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3. Ammonia inhibits antitumor activity of NK cells by decreasing mature perforin

Author

Winiarska, Magdalena, primary, Domagala, Joanna, additional, Grzywa, Tomasz M., additional, Baranowska, Iwona, additional, Kusowska, Aleksandra, additional, Fidyt, Klaudyna, additional, Marhelava, Katsiaryna, additional, Pilch, Zofia, additional, Graczyk-Jarzynka, Agnieszka, additional, Picard, Lea, additional, Jastrzebski, Kamil, additional, Granica, Monika, additional, Justyniarska, Magdalena, additional, Urlaub, Doris, additional, Bobrowicz, Malgorzata, additional, Miaczynska, Marta, additional, and Watzl, Carsten, additional

Published
2023
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6. NK Cell Activation

Author

Claus, Maren, Urlaub, Doris, Watzl, Carsten, Mackay, Ian R., editor, Rose, Noel R., editor, Diamond, Betty, editor, and Davidson, Anne, editor

Published
2014
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7. In-depth analysis of T cell immunity and antibody responses in heterologous prime-boost-boost vaccine regimens against SARS-CoV-2 and Omicron variant

Author

Heinen, Natalie, primary, Marheinecke, Corinna Sophie, additional, Bessen, Clara, additional, Blazquez-Navarro, Arturo, additional, Roch, Toralf, additional, Stervbo, Ulrik, additional, Anft, Moritz, additional, Plaza-Sirvent, Carlos, additional, Busse, Sandra, additional, Klöhn, Mara, additional, Schrader, Jil, additional, Vidal Blanco, Elena, additional, Urlaub, Doris, additional, Watzl, Carsten, additional, Hoffmann, Markus, additional, Pöhlmann, Stefan, additional, Tenbusch, Matthias, additional, Steinmann, Eike, additional, Todt, Daniel, additional, Hagenbeck, Carsten, additional, Zimmer, Gert, additional, Schmidt, Wolfgang Ekkehard, additional, Quast, Daniel Robert, additional, Babel, Nina, additional, Schmitz, Ingo, additional, and Pfänder, Stephanie, additional

Published
2022
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8. Impact of SARS-CoV-2 vaccination on systemic immune responses in people living with HIV

Author

Bessen, Clara, primary, Plaza-Sirvent, Carlos, additional, Simsek, Agit, additional, Bhat, Jaydeep, additional, Marheinecke, Corinna, additional, Urlaub, Doris, additional, Bonowitz, Petra, additional, Busse, Sandra, additional, Schumann, Sabrina, additional, Blanco, Elena Vidal, additional, Skaletz-Rorowski, Adriane, additional, Brockmeyer, Norbert H., additional, Overheu, Oliver, additional, Reinacher-Schick, Anke, additional, Faissner, Simon, additional, Watzl, Carsten, additional, Pfaender, Stephanie, additional, Potthoff, Anja, additional, and Schmitz, Ingo, additional

Published
2022
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9. Immune response in ofatumumab treated multiple sclerosis patients after SARS-CoV-2 vaccination

Author

Faissner, Simon, primary, Heitmann, Neele, additional, Plaza-Sirvent, Carlos, additional, Trendelenburg, Paulina, additional, Ceylan, Ulas, additional, Motte, Jeremias, additional, Bessen, Clara, additional, Urlaub, Doris, additional, Watzl, Carsten, additional, Overheu, Oliver, additional, Reinacher-Schick, Anke, additional, Hellwig, Kerstin, additional, Pfaender, Stephanie, additional, Schmitz, Ingo, additional, and Gold, Ralf, additional

Published
2022
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11. Neutralizing antibody responses 300 days after SARS‐CoV‐2 infection and induction of high antibody titers after vaccination

Author

Urlaub, Doris, primary, Wolfsdorff, Natalie, additional, Hoffmann, Jan‐Erik, additional, Dorok, Stefanie, additional, Hoffmann, Markus, additional, Anft, Moritz, additional, Pieris, Naomi, additional, Günther, Patrick, additional, Schaaf, Bernhard, additional, Cassens, Uwe, additional, Bröde, Peter, additional, Claus, Maren, additional, Picard, Lea K., additional, Wingert, Sabine, additional, Backes, Simone, additional, Durak, Deniz, additional, Babel, Nina, additional, Pöhlmann, Stefan, additional, Renken, Frank, additional, Raunser, Stefan, additional, and Watzl, Carsten, additional

Published
2022
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13. Immune response in ofatumumab treated multiple sclerosis patients aft er SARS-CoV-2 vaccination.

