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

Please download the project here .

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.

Immune regulation and impaired vaccine responsiveness in autoimmune liver diseases

Project leader: Prof. Dr. Ansgar W. Lohse and Dr. Johannes Hartl

Affiliation: I. Medizinische Klinik und Poliklinik, UKE

Background and preliminary data:

For patients with autoimmune disease vaccinations against common pathogens are generally recommended, especially when, or prior to, receiving immunosuppressive therapy. However, we do not really know, if patients with autoimmune diseases can mount a normal anti-vaccine response, what influence immunosuppressive therapy has on the anti-vaccine response, and how disease activity and vaccine responsiveness are inter-related(1). For autoimmune liver diseases, in particular for autoimmune hepatitis (AIH), we have some data suggesting there is a relevant impairment of vaccine responsiveness, that this is present both in untreated patients and in patients undergoing immunosuppressive therapy, and that this is distinct from the other major autoimmune liver diseases, primary biliary cholangitis and primary sclerosing cholangitis (2, 3). In fact, it was shown already 30 years ago that patients with newly diagnosed untreated AIH show an almost completely abrogated response to a tetanus toxoid booster vaccination(4). Very recently we could show that response to SARS-CoV2 vaccine was impaired in AIH patients, both in the humoral and the cellular arm of the vaccine response, and that this impairment was present to a similar degree in patients receiving immunosuppressive therapy compared to untreated AIH patients in remission(2).

Hypothesis:

The hypothesis is, that an impairment of the anti-vaccine response is an expression of a generalized autoregulatory immune response of the patients in an attempt to dampen their autoimmune reaction against liver target antigens. It is furthermore hypothesized that this autoregulatory response is largely mediated by interleukin 10 and transforming growth factor beta, two key cytokine found to be elevated in AIH. Finally, it is hypothesized that the characteristic elevation of IgG in AIH is an expression of this autoregulatory response, which is therefore specific for AIH and not to be found in other liver diseases, autoimmune or metabolic.

Aims and work programme:

The aim of the suggested project is to assess this phenomenon of altered anti-vaccine responsiveness more closely looking at different vaccines (influenza, pneumococci, tetanus and SARS-CoV2 – all recommended vaccinations in AIH) deciphering the anti-vaccine response on a molecular level, and in particular to correlate the anti-vaccine response with clinical parameters such as activity of AIH, disease phase (are the patients on the ascending loop of AIH activity, or are they in a descending phase?), and correlate with key cytokine expression. A particular focus will be on IgG-levels, which are thought by some to be an expression of AIH activity(5), while more recent data suggest that raised IgG-levels are a consequence of the immunoregulatory response of the host, mediated largely by IL10 and TGFß, leading to increased production of IgG. The study will then put a particular focus on a group of patients with AIH, in whom immunosuppressive therapy has recently been tapered, as some of these will relapse, while others stay in remission, the two disease stages being associated with distinct immune profiles, whose kinetics will be characterized and related to vaccine responsiveness. Control patients will be PSC, PBC and non-immune liver diseases (NASH).

Publications:

1. Pape S, Snijders R, Gevers TJG, Chazouilleres O, Dalekos GN, Hirschfield GM, et al. Systematic review of response criteria and endpoints in autoimmune hepatitis by the International Autoimmune Hepatitis Group. J Hepatol. 2022;76(4):841-9.

2. Duengelhoef P, Hartl J, Ruther D, Steinmann S, Brehm TT, Weltzsch JP, et al. SARS-CoV-2 vaccination response in patients with autoimmune hepatitis and autoimmune cholestatic liver disease. United European Gastroenterol J. 2022;10(3):319-29.

3. Hartl J, Ruther DF, Duengelhoef PM, Brehm TT, Steinmann S, Weltzsch JP, et al. Analysis of the humoral and cellular response after the third COVID-19 vaccination in patients with autoimmune hepatitis. Liver Int. 2023;43(2):393-400.

4. Lohse AW, Kogel M, Meyer zum Buschenfelde KH. Evidence for spontaneous immunosuppression in autoimmune hepatitis. Hepatology. 1995;22(2):381-8.

5. Hartl J, Miquel R, Zachou K, Wong GW, Asghar A, Pape S, et al. Features and outcome of AIH patients without elevation of IgG. JHEP Rep. 2020;2(3):100094.

Project Title: Parvovirus B19 evolution and pathogenesis

Please download the project here: Project proposal

For more information regarding Prof. Fischer's / Dr. Lütgehetmann's research, please click here:
Prof. Nicole Fischer
Dr. Marc Lütgehetmann