Bloc transmission of viruses and implications for viral dynamics

病毒的块传播及其对病毒动态的影响

• 批准号: 9589749
• 负责人: Nihal Altan-Bonnet
• 金额: $ 167.75万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 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/9589749
• 关键词: Amino Acid Sequence; Astroviridae; Astrovirus; Autophagocytosis; Autophagosome; Behavior; Biological Models; Cell fusion; Cell membrane; Cells; Cellular biology; Clinical; Coronaviridae; Coronavirus; Cytolysis; DNA sequencing; Disease; Endoplasmic Reticulum; Enteral; Feces; Genetic Variation; Goals; Human; Human poliovirus; Hybrids; Immune; Immune response; In Vitro; Individual; Infection; Innate Immune Response; Lead; Link; Literature; Methods; Mucosal Immune Responses; Multivesicular Body; Mus; Norovirus; Oral; Organism; Parvoviridae; Parvovirus; Pathway interactions; Phosphatidylserines; Population; RNA; Rhinovirus; Role; Rotavirus; Route; Sampling; Severe Acute Respiratory Syndrome; Severities; Surface; Symptoms; Tissues; United States National Institutes of Health; Vesicle; Viral; Viral Genome; Viral Proteins; Virulence; Virus; adaptive immune response; in vivo; late endosome; mouse model; novel; particle; symposium; transcriptome sequencing; transmission process; trend; viral transmission; virology; virus envelope; virus genetics

This past year we have made significant discoveries for each of these objectives summarized below. Identify human viruses (in addition to PV, CVB3 and rhinovirus) that traffic by bloc transmission both in vitro and in vivo. (Marianita Santiana, Sourish Ghosh, Brian Ho) We have discovered that two important human enteric viruses, rotavirus and norovirus, both exit cells as populations using vesicles. We have found that rotavirus vesicles are derived from the plasma membrane and norovirus vesicles are derived from multivesicular bodies. Furthermore, we developed methods to look for whether these two viruses can be transmitted in vivo inside vesicles. With help from our collaborators (Kim Green, Wendy Henderson, Linda Saif) we obtained rotavirus and norovirus infected fecal samples and demonstrated that both viruses are present inside phosphatidylserine-enriched vesicles in fecal samples. We also obtained fecal samples from Astrovirus and Parvovirus infected individuals (thorugh our collaborator David Wang) and developed methods to isolate putative astro or parvo virus carrying fecal vesicles. We are currently analyzing the contents of the vesicles pulled down to determine if they carry these viruses. In addition we are pulling down fecal vesicles from unknown samples and doing RNA and DNA sequencing to identify additional viruses that are being trafficked inside vesicles in feces. Determine the mechanisms by which identified viruses are trafficked out of cells as populations both in vitro and in vivo. (Yael Mutsafi Benhalevy, Teegan Dellibovi-Ragheb) We have identified some of the autophagy machinery being exploited for poliovirus exit in secretory autophagosomes as well as a potential viral protein sequence that is being targeted by the host autophagy machinery to capture poliovirus particles. We have also discovered a novel pathway for coronavirus exit from cells. Contrary to literature that states that these viruses ( SARS, MERS etc) leave cells via the secretory pathway, we have found instead that these viruses are secreted from cells by the fusion with the plasma membrane of an endoplasmic reticulum- late endosome hybrid compartment. Determine the impact, in terms of viral yield, virulence and viral genetic diversity, of viral population carrying vesicles in inter-organismal transmission. (Sourish Ghosh, Marianita Santiana) This past year we have developed an in vivo mouse model system to interrogate the role of vesicles in rotavirus transmission. We have shown that vesicles carrying murine rotavirus are stable and can transmit infection through the oral-fecal route (much like in humans). Moreover we have found that this type of transmission, compared to free rotavirus transmission, results in significantly more severe clinical symptoms, earlier clinical symptom onset, longer disease period and much greater transmission to littermates. We have also begun to examine which tissues are preferentially targeted by viruses carrying populations, what their targeting may mean for the clinical severity observed. We will also begin examining the viral genome diversity in an organism that has been infected by populations as opposed to free particles. Determine the differences, if any, in the host immune responses (innate and adaptive) when infected by bloc transmission versus single particle transmission. (Ying-Han Chen, Sourish Ghosh, Yakey Yaffe) We have found profound differences in the innate and adaptive immune response of cells and organisms respectively to infection by bloc transmission. In summary when cells are infected with high multiplicities of viral genomes we find that they can no longer distinguish among entering non-self RNA and self RNA molecules, leading to an overall suppression of the innate immune response. This is a completely unexpected finding and upends much of what we know about innate immune responses and self/non-self RNA recognition. In addition, we find profound differences in the adaptive immune responses, specifically the mucosal immune response. This coming FY we will investigate if the innate immune behavior (suppression) is linked to the mucosal immune behavior (suppression).

