我们还将表征病毒株和 开发用于细胞内COV-2 RNA的新型基于PCR的测定。研究将包括单细胞 RNA测序分析感染和旁观者中病毒诱导的基因表达变化 细胞从舌头上。完整的基因组测序将用于确定病毒变体是否为 目前并将口服免疫谱与变体相关联。最后，我们将开发数字PCR 选择性扩增基因组COV-2 RNA作为对感染细胞的快速度量的测定。一次 建立了此基线信息，我们将比较适应性和先天的口腔免疫 COV-2分别接种和突破性的研究参与者的因素为口腔免疫 自然感染的反应。 我们将如何发展该领域？这项研究将使我们能够首次记载影响 对健康个体的口服免疫反应的疫苗接种，并将其与免疫进行比较 来自共同19岁的个体的反应。我们预计我们的综合会计 SARS-COV-2感染的自然病史期间的口腔免疫反应将提供 针对患者，医疗保健专业人员和测量的患者的参考数据集 抗病毒治疗，免疫疗法和疫苗的功效。