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Article ; Online: Identification of molecular candidates which regulate calcium-dependent CD8

Zöphel, Sylvia / Schäfer, Gertrud / Nazarieh, Maryam / Konetzki, Verena / Hoxha, Cora / Meese, Eckart / Hoth, Markus / Helms, Volkhard / Hamed, Mohamed / Schwarz, Eva C

Molecular immunology

2023 Volume 157, Page(s) 202–213

Abstract: Cytotoxic ... ...

Abstract	Cytotoxic CD8
MeSH term(s)	Humans ; CD8-Positive T-Lymphocytes ; Calcium ; Cytotoxicity, Immunologic ; T-Lymphocytes, Cytotoxic ; Killer Cells, Natural ; Antineoplastic Agents
Chemical Substances	Calcium (SY7Q814VUP) ; Antineoplastic Agents
Language	English
Publishing date	2023-04-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2023.04.002
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Article ; Online: Cytotoxic Efficiency of Human CD8

Knörck, Arne / Schäfer, Gertrud / Alansary, Dalia / Richter, Josephine / Thurner, Lorenz / Hoth, Markus / Schwarz, Eva C

Frontiers in immunology

2022 Volume 13, Page(s) 838484

Abstract: Immunological memory is important to protect humans against recurring diseases. Memory ... ...

Abstract	Immunological memory is important to protect humans against recurring diseases. Memory CD8
MeSH term(s)	Antineoplastic Agents ; CD8-Positive T-Lymphocytes ; Humans ; Immunologic Memory ; Necrosis/metabolism ; Neoplasm Recurrence, Local/metabolism ; Perforin/metabolism
Chemical Substances	Antineoplastic Agents ; Perforin (126465-35-8)
Language	English
Publishing date	2022-04-13
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2022.838484
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Article ; Conference proceedings: Die Quantifizierung von Koronarkalk auf Grundlage virtuell nativer Rekonstruktionen aus koronaren CT-Angiographien an einem Photon-Counting Detektor CT

Risch, F / Braun, F / Decker, J / Eva, H / Scheurig-Münkler, C / Schwarz, E / Schwarz, F / Kröncke, T / Schoepf, J / Woznicki, P / Wolny, C

RöFo - Fortschritte auf dem Gebiet der Röntgenstrahlen und der bildgebenden Verfahren

2022 Volume 194, Issue S 01

Event/congress	103. Deutscher Röntgenkongress der Deutschen Röntgengesellschaft e. V., RheinMain CongressCenter Wiesbaden, 2022-05-25
Language	German
Publishing date	2022-08-01
Publisher	Georg Thieme Verlag
Publishing place	Stuttgart ; New York
Document type	Article ; Conference proceedings
ZDB-ID	554830-5
ISSN	1438-9010 ; 1438-9029 ; 0340-1618 ; 0936-6652 ; 1433-5972
ISSN (online)	1438-9010
ISSN	1438-9029 ; 0340-1618 ; 0936-6652 ; 1433-5972
DOI	10.1055/s-0042-1749770
Database	Thieme publisher's database

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Article ; Online: Identification of molecular candidates which regulate calcium-dependent CD8+ T-cell cytotoxicity

Zöphel, Sylvia / Schäfer, Gertrud / Nazarieh, Maryam / Konetzki, Verena / Hoxha, Cora / Meese, Eckart / Hoth, Markus / Helms, Volkhard / Abū al-Saʻūd, Muḥammad Ḥāmid Muʻawwaḍ / Schwarz, Eva C.

Molecular Immunology. 2023 May, v. 157 p.202-213

2023

Abstract: ... Ca²⁺-limiting conditions. Overall, we identified four hits, CCR5 (C-C chemokine receptor type five ...

