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.

背景资料。SARS-CoV-2(CoV-2)疫苗不能预防感染和随后的 变速箱。目前有强有力的证据表明口腔是急性CoV-2的主要感染部位 因此，我们有责任寻找证据，说明为什么初级粘膜 尽管接种了疫苗，但舌头等部位仍然容易受到CoV-2感染。 我们的目标。免疫反应既是先天的，也不是针对入侵的病原体的， 和适应性，这是特异性的抗体反应。我们假设审问口供 从急性感染的未接种和未接种的CoV-2收集的上皮细胞和唾液 突破性感染的受试者将揭示早期和特定的先天性和获得性免疫 反应因病毒载量、病毒变异、病毒清除以及患者性别和年龄而异。 我们将在两个时间点从急性感染的未接种疫苗的参与者那里收集样本；一个 在7天内和第二次在14天内检测呈阳性。我们将评估这一关系 在口腔粘膜病毒复制的指标中，来自唾液的先天和适应性因素， 细胞学和血清学来源，并确定它们与定量病毒的关系 在口腔粘膜中采取措施。唾液中需要分析的先天因素包括但不限于 TO、细胞因子、抗菌肽、干扰素和分泌型IgA 早期感染事件。为了交叉验证我们的结果，我们还将对病毒株和 建立新的基于聚合酶链式反应的细胞内冠状病毒2RNA检测方法。研究将包括单细胞 RNA测序分析病毒感染和旁观者基因表达的变化 舌头上的细胞。全基因组测序将被用来确定病毒变体是否 提供口服免疫图谱并将其与变种相关联。最后，我们将开发数字聚合酶链式反应 选择性扩增亚基因组CoV-2 RNA作为感染细胞的快速检测方法。一次 这一基线信息建立后，我们将比较获得性和先天口服免疫 CoV-2疫苗接种和突破性研究参与者对口服免疫的影响因素 对自然感染的反应。 我们将如何推进这一领域？这项研究将使我们能够第一次记录 疫苗接种对健康人口腔免疫反应的影响及其与免疫的比较 新冠肺炎用户的回应。我们预计我们的综合账目将 SARS-CoV-2感染自然过程中的口服免疫反应将提供 为患者、医疗保健专业人员和那些测量 抗病毒治疗、免疫治疗和疫苗的疗效。