Project title: "Immune networks of vaccination during pregnancy and beyond"

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Project Title: "Respiratory virus infections and lung immunity"

Please download the project here .

For more information regarding Prof. Gabriel's working group, please click here .

Organ- and age-specific host defense responses in adenovirus-infections cells

Project leader: Madeleine J. Bunders

Organ- and age-specific dynamics and host defense responses in adenovirus-infected epithelial cells

Project leader: Madeleine J. Bunders

Affiliation: III. Department of Medicine, UKE

Background and preliminary data:

Human adenovirus (HAdV) infections are an important cause of respiratory, hepatic and gastrointestinal infections. Especially children and immune-compromised adults can suffer from severe adenovirus infection and reactivation. HAdV comprise of a large family of DNA viruses of which most express proteins that exhibit exceptional capacities to evade immune control of infected cells and establish latent infection, such as E1 and E3 proteins. Until now most studies investigating HAdV infection are performed in cell lines, however our recent findings, using human tissue-based organoid systems, demonstrate that cell lines poorly model HAdV infection in human cells and HAdV-mediated regulation of host defense responses. Using intestinal organoid systems we were able to identify a new target (HLA-F) identifying HAdV-infected epithelial cells to NK cells, which provided children following hematopoietic stem cell transplantion protection by KIR3DS1+ NK cells against severe HAdV reactivation. However, the HAdV strains infect multiple organs resulting in organ-specific infection and tissue damage. To be able to investigate organ-specific HAdV infection, we have extended the HAdV organoids models to study HAdV infection in airway, liver and intestinal organoid systems. Our preliminary studies of the different organ systems indicate that age and organ-specific dynamics of HAdV infection exist and HAdV proteins show organ-specific modulation of host defense mechanisms relating to specific populations at risk. In this project we will investigate organ-specific and age-specific HAdV infection to understand organ-specific HAdV disease.

Hypothesis:

HAdV mediates organ and age-specific disease in humans

Aims and Work Programme:

• To assess the correlation between organ and age specificity of HAdV infectivity and viral production.
• To identify organ-specific and shared pathways in epithelial cells affected by HAdV that mediate viral control in children and adults.

In Aim #1, organoids from gut, lung and liver tissue are generated from infant and adult donors. Organoids will be infected with a specific serotypes including serotypes with specific tropism to an organ (e.g. HAdV40/41) to serotypes with a capacity to infect a broader range of organs (e.g. HAdV5). Infection will be quantified by flow cytometry and microscopy (viral proteins) and viral DNA in a time course. Cellular characteristics of HAdV-infected cells in organoid systems will be determined by flow cytometry and microscopy. Data will be analyzed using regression models to assess organ and age-specific risk factors for viral infectivity and production. To validate these in vitro studies risk factors for HAdV infection will next be tested using data from large cohorts of individuals undergoing HSCT correlating viral load and organ specific symptoms. Together, these tissue-based and cohort based studies will identify characteristics of patients and epithelial cells susceptible for specific-HAdV serotypes in human organs and HAdV dynamics during infection.

In Aim #2, the generation of robust HAdV infection organoid systems now allows to further determine organ- and age-specific pathways affected by HAdV. To this end, scRNAseq analyses of organoids systems have been established and will be employed to 3 organ organoid systems generated from infants and adults with 3 different serotypes. Computational tools have been developed to perform transcriptomics analyses of HAdV-infected cells and compare to HAdV (RNA) negative cells from the same organoid culture. Upon identification of specific pathways these will be validated using blocking antibodies or gene-edited organoids using Crispr/Cas9. Together these studies will identify pathways shared between organs that are affected by specific HAdV strains as well as organ specific pathways that are associated with HAdV disease in children and adults.

Project-related publications: (max. 5) * shared

1. Möller KJ*, Wegner L*, Malsy J*, et int, Bunders MJ. Expanded ILC2s in human infant intestines promote tissue-growth. Mucosal Immunol. 2023; doi: 10.1016/j.mucimm.2023.04.004. Online ahead of print.

2. Jordan-Paiz A*, Martrus G*, Fenja Steinert*, et int, Bunders MJ. CXCR5+PD-1++ CD4+ T cells colonize infant intestines early in life and promote B cell maturation. Cell Mol Immunol. 2023; 20(2):201-213. doi: 10.1038/s41423-022-00944-4.

3. Jung JM, Ching W, et int, Altfeld M*, Belderbos M*, Dobner T*, Bunders MJ*. KIR3DS1 directs NK cell-mediated protection against human adenovirus infections. Sci Immunol. 2021; 6(63):eabe2942. doi: 10.1126/sciimmunol.abe2942.

4. Sagebiel AF, Steinert F, et int, Bunders MJ. Tissue-resident Eomes(+) NK cells are the major innate lymphoid cell population in human infant intestine. Nat Commun.2019; 10(1):975. doi: 10.1038/s41467-018-08267-7.

5. Schreurs RRCE, Baumdick ME*, Sagebiel AF*, et int, Bunders MJ. Human fetal TNF-α-cytokine-producing CD4(+) effector memory T cells promote intestinal development and mediate inflammation early in life. Immunity.2019; 50(2):462-476.e8. doi: 10.1016/j.immuni.2018.12.010.