Abstract	Cytotoxic CD8⁺ T lymphocytes (CTL) eliminate infected cells or transformed tumor cells by releasing perforin-containing cytotoxic granules at the immunological synapse. The secretion of such granules depends on Ca²⁺-influx through store operated Ca²⁺ channels, formed by STIM (stromal interaction molecule)-activated Orai proteins. Whereas molecular mechanisms of the secretion machinery are well understood, much less is known about the molecular machinery that regulates the efficiency of Ca²⁺-dependent target cell killing. CTL killing efficiency is of high interest considering the number of studies on CD8⁺ T lymphocytes modified for clinical use. Here, we isolated total RNA from primary human cells: natural killer (NK) cells, non-stimulated CD8⁺ T-cells, and from Staphylococcus aureus enterotoxin A (SEA) stimulated CD8⁺ T-cells (SEA-CTL) and conducted whole genome expression profiling by microarray experiments. Based on differential expression analysis of the transcriptome data and analysis of master regulator genes, we identified 31 candidates which potentially regulate Ca²⁺-homeostasis in CTL. To investigate a putative function of these candidates in CTL cytotoxicity, we transfected either SEA-stimulated CTL (SEA-CTL) or antigen specific CD8⁺ T-cell clones (CTL-MART-1) with siRNAs specific against the identified candidates and analyzed the killing capacity using a real-time killing assay. In addition, we complemented the analysis by studying the effect of inhibitory substances acting on the candidate proteins if available. Finally, to unmask their involvement in Ca²⁺ dependent cytotoxicity, candidates were also analyzed under Ca²⁺-limiting conditions. Overall, we identified four hits, CCR5 (C-C chemokine receptor type five), KCNN4 (potassium calcium-activated channel subfamily N), RCAN3 (regulator of calcineurin) and BCL (B-cell lymphoma) 2 which clearly affect the efficiency of Ca²⁺ dependent cytotoxicity in CTL-MART-1 cells, CCR5, BCL2, and KCNN4 in a positive manner, and RCAN3 in a negative way.
Keywords	B-cell lymphoma ; CD8-positive T-lymphocytes ; RNA ; Staphylococcus aureus ; antigens ; beta chemokines ; calcium ; chemokine receptors ; cytotoxicity ; enterotoxins ; gene expression regulation ; genome ; humans ; immunological synapse ; microarray technology ; potassium ; secretion ; transcriptome ; CTL (cytotoxic T lymphocytes) ; Real-time killing assay ; Cytotoxic efficiency ; Differential expression analyses ; Transcriptome data analyses
Language	English
Dates of publication	2023-05
Size	p. 202-213.
Publishing place	Elsevier Ltd
Document type	Article ; Online
Note	Use and reproduction
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2023.04.002
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: High glucose distinctively regulates Ca

Zou, Huajiao / Yang, Wenjuan / Schwär, Gertrud / Zhao, Renping / Alansary, Dalia / Yin, Deling / Schwarz, Eva C / Niemeyer, Barbara A / Qu, Bin

European journal of immunology

2020 Volume 50, Issue 12, Page(s) 2095–2098

Abstract: In CTLs: High glucose-culture enhances thapsigargin-induced SOCE but decreases target recognition-induced ... ...

Abstract	In CTLs: High glucose-culture enhances thapsigargin-induced SOCE but decreases target recognition-induced Ca
MeSH term(s)	Calcium/metabolism ; Calcium Channels/metabolism ; Glucose/metabolism ; Humans ; Signal Transduction/drug effects ; T-Lymphocytes, Cytotoxic/drug effects ; T-Lymphocytes, Cytotoxic/metabolism ; Thapsigargin/pharmacology
Chemical Substances	Calcium Channels ; Thapsigargin (67526-95-8) ; Glucose (IY9XDZ35W2) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2020-08-18
Publishing country	Germany
Document type	Letter ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202048577
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Article ; Online: Interdependence of sequential cytotoxic T lymphocyte and natural killer cell cytotoxicity against melanoma cells.

Friedmann, Kim S / Kaschek, Lea / Knörck, Arne / Cappello, Sabrina / Lünsmann, Niklas / Küchler, Nadja / Hoxha, Cora / Schäfer, Gertrud / Iden, Sandra / Bogeski, Ivan / Kummerow, Carsten / Schwarz, Eva C / Hoth, Markus

The Journal of physiology

2022 Volume 600, Issue 23, Page(s) 5027–5054

Abstract: Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells recognize and eliminate cancer cells. However, immune evasion, downregulation of immune function by the tumour microenvironment and resistance of cancer cells are major problems. Although CTL ... ...

