Molecular Microbiology (Virology)
Institute of Medical Microbiology and Hygiene
University Regensburg

HIVacToGC

Funding: The Federal Ministry of Education and Research (BMBF) Title: HIV Vaccine Targeting via DNA Origami Nanoparticles to lymph nodes to promote Germinal Center formation

HIV has so far defied all attempts to develop an effective vaccine against it. Nine efficacy trials testing several vaccine candidates have been conducted in the last 25 years, with only one of them resulting in very modest, insufficient protection. One trial, PrEPVacc, is still ongoing.

Yet, in the 40 years since HIV’s discovery, a lot has been learned about the virus, the reaction of the human immune system, and the means how HIV escapes the immune response.
Project partners: Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg – Project co-ordinator, Pharmaceutical Technology, University of Regensburg, Technical University of Munich (TUM), Helmholtz Zentrum München, Tilibit nanosystems GmbH, 2bind GmbH Further information: HIVacToGC Project site Pressemitteilung

NanoNeutVir

Funding: Bundesministerium für Bildung und Forschung (BMBF)

Neutralizing antibodies provide suitable protection against either infection with SARS-CoV-2 virus or at least protection against severe illness e.g. corona virus induced disease 19 (COVID19) and death. To achieve the desired project aims, different competences are required. Therefore, the project brings together experts from the fields of analytical chemistry, molecular immunology, and virology collaboration with biotech industry to generate a product ready to enter the market. Moreover, this platform can be used in the future to provide a chassis for test assays to detected potent immunity against new emerging virus that might come.
Within the NanoNeutVir consortium, the Wagner Lab is involved in the evolution of those components that provide the liposomes specificity, especially to provide recombinant proteins of RBD variants and ACE2. Moreover, the team will test the efficacy of the established assay with qualified patient sera and benchmark the liposome-based assay against conventional neutralization assays.
Project partners: University of Regensburg, Institute for Analytical Chemistry – project coordinator University of Regensburg, Institute for Medical Microbiology and Hygiene, Molecular Microbiology (Virology) University of Marburg Microcoat Biotechnology GmbH, Bernried Further information: https://www.microcoat.de

EHVA

Funding: European Union’s Horizon 2020 Research and Innovation Programme

The European HIV Vaccine Alliance (EHVA) – funded by the European Union’s H2020 Program – aims at discovering and evaluating new prophylactic and therapeutic HIV-1 vaccine candidates. The program is coordinated by Prof. Yves Lévy (INSERM, Paris) and Prof. Giuseppe Pantaleo (CHUV, Lausanne) and comprises 39 partners from 15 countries. Within the alliance, the University of Regensburg coordinates the Vaccine-Discovery- Workpackage for novel antigen candidates, delivery systems and formulations.
The antigens will be based on HIV’s envelope protein (Env, see picture) that is the target for protective antibody responses. A novel generation of Env-variants characterized by improved trimer-stability, immuno-focussing towards target sites of broadly neutralizing antibodies, and immuno-silencing of non-neutralizing sites will be generated by rational design and evolutionary approaches (e.g. selection from libraries of Env-variants). The generated variants will be compared with the best Env-antigen currently available (BG505.SOSIP) by means of biochemical, biophysical and immunological tests.
Moreover, novel delivery systems, including new types of virus-like particles (VLPs), and formulations will be assessed for their modulatory capacity on the strength, breadth and quality of the immune responses elicited by the novel Env antigens. In addition, our team will develop a framework to design novel DNA-vaccine vectors that elicit high-quality immune responses matching pre-defined criteria that can, for instance, be derived from the knowledge about correlates of protection. The new DNA vectors, as well as novel approaches based on RNA delivery, novel poxviruses or VSV-based viruses will be benchmarked against the PTVDC-DNA-vaccine that has previously been developed and clinically tested by our group. The best-in-class antigens and delivery systems will then be assessed in non-human primate studies for safety and immunogenicity to identify the candidates that will be furthered into clinical development. Finally, our team also closely cooperates with the partners in the immune profiling platform, the data management / integration / down-selection platform, and the clinical trials platform.
Further information: Project website: http://www.ehv-a.eu/ FactSheet: http://www.ehv-a.eu/EHVApdf.pdf EC project webpage: http://ec.europa.eu/programmes/horizon2020/en/news/european-alliance-accelerate-new-concepts-hiv-vaccine-research