在过去的一年里，我们在以下总结的每个目标上都取得了重大发现。 鉴定通过体外和体内阻断传播进行传播的人类病毒（除PV、CVB 3和鼻病毒外）。（Marianita Rumana，Sourish Ghosh，Brian Ho） 我们已经发现两种重要的人类肠道病毒，轮状病毒和诺如病毒，都使用囊泡作为群体离开细胞。我们发现轮状病毒囊泡来源于质膜，诺如病毒囊泡来源于多泡体。此外，我们开发了方法来寻找这两种病毒是否可以在体内囊泡内传播。在我们的合作者（Kim绿色、Wendy亨德森、琳达赛义夫）的帮助下，我们获得了轮状病毒和诺如病毒感染的粪便样本，并证明这两种病毒都存在于粪便样本中富含磷脂酰丝氨酸的囊泡内。 我们还从星状病毒和细小病毒感染的个体中获得粪便样本（通过我们的合作者大卫王），并开发了分离推定的携带粪便囊泡的星状病毒或细小病毒的方法。我们目前正在分析被拉下来的囊泡的内容物，以确定它们是否携带这些病毒。此外，我们正在从未知样本中提取粪便囊泡，并进行RNA和DNA测序，以确定粪便囊泡中被贩运的其他病毒。 确定已识别病毒在体外和体内作为群体从细胞中运输出来的机制。(Yael Mutsafi Benhalevy，Teegan Dellibovi-Ragheb） 我们已经确定了一些自噬机制被利用脊髓灰质炎病毒退出分泌自噬体，以及一个潜在的病毒蛋白质序列，被靶向的主机自噬机制捕获脊髓灰质炎病毒颗粒。 我们还发现了冠状病毒从细胞中退出的新途径。与文献所述的这些病毒（SARS、MERS等）通过分泌途径离开细胞相反，我们发现这些病毒通过与内质网-晚期内体杂交区室的质膜融合而从细胞分泌。 从病毒产量、毒力和病毒遗传多样性方面，确定携带囊泡的病毒种群在生物体间传播中的影响。 （Sourish Ghosh，Marianita Mrsana） 在过去的一年里，我们已经开发了一种体内小鼠模型系统，以询问囊泡在轮状病毒传播中的作用。我们已经证明，携带鼠轮状病毒的囊泡是稳定的，可以通过口-粪途径传播感染（与人类非常相似）。此外，我们已经发现，与自由轮状病毒传播相比，这种类型的传播导致显著更严重的临床症状、更早的临床症状发作、更长的疾病期和更大的对同窝仔的传播。我们还开始研究哪些组织是携带病毒的人群优先靶向的，它们的靶向可能意味着观察到的临床严重性。我们也将开始检查一个生物体中的病毒基因组多样性，该生物体被群体感染而不是被游离颗粒感染。 确定当通过块传播与单颗粒传播感染时，宿主免疫反应（先天性和适应性）的差异（如果有的话）。 （Ying-Han Chen，Sourish Ghosh，Yakey Yaffe） 我们已经发现细胞和生物体对通过阻断传播的感染的先天性和适应性免疫应答存在深刻的差异。总之，当细胞被高多样性的病毒基因组感染时，我们发现它们不再能够区分进入的非自身RNA和自身RNA分子，导致先天免疫应答的总体抑制。这是一个完全出乎意料的发现，颠覆了我们对先天免疫反应和自我/非自我RNA识别的大部分认识。此外，我们发现适应性免疫反应，特别是粘膜免疫反应的深刻差异。本财年我们将研究先天免疫行为（抑制）是否与粘膜免疫行为（抑制）相关。