Abstract	Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells recognize and eliminate cancer cells. However, immune evasion, downregulation of immune function by the tumour microenvironment and resistance of cancer cells are major problems. Although CTL and NK cells are both important to eliminate cancer, most studies address them individually. We quantified sequential primary human CTL and NK cell cytotoxicity against the melanoma cell line SK-Mel-5. At high effector-to-target ratios, NK cells or melan-A (MART-1)-specific CTL eliminated all SK-Mel-5 cells within 24 h, indicating that SK-Mel-5 cells are not resistant initially. However, at lower effector-to-target ratios, which resemble numbers of the immune contexture in human cancer, a substantial number of SK-Mel-5 cells survived. Pre-exposure to CTL induced resistance in surviving SK-Mel-5 cells to subsequent CTL or NK cell cytotoxicity, and pre-exposure to NK cells induced resistance in surviving SK-Mel-5 cells to NK cells. Higher human leucocyte antigen class I expression or interleukin-6 levels were correlated with resistance to NK cells, whereas reduction in MART-1 antigen expression was correlated with reduced CTL cytotoxicity. The CTL cytotoxicity was rescued beyond control levels by exogenous MART-1 antigen. In contrast to the other three combinations, CTL cytotoxicity against SK-Mel-5 cells was enhanced following NK cell pre-exposure. Our assay allows quantification of sequential CTL and NK cell cytotoxicity and might guide strategies for efficient CTL-NK cell anti-melanoma therapies. KEY POINTS: Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells eliminate cancer cells. Both CTL and NK cells attack the same targets, but most studies address them individually. In a sequential cytotoxicity model, the interdependence of antigen-specific CTL and NK cell cytotoxicity against melanoma is quantified. High numbers of antigen-specific CTL and NK cells eliminate all melanoma cells. However, lower numbers induce resistance if secondary CTL or NK cell exposure follows initial CTL exposure or if secondary NK cell exposure follows initial NK cell exposure. On the contrary, if secondary CTL exposure follows initial NK cell exposure, cytotoxicity is enhanced. Alterations in human leucocyte antigen class I expression and interleukin-6 levels are correlated with resistance to NK cells, whereas a reduction in antigen expression is correlated with reduced CTL cytotoxicity; CTL cytotoxicity is rescued beyond control levels by exogenous antigen. This assay and the results on interdependencies will help us to understand and optimize immune therapies against cancer.
MeSH term(s)	Humans ; T-Lymphocytes, Cytotoxic ; MART-1 Antigen ; Interleukin-6 ; Killer Cells, Natural ; Melanoma ; Tumor Microenvironment
Chemical Substances	MART-1 Antigen ; Interleukin-6
Language	English
Publishing date	2022-11-03
Publishing country	England
Document type	Journal Article
ZDB-ID	3115-x
ISSN	1469-7793 ; 0022-3751
ISSN (online)	1469-7793
ISSN	0022-3751
DOI	10.1113/JP283667
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Article ; Online: Compound heterozygous variants in OTULIN are associated with fulminant atypical late-onset ORAS.

EMBO molecular medicine

2022 Volume 14, Issue 3, Page(s) e14901

Abstract: ... heterozygous variants (c.258G>A (p.M86I)/c.500G>C (p.W167S)) in the OTULIN gene in a 7-year-old affected ...

