Infection Immunology (Infection & Immunity)
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Team der Forschungsgruppe "Infection & Immunity" The "Infection & Immunity" group focuses on research into the host response to infection and vaccination in the context of cohort studies in humans with a focus on SARS-CoV2 infections, the human immunodeficiency virus (HIV), oncogenic human papillomaviruses (HPV), Mycobacterium tuberculosis (MTB) and helminth infections (together with AG Inge Kroidl). This includes the molecular characterization of the infectious pathogen, the identification of correlates of immune protection against infection and disease progression, the discovery of biomarkers for TB diagnostics and therapy monitoring, the characterization of HIV reservoirs and the mapping of B-cell epitopes after vaccination. In addition to the analysis of systemic cellular and humoral immune responses, the group has expertise in the analysis of effector cell subsets in human tissue samples using multiplex fluorescence immunohistochemistry in combination with the detection of viral nucleic acids using in situ hybridization to study the microenvironment of virus-infected cells. The ability to bioinformatically analyze large RNA sequencing data and microscopy images is being further expanded.
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Polychromatic flow cytometry and flow cytometric cell sorting for in-depth characterization of human cells
The laboratory is equipped with a 13-color Beckmann Coulter CytoFLEX flow cytometer and has extensive experience in the flow cytometric analysis of human cells. This includes the characterization of pathogen-specific T cells after in vitro restimulation or by HLA tetramer staining as well as all other cell types in the blood. The laboratory uses the CyTUM-MIH flow cytometry core unit (weblink: http://www.fcfu.mikrobio.med.tum.de/CyTUM-MIH/home) for flow cytometric cell sorting at biosafety level S2 and S3 to enable downstream analysis of DNA and RNA in sorted cells.
Peptide microarray analyses of pathogen-specific antibody reactions
The laboratory serves as the central peptide array laboratory for the European HIV Vaccine Alliance(http://www.ehv-a.eu/). We use a fully validated peptide microarray assay to map and analyze specific antigenic regions recognized by the antibodies elicited by the vaccine. The technology is suitable for both clinical and preclinical samples. The technology enables reactivity testing against up to 8000 different peptides in triplicate in multiple samples simultaneously. The design of the peptide microarray is carried out in collaboration with Dr. Georgios Pollakis (University of Liverpool, weblink https://www.liverpool.ac.uk/infection-and-global-health/staff/georgios-pollakis/publications/). The downstream bioinformatic analysis of the generated data is well established and can be adapted depending on the research question.
Histological characterization of tissue resident cells in their natural microenvironment
Histological methods provide valuable information about the cells in their natural environment, e.g. about their relative position in the tissue and about cell-cell interactions. To exploit this information, the laboratory has introduced multiplex fluorescence immunohistochemistry for numerous cell markers in combination with the detection of nucleic acids by in situ hybridization. Customizable semi-automated analysis tools have been established for the most common staining panels. Via the Core Facility Bioimaging at the Biomedical Center (BMC)(https://www.bioimaging.bmc.med.uni-muenchen.de/index.html), we have access to various microscopes depending on the application.Transcriptome analysis by RNA sequencing
Transcriptome analysis is well established in the laboratory, with bioinformatics workflows developed in-house. A particular focus of our research is the analysis of low abundance cell populations to discover novel biomarkers. We have expertise in high-throughput analysis of whole blood collected in clinical trials and observational studies, as well as RNA sequencing of sorted rare cell populations such as pathogen-specific low-abundance T cells.
Laboratory services, consulting services and international clinical trial logistics
The laboratory can provide laboratory services for the processing and analysis of human samples for clinical trials in Munich and elsewhere. This includes the processing of blood samples (plasma, serum, peripheral blood mononuclear cell isolation). The group also has extensive experience in supporting large international clinical trials with a particular focus on Africa. We therefore offer support in sample and reagent logistics, capacity building and the analysis process in various national and international studies. Please contact us for further information.
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Platform: Polychromatic flow cytometry and flow cytometric cell sorting for in-depth characterization of human cells
Representative dot plots for the phenotypic characterisation of pathogen-specific CD4 T cells. The laboratory is equipped with a 13-colour Beckmann Coulter CytoFLEX Flow Cytometer and has vast experience in the flow cytometric analysis of human cells. This includes characterization of pathogen-specific T cells after in vitro restimulation or by HLA tetramer staining as well as any other cell type in the blood. The laboratory uses the Flow Cytometry core unit CyTUM-MIH for flow cytometric cell sorting at biosafety S2 and S3 level to allow for downstream analyses of DNA and RNA in sorted cells.
Platform: Peptide microarray analyses of pathogen-specific antibody responses
Antibody epitope mapping using peptide microarrays. The laboratory serves as a core peptide array laboratory for the European HIV Vaccine Alliance. We use a fully validated peptide microarray assay to map and analyse specific antigenic regions recognized by antibodies elicited by vaccination. The technology is established for both, clinical and preclinical specimen. The technology allows reactivity testing against up to 8000 different peptides in triplicates in multiple samples simultaneously. Custom design of the peptide microarray is performed in collaboration with Dr. Georgios Pollakis(University of Liverpool). Bioinformatic downstream analysis of generated data is well established and can be adapted depending on the research question.
Platform: Histological characterization of tissue-resident cells in their natural microenvironment
An HIV infected T cell is attacked by a granzyme B expressing T cell. Histological techniques provide valuable information on cells within their natural environment, such as their relative positioning within the tissue and cell-cell interactions. To take advantage of this information, the laboratory has established multiplex fluorescence immunohistochemistry for numerous cell markers in combination with the detection of nucleic acids using in situ hybridization. Customizable semi-automated analysis tools are established for the most common staining panels. We have access to various microscopes via the Core Facility Bioimaging at the Biomedical Center (BMC), depending on the application in question.
Platform: Transcriptome analysis by RNA sequencing
Differential expression of various genes in healthy and sick individuals. Transcriptome analysis is well established within the lab, with bioinformatic workflows developed in house. A particular focus of our research includes the analysis of low abundant cell populations for the discovery of novel biomarkers. We have expertise in high-throughput analysis of whole blood collected in clinical trials and observational studies as well as in low-input RNA sequencing of sorted rare cell populations such as pathogen-specific T cells.
Laboratory services, Consultancies and international clinical study logistics
The laboratory can provide laboratory services for human sample processing and analysis for clinical studies in Munich and elsewhere. This includes the processing of blood samples (Plasma, Serum, Isolation of Peripheral Blood Mononuclear Cells). The group also has tremendous experience in supporting large international clinical with a specific emphasis on Africa. We therefore provide support on sample and reagent logistics, capacity development and the analytic process in multiple national and international study set ups. Please contact us for more information.
