Characterizing anti HCMV immunity in an infection model of explanted human lungs

外植人肺感染模型中抗 HCMV 免疫的表征

• 批准号: 421450807
• 负责人: Professor Dr. Reinhold Förster
• 金额: --
• 依托单位: Institut für Immunologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/421450807?language=en
• 关键词: Characterizing; anti; HCMV; immunity; infection

Infection with human cytomegalovirus (HCMV) is life threatening in newborns and in immune compromised adults. Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells play an important role in controlling HCMV infections. HCMV belongs to the family of herpesviridae, a class of viruses known for their notorious species specificity. Thus, HCMV does not infect laboratory animals and therefore information is limited about how CTLs and NK cells restrict HCMV-infected cells in vivo in three-dimensional (3D) organs. To overcome these limitations, we developed a 3D organ culture system that is based on precision cut lung slices (PCLS) from HLA-haplotyped human organ donors. PLCS can be cryopreserved and after thawing can be infected with HCMV. Likewise, HCMV-specific CTLs, expanded from HLA-haplotyped donors, can also be cryopreserved and used to study killing of HCMV-infected cells in PCLS applying 2-photon microscopy. With this innovative experimental setup, we will first build a cell and a tissue bank of HLA-matched PCLS and CTLs. PCLS will be used to profoundly characterize infected cells in the lung that are susceptible to different strains of HCMV using spectral flow cytometry, histology and single cell mRNA sequencing. We then plan to characterize killing dynamics and kinetics of various sub-populations of NK cells and HCMV-specific CTLs by 2-photon microscopy. Finally, we aim to study potential benefits of antibody-dependent cytotoxicity (ADCC) in controlling HCMV applying novel antibodies directed against two different proteins of HCMV that are known to inhibit Fc-gamma receptors.

人巨细胞病毒（HCMV）感染对新生儿和免疫功能低下的成人有生命威胁。细胞毒性T淋巴细胞（CTL）和自然杀伤（NK）细胞在控制HCMV感染中起重要作用。HCMV属于疱疹病毒科，这是一类以其臭名昭著的物种特异性而闻名的病毒。因此，HCMV不感染实验室动物，因此关于CTL和NK细胞如何在三维（3D）器官中体内限制HCMV感染的细胞的信息有限。为了克服这些局限性，我们开发了一种3D器官培养系统，该系统基于来自HLA单倍型人类器官供体的精密切割肺切片（PCLS）。PLCS可以冷冻保存，解冻后可以感染HCMV。同样地，从HLA单倍型供体扩增的HCMV特异性CTL也可以冷冻保存并用于研究在PCLS中应用双光子显微镜杀死HCMV感染的细胞。通过这种创新的实验设置，我们将首先建立HLA匹配的PCLS和CTL的细胞和组织库。PCLS将用于使用光谱流式细胞术、组织学和单细胞mRNA测序深刻表征肺中对不同HCMV毒株敏感的感染细胞。然后，我们计划通过双光子显微镜来表征NK细胞和HCMV特异性CTL的各个亚群的杀伤动力学和动力学。最后，我们的目标是研究抗体依赖性细胞毒性（ADCC）在控制HCMV应用新的抗体针对两种不同的蛋白质的HCMV，已知抑制Fc-γ受体的潜在好处。