Abstract	Autoinflammatory diseases are a heterogenous group of disorders defined by fever and systemic inflammation suggesting involvement of genes regulating innate immune responses. Patients with homozygous loss-of-function variants in the OTU-deubiquitinase OTULIN suffer from neonatal-onset OTULIN-related autoinflammatory syndrome (ORAS) characterized by fever, panniculitis, diarrhea, and arthritis. Here, we describe an atypical form of ORAS with distinct clinical manifestation of the disease caused by two new compound heterozygous variants (c.258G>A (p.M86I)/c.500G>C (p.W167S)) in the OTULIN gene in a 7-year-old affected by a life-threatening autoinflammatory episode with sterile abscess formation. On the molecular level, we find binding of OTULIN to linear ubiquitin to be compromised by both variants; however, protein stability and catalytic activity is most affected by OTULIN variant p.W167S. These molecular changes together lead to increased levels of linear ubiquitin linkages in patient-derived cells triggering the disease. Our data indicate that the spectrum of ORAS patients is more diverse than previously thought and, thus, supposedly asymptomatic individuals might also be affected. Based on our results, we propose to subdivide the ORAS into classical and atypical entities.
MeSH term(s)	Child ; Endopeptidases/genetics ; Hereditary Autoinflammatory Diseases/genetics ; Humans ; Infant, Newborn ; Inflammation/genetics ; Ubiquitin/metabolism
Chemical Substances	Ubiquitin ; Endopeptidases (EC 3.4.-) ; OTULIN protein, human (EC 3.4.-)
Language	English
Publishing date	2022-02-16
Publishing country	England
Document type	Case Reports ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2467145-9
ISSN	1757-4684 ; 1757-4676
ISSN (online)	1757-4684
ISSN	1757-4676
DOI	10.15252/emmm.202114901
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Article ; Online: High Glucose Enhances Cytotoxic T Lymphocyte-Mediated Cytotoxicity.

Zhu, Jie / Yang, Wenjuan / Zhou, Xiangda / Zöphel, Dorina / Soriano-Baguet, Leticia / Dolgener, Denise / Carlein, Christopher / Hof, Chantal / Zhao, Renping / Ye, Shandong / Schwarz, Eva C / Brenner, Dirk / Prates Roma, Leticia / Qu, Bin

Frontiers in immunology

2021 Volume 12, Page(s) 689337

Abstract: Cytotoxic T lymphocytes (CTLs) are key players to eliminate tumorigenic or pathogen-infected cells using lytic granules (LG) and Fas ligand (FasL) pathways. Depletion of glucose leads to severely impaired cytotoxic function of CTLs. However, the impact ... ...

Abstract	Cytotoxic T lymphocytes (CTLs) are key players to eliminate tumorigenic or pathogen-infected cells using lytic granules (LG) and Fas ligand (FasL) pathways. Depletion of glucose leads to severely impaired cytotoxic function of CTLs. However, the impact of excessive glucose on CTL functions still remains largely unknown. Here we used primary human CD8
MeSH term(s)	Animals ; Cells, Cultured ; Glucose/pharmacology ; Glycolysis/drug effects ; Humans ; Male ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic/drug effects ; T-Lymphocytes, Cytotoxic/metabolism ; Mice
Chemical Substances	Glucose (IY9XDZ35W2)
Language	English
Publishing date	2021-06-25
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2021.689337
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Article ; Online: Macrophage Polarization is Deregulated in Haemophilia.

Knowles, Lynn M / Kagiri, Daniela / Bernard, Martin / Schwarz, Eva C / Eichler, Hermann / Pilch, Jan

Thrombosis and haemostasis

2019 Volume 119, Issue 2, Page(s) 234–245

Abstract: ... of haemophilia patients, to a monocyte cell line. The increase of leptin occurred in conjunction with C-reactive ...