Author

Faissner, Simon, Heitmann, Neele, Plaza-Sirvent, Carlos, Trendelenburg, Paulina, Ceylan, Ulas, Motte, Jeremias, Bessen, Clara, Urlaub, Doris, Watzl, Carsten, Overheu, Oliver, Reinacher-Schick, Anke, Hellwig, Kerstin, Pfaender, Stephanie, Schmitz, Ingo, and Gold, Ralf

Subjects
IMMUNE response, HUMORAL immunity, REGULATORY T cells, MULTIPLE sclerosis, SARS-CoV-2
Abstract

Objective: The pandemic induced by SARS-CoV-2 has huge implications for patients with immunosuppression that is caused by disorders or specific treatments. Especially approaches targeting B cells via anti-CD20 therapy are associated with impaired humoral immune response but sustained cellular immunity. Ofatumumab is a human anti-CD20 directed antibody applied in low dosages subcutaneously, recently licensed for Multiple Sclerosis (MS). Effects of early ofatumumab treatment on alterations of immune cell composition and immune response towards SARS-CoV-2 are incompletely understood. Methods: We here investigated immune cell alterations in early ofatumumab (Ofa) treated patients and effects on humoral (titer, neutralization capacity against wild type, Delta and Omicron) and cellular immune responses in Ofa treated MS patients following a third vaccination against SARS-CoV-2 compared to healthy controls. Results: We show that a mean treatment duration of three months in the Ofa group led to near complete B cell depletion in line with altered composition of certain CD4+ T cell subpopulations such as enhanced frequencies of naive and a decrease of non-suppressive regulatory T cells (Tregs). Titer and neutralization capacity against SARS-CoV-2 variants was impaired while cellular immune response was sustained, characterized by a strong T helper 1 profile (Th1). Interpretation: In summary, low dosage ofatumumab treatment elicits sustained depletion of B cells in line with alterations of immune cells, mainly Tregs. This is associated with impaired humoral immune response towards SARS-CoV-2 vaccination but preserved, Th1 driven cellular immunity adding crucial information regarding early effects of low dosage anti-CD20 therapy on humoral and cellular immunity. [ABSTRACT FROM AUTHOR]

Published
2022
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14. Humoral and cellular immune responses to repeated SARS‐CoV‐2 infection and vaccination in an individual living with HIV.

Author

Simsek, Agit, Bessen, Clara, Meister, Toni L, Urlaub, Doris, Skaletz‐Rorowski, Adriane, Brockmeyer, Norbert H, Overheu, Oliver, Reinacher‐Schick, Anke, Faissner, Simon, Watzl, Carsten, Pfaender, Stephanie, Schmitz, Ingo, Potthoff, Anja, and Plaza‐Sirvent, Carlos

Subjects
SARS-CoV-2, VACCINATION, REGULATORY T cells, SARS-CoV-2 Delta variant, HIV
Abstract

Similarly, while patient's plasma was not able to neutralize SARS-CoV-2 in vitro following first infection, the WT variant was strongly neutralized after the first vaccination. Humoral and cellular immune responses to repeated SARS-CoV-2 infection and vaccination in an individual living with HIV T cells targeting SARS-CoV-2: by infection, vaccination, and against future variants. [Extracted from the article]

Published
2023
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19. Single-Fluorescent Protein Reporters Allow Parallel Quantification of Natural Killer Cell-Mediated Granzyme and Caspase Activities in Single Target Cells

Author

Liesche, Clarissa, Sauer, Patricia, Prager, Isabel, Urlaub, Doris, Claus, Maren, Eils, Roland, Beaudouin, Joel, and Watzl, Carsten

Subjects
lcsh:Immunologic diseases. Allergy, apoptosis and cell death, natural killer cells, Cell Death, Immunology, Gene Expression, Apoptosis, Granzymes, single-fluorescent protein reporters, granzyme and caspase activity, Killer Cells, Natural, cytotoxic lymphocytes, Genes, Reporter, Caspases, Proteolysis, Methods, Humans, Single-Cell Analysis, lcsh:RC581-607
Abstract

Natural killer (NK) cells eliminate infected and tumorigenic cells through delivery of granzymes via perforin pores or by activation of caspases via death receptors. In order to understand how NK cells combine different cell death mechanisms, it is important to quantify target cell responses on a single cell level. However, currently existing reporters do not allow the measurement of several protease activities inside the same cell. Here, we present a strategy for the comparison of two different proteases at a time inside individual target cells upon engagement by NK cells. We developed single-fluorescent protein reporters containing the RIEAD or the VGPD cleavage site for the measurement of granzyme B activity. We show that these two granzyme B reporters can be applied in combination with caspase-8 or caspase-3 reporters. While we did not find that caspase-8 was activated by granzyme B, our method revealed that caspase-3 activity follows granzyme B activity with a delay of about 6 min. Finally, we illustrate the comparison of several different reporters for granzyme A, M, K, and H. The approach presented here is a valuable means for the investigation of the temporal evolution of cell death mediated by cytotoxic lymphocytes.