HFVac3

Funding: Innovate UK Title: Clinical trial of a DIOS Trivalent Haemorrhagic Fever Vaccine

The HFVac3-project is based on the predecessor projects EVAC and Tri-LEMvac that also were funded by Innovate UK. Within the EVAC project, a streamlined process for vaccine generation starting with in silico design of antigen sequences, over the generation of delivery vectors (DNA, adenovirus, vaccinia virus) and rapid testing for immunogenicity in small animal models (mice and guinea pigs) – all steps to be completed within one year – had been developed.
In a parallel track within the Tri-LEMvac project, proof-of-concept for immunogenicity and (in part) efficacy of such vaccine candidates against three different haemorrhagic fever viruses (Lassa, Ebola, and Marburg virus) has been obtained. Now, the outcomes of these two projects will be joined to come up with a clinical vaccine candidate and start a phase I clinical trial to assess the safety and immunogenicity of this vaccine candidate addressing the three viral diseases.The Wagner lab contribution to the project encompasses the generation and characterization of the MVA vaccine candidate vector suitable for GMP manufacturing.
Project partners: Prof. Jonathan Heeney (University of Cambridge) – project co-ordinator, Prof. Ralf Wagner (University of Regensburg, Institute for Medical Microbiology and Hygiene, Molecular Microbiology (Virology), DIOSynVax Ltd.

MR-BioQuant

Funding: BMBF VIP+ Full title: Ultrasensitive Magnetresonanz für die in vitro Biomarkerquantifizierung in Diagnostik und Therapie Disease-related proteins and cells play a major role in biomedical research and clinical care. Although the quantitative detection of these biomarkers is becoming increasingly important, there are deficits due to the large variability and the low comparability of the results. The BMBF-funded project MR-BioQuant is intended to set the stage for the use of an innovative technology for precise and reproducible biomarker quantification with a sensitivity in the nanomolar concentration range.
The physical basis of the technology is the reversible binding of hyperpolarized xenon to a sensor biomolecule that recognizes the biomarker of interest, but can also be detected indirectly via the xenon NMR signal with a so-called saturation transfer. The particular potential of this measurement method developed at PTB (Physikalisch-Technische Bundesanstalt) is that it can deliver correct results on biological samples irrespective of variations in the sample composition or change of the measurement platform. Exemplary biosensors are to be generated by enabling antibodies to bind xenon. With the help of these sensor molecules, selected biomarkers should be quantified using the BioQuant method. The potential superiority of the approach is to be validated by benchmarking with conventional methods. The method’s high innovative potential, especially for biomedical research tasks, will be demonstrated in applications from vaccine development. These application examples, together with the implementation of the measurement method on different device platforms, should form the basis for an economic utilization of the BioQuant method by life science research companies and NMR device manufacturers. Within the MR-BioQuant project, the Wagner Lab is responsible for the generation of suitable biosensor molecules derived from different viruses (recombinant viral proteins) or antibodies targeting viral proteins that enable the validation of the BioQuant method.
Project partners: Physikalisch-Technische Bundesanstalt Braunschweig und Berlin (PTB) – Coordinator Universität Regensburg, Institut für Mikrobiologie und Hygiene, Molekulare Mikrobiologie (Virologie) (UR-MED) Universität Regensburg, Institut für Biophysik und Physikalische Biochemie (UR-BIO)

VIROFIGHT

Funding: EU – Horizon 2020 Full title: European Consortium for the Development of Novel Therapies against Viruses such as SARS-CoV-2, HIV, Influenza or Hepatitis