Abstract	Macrophages make important contributions to inflammation and wound healing. We show here that macrophage polarization is deregulated in haemophilia in response to macrophage colony-stimulating factor (M-CSF) and partially in response to granulocyte-macrophage colony-stimulating factor (GM-CSF). As a result, haemophilia macrophages exhibit a specific impairment of M-CSF-mediated functions involved in wound healing such as clot invasion and phagocytosis. Haemophilia monocytes express reduced amounts of the receptors for M-CSF and GM-CSF, which correlates with a failure to express tumour necrosis factor α (TNFα) and CD163 in M-CSF-treated haemophilia macrophages and reduced expression of TNFα and CD206 after treatment with GM-CSF. Protein expression in response to M-CSF was regained with respect to CD163 and CD206 after embedding haemophilia monocytes in clotted plasma suggesting that a functioning coagulation system has positive effects on macrophage M2 polarization. Mimicking the functional deficits of haemophilia macrophages in normal macrophages was possible by adding leptin, which we found to be elevated in the blood of haemophilia patients, to a monocyte cell line. The increase of leptin occurred in conjunction with C-reactive protein in a body mass index-controlled cohort suggesting that haemophilia patients harbour chronic low-grade inflammation. Together, our data indicate that impaired clotting in haemophilia patients leads to increased inflammation and a deregulation in macrophage differentiation, which may explain the commonly observed deficits in wound healing and tissue regeneration.
MeSH term(s)	Adult ; Antigens, CD/metabolism ; Antigens, Differentiation, Myelomonocytic/metabolism ; Blood Circulation ; Blood Coagulation ; Cell Differentiation ; Cohort Studies ; Enzyme-Linked Immunosorbent Assay ; Erythrocytes/cytology ; Fibrin/metabolism ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor/therapeutic use ; Hemophilia A/blood ; Hemophilia A/drug therapy ; Humans ; Lectins, C-Type/metabolism ; Leukocytes, Mononuclear/cytology ; Macrophage Colony-Stimulating Factor/therapeutic use ; Macrophages/cytology ; Male ; Mannose-Binding Lectins/metabolism ; Microscopy, Fluorescence ; Monocytes/cytology ; Phagocytosis ; Receptors, Cell Surface/metabolism ; Tumor Necrosis Factor-alpha/metabolism ; Young Adult
Chemical Substances	Antigens, CD ; Antigens, Differentiation, Myelomonocytic ; CD163 antigen ; Lectins, C-Type ; Mannose-Binding Lectins ; Receptors, Cell Surface ; Tumor Necrosis Factor-alpha ; mannose receptor ; Macrophage Colony-Stimulating Factor (81627-83-0) ; Granulocyte-Macrophage Colony-Stimulating Factor (83869-56-1) ; Fibrin (9001-31-4)
Language	English
Publishing date	2019-01-16
Publishing country	Germany
Document type	Journal Article
ZDB-ID	518294-3
ISSN	2567-689X ; 0340-6245
ISSN (online)	2567-689X
ISSN	0340-6245
DOI	10.1055/s-0038-1676796
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Article ; Online: Unspecific CTL Killing Is Enhanced by High Glucose

Yang, Wenjuan / Denger, Andreas / Diener, Caroline / Küppers, Frederic / Soriano-Baguet, Leticia / Schäfer, Gertrud / Yanamandra, Archana K / Zhao, Renping / Knörck, Arne / Schwarz, Eva C / Hart, Martin / Lammert, Frank / Roma, Leticia Prates / Brenner, Dirk / Christidis, Grigorios / Helms, Volkhard / Meese, Eckart / Hoth, Markus / Qu, Bin

Frontiers in immunology

2022 Volume 13, Page(s) 831680

Abstract: TNF-related apoptosis inducing ligand (TRAIL) is expressed on cytotoxic T lymphocytes (CTLs) and TRAIL is linked to progression of diabetes. However, the impact of high glucose on TRAIL expression and its related killing function in CTLs still remains ... ...

Abstract	TNF-related apoptosis inducing ligand (TRAIL) is expressed on cytotoxic T lymphocytes (CTLs) and TRAIL is linked to progression of diabetes. However, the impact of high glucose on TRAIL expression and its related killing function in CTLs still remains largely elusive. Here, we report that TRAIL is substantially up-regulated in CTLs in environments with high glucose (HG) both
MeSH term(s)	Glucose/metabolism ; Phosphatidylinositol 3-Kinases/metabolism ; Receptors, TNF-Related Apoptosis-Inducing Ligand/metabolism ; T-Lymphocytes, Cytotoxic/metabolism ; TNF-Related Apoptosis-Inducing Ligand/metabolism
Chemical Substances	Receptors, TNF-Related Apoptosis-Inducing Ligand ; TNF-Related Apoptosis-Inducing Ligand ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2022-02-21
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2022.831680
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