Published
2018

22. NK cells switch from granzyme B to death receptor–mediated cytotoxicity during serial killing

Author

Prager, Isabel, Liesche, Clarissa, van Ooijen, Hanna, Urlaub, Doris, Verron, Quentin, Sandström, Niklas, Fasbender, Frank, Claus, Maren, Eils, Roland, Beaudouin, Joël, Önfelt, Björn, Watzl, Carsten, Prager, Isabel, Liesche, Clarissa, van Ooijen, Hanna, Urlaub, Doris, Verron, Quentin, Sandström, Niklas, Fasbender, Frank, Claus, Maren, Eils, Roland, Beaudouin, Joël, Önfelt, Björn, and Watzl, Carsten

Abstract

NK cells eliminate virus-infected and tumor cells by releasing cytotoxic granules containing granzyme B (GrzB) or by engaging death receptors that initiate caspase cascades. The orchestrated interplay between both cell death pathways remains poorly defined. Here we simultaneously measure the activities of GrzB and caspase-8 in tumor cells upon contact with human NK cells. We observed that NK cells switch from inducing a fast GrzB-mediated cell death in their first killing events to a slow death receptor–mediated killing during subsequent tumor cell encounters. Target cell contact reduced intracellular GrzB and perforin and increased surface-CD95L in NK cells over time, showing how the switch in cytotoxicity pathways is controlled. Without perforin, NK cells were unable to perform GrzB-mediated serial killing and only killed once via death receptors. In contrast, the absence of CD95 on tumor targets did not impair GrzB-mediated serial killing. This demonstrates that GrzB and death receptor–mediated cytotoxicity are differentially regulated during NK cell serial killing., QC 20190822

Published
2019
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26. Molecular analysis of the decision making process in NK cells

Author

Urlaub, Doris

Subjects
570 Life sciences
Abstract

Natural killer (NK) cells are a subpopulation of lymphocytes that are involved in the control of different tumors and infections. Unlike T and B cells, NK cells belong to the innate part of the immune system. NK cells carry on their surface a multitude of activating and inhibitory receptors. The regulation of NK cell activation depends on a balance of positive and negative signals initiated by various receptors. Triggering of activating receptors leads to Src family kinase mediated Vav1 phosphorylation, whereas inhibitory receptors dephosphorylate Vav1 via the phosphatase SHP-1. This makes Vav1 the first point where negative signals can intercept the activating signaling cascade. In cooperation with computational biologists we established a mathematical model describing these early signaling events to gain insight into the integration of positive and negative signals on a molecular level. In quantitative mathematical models each equation refers to identifiable processes and parameters have physical interpretation (such as concentration, binding affinity and reaction rate). Therefore we quantified the concentrations of involved molecules. The predictions from the model and our experimental data show that engagement of activating receptors results in a rapid switch-like increase of Vav1 phosphorylation. Similarly, engagement of inhibitory receptors induces a switch-like dephosphorylation of Vav1 that is dominant over activating signals. Comparing experimental results to predictions derived from a family of simplified models shows that kinase association with the NK cell receptors and the enhanced activity of SHP-1 bound to inhibitory receptors is essential to simulate such a physiological response. Interestingly, other concepts of immune receptor signaling such as phosphatase segregation and kinase autophosphorylation were dispensable for our model. The cytotoxic activity of NK cells induced by a combination of activating and inhibitory signals correlates with the switch-like Vav1 phosphorylation. Our data are consistent with a central role of Vav1 in the decision making process of NK cells and enable a novel insight into the integration of positive and negative signals during lymphocyte activation.

Published
2009

31. Molecular mechanisms of natural killer cell regulation.

Author

Watzl C and Urlaub D

Subjects
Humans, Lymphocyte Activation, Killer Cells, Natural immunology
Abstract

Natural Killer (NK) cells are important for early immune reactions against viral infections and cancer. They are regulated by a highly redundant system of different activating and inhibitory receptors. Here we summarize our current understanding about the regulation of these cells and describe how mathematical modeling and systems biology approaches can help to shed some light on the complex regulatory network that governs NK cell reactivity.

Published
2012
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