Current antiviral drugs constitute small molecules that target and inactivate typical proteins or enzymes of selected viruses. Within the EU-funded Virofight project (coordinated by the Technical University of Munich), nanometer sized shells shall be developed that enclose viruses, thus neutralizing them. This new approach might allow targeting of diverse viruses by employing the same method.
Virus infections affect millions of people each year. They cause severe human suffering and enormous costs to society. The current COVID-19 pandemic is only one of many examples. For more than 70 % of viruses listed by the WHO, no therapy is available, and the existing antiviral drugs have to be administered very shortly after infection to be effective. The Virofight consortium proposes a new approach to fight virus infections that shall make it possible to counter the shortage of broadly applicable antivirals and also to target newly emerging pathogens. The aim is to develop prototypes of nano-shells and to principally prove that they are capable of enclosing different viruses. Different technologies will be applied during the development of these biocompatible nano-shells such as DNA origami, protein design and in-vitro-evolution. The interior of the shells will be coated with molecules that are tuned to certain viruses and ensure a strong and specific interaction. These binding effects shall be assessed for different viruses on a laboratory scale. To achieve the desired project aims, different competences are required. Therefore, the project brings together experts from the fields of supermolecular chemistry, molecular nano-engineering and virology. Within the Virofight consortium, the Wagner Lab is involved in the evolution of those components that provide the nano-shells with specificity toward the different viruses. Moreover, the team will test the efficacy of the nano-shell protoypes against HIV, influenza or corona viruses.
Project partners: Technical University Munich (TUM; Germany) – project coordinator Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg (Germany) Aarhus University (Denmark) ARTTIC S.A.S. (France) National Institute of Chemistry (Slovenia) Further information: Virofight Website EU Cordis portal Our press release on VIROFIGHT project

Assembly and functionalization schematic of DNA-Origami nanoshells as published in Monferrer et al, Science physical reports, 2023 https://doi.org/10.1016/j.xcrp.2022.101237

TREATCANCERV

Funding: Eurostars Full title: Treating cancer by targeting endogenous retroviral fossils within the human genome

Human endogenous retroviruses (HERVs) are transposable retroviral elements accounting for about 8 % of the human genome. They are ancient relicts acquired through multiple infections of the germ line by now extinct exogenous retroviruses.
Although many HERV sequences have acquired inactivating mutations during human evolution, some HERVs retain protein-coding functions, which partly play important roles in human physiology and pathology. For example, correlation between HERVs and human cancer has been described for melanoma, breast cancer, germ cell tumours, renal cancer and ovarian cancer where HERV Envelope (Env) proteins are overexpressed compared to healthy tissues. The TREATCANCERV partners therefore aim at developing therapeutic vaccines as well as therapeutic monoclonal antibodies (mAbs) targeting these HERV Env proteins. The Wagner lab contributes to this project by designing antigens based on the Env protein for different HERV strains, which will then be used for the production of hybridoma cell lines expressing monoclonal antibodies targeting these antigens. The project partner Sirion-Biotech will produce adenoviral vectors carrying the same antigens while our project partners from the University of Copenhagen and the Danish company InProTher will develop the therapeutic vaccination approach based on a Virus-Like-Vaccine (VLV).
Project partners: Hervolution ApS (Copenhagen, Denmark) Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg (Germany) University of Copenhagen (Copenhagen, Denmark) Sirion-Biotech (Munich, Germany) Further information: https://hervolutiontx.com https://www.sirion-biotech.com/

