18 research outputs found

    Exploring Humoral Immune Responses by Mass Spectrometry: Resolving Structures, Interactions, and Clonal Repertoires of Antibodies

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    In his thesis “Exploring Humoral Immune Responses by Mass Spectrometry”, Maurits den Boer uses mass spectrometry to shed new light on antibody responses. Antibodies play a crucial role in the immune protection against threats like bacteria, viruses, and cancers. When valuable antibodies are discovered, they can therefore be reproduced for use as a medicine. A better understanding of their structures, interactions, and repertoires is therefore key to finding novel treatments for many diseases. In the first part of his thesis, Maurits and coworkers used mass spectrometry to study antibody structures and interactions, leading to two major findings. They first uncovered a mechanism by which Staphylococcus aureus bacteria can evade antibody responses, and how this mechanism may be circumvented in future therapies. Second, he redefined the textbook structure of circulating IgM antibodies by showing that they are universally attached to an extra protein. This may have major implications for how these antibodies function, and their use as therapeutics. In a second line of research, Maurits focused on the development of innovative techniques for antibody repertoire analysis and discovery. Together with coworkers, he explored the use of electron-based fragmentation mass spectrometry, developing methods to obtain valuable pieces of antibody sequence information. Finally, he combined multiple layers of mass spectrometry analysis to discover and fully determine the sequence of a malignant patient antibody. Combined, this demonstrates the promise of mass spectrometry as a compelling new approach for therapeutic antibody discovery

    Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

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    The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers

    Plasticity at the T cell receptor-peptide-major histocompatibility complex class I interface

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    Background – The interaction between the T cell receptor (TCR) on the surface of αβ T cells and the peptide-major histocompatibility complex (pMHC) on the surface of target cells helps αβ T cells to defend the host from virtually any foreign pathogen. To achieve such extensive coverage necessitates substantial T cell crossreactivity. While essential for providing complete immune coverage, T cell crossreactivity can also have negative consequences at it is believed to contribute to a large number of autoimmune diseases an can have fatal consequences when T cells are used therapeutically. The therapeutic deployment of the TCR-pMHC interaction therefore requires thorough understanding of the biochemical characteristics that underpin the interaction. Results – I began my studies by defining the crossreactive profile of an autoreactive T cell clone. Using combinatorial peptide library (CPL) screening, I showed that the HLA-A*0201- restricted, insulin-reactive InsB4 TCR, derived from a patient with type 1 diabetes (T1D) could be strongly activated by peptides arising from three common human pathogens. I further showed that EBV-insulin crossreactivity could be found in some T1D patients but not in healthy controls. The potential for virus-specific T cells to recognise self-antigens in autoimmune patients highlights the importance of understanding the underlying driving features of TCR plasticity. To this end, I set about investigating how two biochemical characteristics can influence the TCR-pMHC interaction, TCR sequence and pMHC flexibility. To explore how TCR sequence impacts TCR function, I profiled the crossreactive nature of an HLA-A*0201-restricted, HIV-1-specific TCR and compared its peptide recognition repertoire to mutant versions of the same TCR with CDR3 amino acid substitutions. These studies inadvertently resulted in the discovery that this TCR could also respond a self-antigen expressed on many HLA-A*0201+ cancer cell lines. Recognition of cancer cells, but not the cognate HIV-derived peptide, could be removed by a single CDR3 amino acid substitution. Finally, I studied how the dynamic flexibility of pMHC could be influenced by the peptide cargo by using red-edge excitation shift to examine a collection of well-studied analogues of a preproinsulin-derived peptide in the context of HLA A*0201. Conclusions – My research highlights both the negative (viral/autoimmunity) and the positive (viral/cancer) aspects of T cell crossreactivity and demonstrates that plasticity in both the TCR and cognate pMHC ligands are likely to play a role in the range of TCR-pMHC interactions that are capable of triggering a T cell response

    Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

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    The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers.DFG, 389687267, Kompartimentalisierung, Aufrechterhaltung und Reaktivierung humaner Gedächtnis-T-Lymphozyten aus Knochenmark und peripherem BlutDFG, 80750187, SFB 841: Leberentzündungen: Infektion, Immunregulation und KonsequenzenEC/H2020/800924/EU/International Cancer Research Fellowships - 2/iCARE-2DFG, 252623821, Die Rolle von follikulären T-Helferzellen in T-Helferzell-Differenzierung, Funktion und PlastizitätDFG, 390873048, EXC 2151: ImmunoSensation2 - the immune sensory syste

    Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

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    The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers

    Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

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    These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion

    Advances in Molecular Simulation

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    Molecular simulations are commonly used in physics, chemistry, biology, material science, engineering, and even medicine. This book provides a wide range of molecular simulation methods and their applications in various fields. It reflects the power of molecular simulation as an effective research tool. We hope that the presented results can provide an impetus for further fruitful studies
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