DeeP-CMV

Funding: EU – Horizon 2020 Full title: Diagnostic, therapy and prevention before cytomegaloviral infection The human cytomegalovirus (CMV) is one of the most important pathogens in humans and triggers as an opportunistic pathogen in certain high-risk situations, e.g. under drug-induced immunosuppression, after bone marrow or organ transplantation and during pregnancy, it causes severe, sometimes life-threatening infections. CMV infection is the most common infection-related risk for neonatal defects today. Currently, there is no vaccine available and decades of experience with anti-herpesviral therapies have not been able to solve the problems of low tolerability and viral resistance.
The established, reliable diagnostics (genome PCR detection, antigenemia test, and serology) are limited in that they are not predictive of viral load and about the CMV-associated pathogenesis. This limitation is particularly relevant to the major problem of CMV infection during pregnancy about viral transmission to the unborn child and the likelihood of a pathogenic course. In addition, there is an urgent medical need for improved CMV treatment strategies, pharmacologic prophylaxis and test strategies. The cooperating partners are aiming a targeted knowledge gain in diagnostic, therapy and prevention. Therefore, antibody signatures are a target for CMV disease and transmission prediction. Serological and molecular test strategies can improve the early diagnosis and specification of viral load. Specific CDK-inhibitors are designed for a selective inhibition of viral CDK-dependent processes. Finally, for the prevention before infection, the development of a vaccine is in focus. Within the DeeP-CMV project, the Wagner Lab of the University of Regensburg develops in cooperation with Sirion Biotech different adenoviral, MVA and mRNA based vectors. Development shall comprise various antigen designs addressing induction of T- and B-cell responses as well as various vector optimizations. Newly generated vectors are characterized in in vitro and in vivo models. In addition, the development of a RT-qPCR based test is supported for a more detailed and efficient testing compared to conventional methods. The higher sensitivity of RT-qPCR compared to the conventional ELISpot would also make it possible to test new-borns.
Project partners: Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU, Germany) – project coordinator Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg (Germany) Virion/Serion (Würzburg, Germany) Mikrogen GmbH (Germany) Sirion Biotech GmbH (Germany) Lead Discovery Center (LDC, Germany) Further information: Bavarian Research Foundation Sirion Biotech GmbH University Witten/Herdecke

Inhibiting SARS-CoV-2 N-Protein-Mediated Infectivity

The aim of this project is to identify active substances that specifically prevent the packaging of the SARS-CoV-2 genome into the viral particle and thus inhibiting its replication. Packaging of the viral genome into the virus is an essential process in the viral life cycle whereby the nucleocapsid protein (N) specifically binds to the RNA genome and organizes its packaging in the virus. In particular, SARS N undergoes liquid-liquid phase separation (LLPS) with RNA, which contributes to viral transcription, replication and ultimately virion assembly.
In this project, a structure- based and cellular screening system will be established that can identify molecules that interact with N. Moreover, the functional effects of these interactors on RNA-binding and packaging will be analysed. Ultimately, to determine effective therapeutics, their impact on viral replication and infectivity will be directly assessed in cellular systems. For this we already successfully established a stable cell line that can be induced by doxocycline to express SARS-CoV-2 N protein. This cell line is the basis for the establishment of a cell-based screening assay using phase separation as a readout.
Partner: AG Längst, Fraunhoferinstitut (Germany) Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg (Germany)

TiKoCo

Funding: Bavarian State Ministry of Science and Arts (StMWK) and National Research Network of the University Medicine (NUM; applied surveillance and testing; B-FAST) Full title:Tirschenreuth-Kohorte-Covid-19 study
The Bavarian county of Tirschenreuth was one of the first “Corona hot-spot” areas in Germany, as it was hit by a severe outbreak of SARS-CoV-2 connected to a beer-festival in March 2020, which lead to a high incidence of (PCR-positive) cases and a remarkably high case fatality rate (CFR) of 11.5%. As it was unclear at this point of time if the CFR is based on insufficient identification of cases or on other unknown effects, the Tirschenreuth-Kohorte-Covid-19 (TiKoCo19) study was started to determine the actual seroprevalence in the county in collaboration with the FAU Erlangen and the LMU Munich.
The study utilized a population based randomized sampling approach from government register, subsequently, 6608 residents of the Tirschenreuth area aged at least 14 years, were invited to participate in the study. Out of those, 4203 (63.6 %) participated in the study. In the following, blood samples were/will be collected at a central blood drawing at three different time points. The blood samples are analyzed for SARS-CoV-2 antibodies utilizing a variety of tests: the UR inhouse-ELISA, the commercially available Roche COBAS-Elecsys tests and further commercially available tests (e.g. by Shenzhen YHLO Biotech the market leader in Asia). On top, other tests as surrogate and wildtype neutralization assays, binding/neutralization assays regarding the arising variants of concern are used. The aim of the project is to monitor SARS-CoV-2 seroprevalence, the surveillance detection ratio and infection fatality ratio as well as antibody stability, reinfections, vaccine effects and other possible effects/correlations with e.g. age, profession or lifestyle factors. Publications: Wagner R, Peterhoff D, Beileke S, Günther F, Berr M, Einhauser S, Schütz A, Niller HH, Steininger P, Knöll A, Tenbusch M, Maier C, Korn K, Stark KJ, Gessner A, Burkhardt R, Kabesch M, Schedl H, Küchenhoff H, Pfahlberg AB, Heid IM, Gefeller O, Überla K. Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020. Viruses. 2021 Jun 10;13(6):1118. Einhauser S, Peterhoff D, Niller HH, Beileke S, Günther F, Steininger P, Burkhardt R, Heid IM, Pfahlberg AB, Überla K, et al. Spectrum Bias and Individual Strengths of SARS-CoV-2 Serological Tests—A Population-Based Evaluation. Diagnostics. 2021; 11(10):1843. Peterhoff D, Einhauser S, Beileke S, Niller HH, Günther F, Schachtner M, Asbach B, Steininger P, Tenbusch M, Peter AS, Gessner A, Burkhardt R, Heid IM, Wagner R, Überla K. Comparative Immunogenicity of COVID-19 Vaccines in a Population-Based Cohort Study with SARS-CoV-2-Infected and Uninfected Participants. Vaccines (Basel). 2022 Feb 18;10(2):324. Einhauser S, Peterhoff D, Beileke S, Günther F, Niller H-H, Steininger P, Knöll A, Korn K, Berr M, Schütz A, et al. Time Trend in SARS-CoV-2 Seropositivity, Surveillance Detection- and Infection Fatality Ratio until Spring 2021 in the Tirschenreuth County—Results from a Population-Based Longitudinal Study in Germany. Viruses. 2022; 14(6):1168.
Partners: Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg Institute of Clinical Microbiology and Hygiene, University Hospital Regensburg Institute of Clinical and Molecular Virology, University Hospital Erlangen Statistical Consulting Unit StaBLab, Department of Statistics, LMU Munich Department of Genetic Epidemiology, University of Regensburg Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg University Children’s Hospital Regensburg (KUNO) at the Hospital St. Hedwig of the Order of St. John, University of Regensburg Bayerisches Rotes Kreuz, Kreisverband Tirschenreuth Department of Medical Informatics, Biometry and Epidemiology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU)

CoVaCo

Full title: Course of COVID-19 in vaccinated and non-vaccinated people

The COVID-19 vaccines approved in Germany show a high effectiveness against laboratory-confirmed COVID-19 diseases. Although the vaccines are highly effective in the pivotal studies, it can be assumed that there will a significant number of breakthrough infections due to the large number of those who will be vaccinated in a short time and emerging new virus variants.

The focus of the project is the clinical, immunological and virological investigation of SARS-CoV-2 infections in COVID-19 vaccinated people compared to unvaccinated people. For this purpose, a multi-center prospective cohort study in people with newly diagnosed SARS-CoV- 2 infections is planned.
Project partners: University Hospital Erlangen (Germany) – study coordinator Molecular Microbiology (Virology), Institute of Medical Microbiology and Hygiene, University of Regensburg (Germany) University Hospital Augsburg (Germany) Hospital of the University of Munich (LMU; Germany) Klinikum rechts der Isar of the Technical University of Munich (TUM; Germany) University Hospital Würzburg (Germany) Further information: https://www.virologie.uk-erlangen.de/covako/ Our press release on the CoVaCo project