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  1. Article ; Online: T cell-intrinsic protein kinase D3 is dispensable for the cells' activation.

    Koutník, Jiří / Leitges, Michael / Siegmund, Kerstin

    Frontiers in immunology

    2022  Volume 13, Page(s) 1049033

    Abstract: Protein kinases D (PKDs) are implicated in T cell receptor (TCR) signaling. Of the two T cell-expressed isoforms PKD2 and PKD3, however, only the former one is rather well understood in this immune cell type. Recently, we have observed a putative hyper- ... ...

    Abstract Protein kinases D (PKDs) are implicated in T cell receptor (TCR) signaling. Of the two T cell-expressed isoforms PKD2 and PKD3, however, only the former one is rather well understood in this immune cell type. Recently, we have observed a putative hyper-phenotype of T cells from conventional PKD3-knockout mice, which we explained as a secondary effect due to a skewed T cell compartment from naïve towards effector/memory T cells already under steady state conditions. Nonetheless, to this end it is not clear whether these aberrations are mediated by a T cell-intrinsic or -extrinsic function of PKD3. To address this question, we have investigated mice lacking PKD3 specifically in the T cell compartment. We could show that T cells from CD4-Cre-driven conditional knockout mice did not phenocopy the ones from conventional PKD3-knockout mice. In brief, no skewing in the T cell compartment of peripheral lymphoid organs, no hyper-activation upon stimulation
    MeSH term(s) Mice ; Animals ; Lymphocyte Count ; Germinal Center ; Mice, Knockout ; Receptors, Antigen, T-Cell/genetics
    Chemical Substances protein kinase C nu (EC 2.7.1.-) ; Receptors, Antigen, T-Cell
    Language English
    Publishing date 2022-11-17
    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.2022.1049033
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Emerging Next-Generation Target for Cancer Immunotherapy Research: The Orphan Nuclear Receptor NR2F6.

    Klepsch, Victoria / Siegmund, Kerstin / Baier, Gottfried

    Cancers

    2021  Volume 13, Issue 11

    Abstract: Additional therapeutic targets suitable for boosting anti-tumor effector responses have been found inside effector ... ...

    Abstract Additional therapeutic targets suitable for boosting anti-tumor effector responses have been found inside effector CD4
    Language English
    Publishing date 2021-05-26
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2527080-1
    ISSN 2072-6694
    ISSN 2072-6694
    DOI 10.3390/cancers13112600
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Chemically modified mRNA nucleofection of primary human T cells.

    Thuille, Nikolaus / Sajinovic, Tajana / Siegmund, Kerstin / Baier, Gottfried

    Journal of immunological methods

    2020  Volume 487, Page(s) 112878

    Abstract: Here we show that an approach of in-vitro transcribed mRNA nucleofection expands the range of transfection of primary human T cells. It represents a reproducible and time-efficient technology, and is thus an ideal tool in basic research involving highly ... ...

    Abstract Here we show that an approach of in-vitro transcribed mRNA nucleofection expands the range of transfection of primary human T cells. It represents a reproducible and time-efficient technology, and is thus an ideal tool in basic research involving highly controlled in-vitro experiments with a gene of interest aiming at identifying its biological human T cell function.
    MeSH term(s) Green Fluorescent Proteins/biosynthesis ; Green Fluorescent Proteins/genetics ; Humans ; Jurkat Cells ; RNA, Messenger/biosynthesis ; RNA, Messenger/chemistry ; RNA, Messenger/genetics ; Repressor Proteins/biosynthesis ; Repressor Proteins/genetics ; T-Lymphocytes/metabolism ; Time Factors ; Transcription, Genetic ; Transfection
    Chemical Substances NR2F6 protein, human ; RNA, Messenger ; Repressor Proteins ; enhanced green fluorescent protein ; Green Fluorescent Proteins (147336-22-9)
    Language English
    Publishing date 2020-10-06
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2020.112878
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 795: Show issues Location:
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    Jg. 1995 - 2021: Lesesall (1.OG)
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  4. Article: Chemically modified mRNA nucleofection of primary human T cells

    Thuille, Nikolaus / Sajinovic, Tajana / Siegmund, Kerstin / Baier, Gottfried

    Journal of immunological methods. 2020 Dec., v. 487

    2020  

    Abstract: Here we show that an approach of in-vitro transcribed mRNA nucleofection expands the range of transfection of primary human T cells. It represents a reproducible and time-efficient technology, and is thus an ideal tool in basic research involving highly ... ...

    Abstract Here we show that an approach of in-vitro transcribed mRNA nucleofection expands the range of transfection of primary human T cells. It represents a reproducible and time-efficient technology, and is thus an ideal tool in basic research involving highly controlled in-vitro experiments with a gene of interest aiming at identifying its biological human T cell function.
    Keywords T-lymphocytes ; genes ; humans ; in vitro studies ; messenger RNA ; transcription (genetics) ; transfection
    Language English
    Dates of publication 2020-12
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2020.112878
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 795: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
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    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  5. Article ; Online: A MLR-Based Approach to Analyze Regulators of T Lymphocyte Activation In Vivo.

    Koutník, Jiří / Klepsch, Victoria / Pommermayr, Maria / Thuille, Nikolaus / Baier, Gottfried / Siegmund, Kerstin

    International journal of molecular sciences

    2022  Volume 23, Issue 10

    Abstract: Depending on the context, robust and durable T lymphocyte activation is either desirable, as in the case of anti-tumor responses, or unwanted, in cases of autoimmunity when chronic stimulation leads to self-tissue damage. Therefore, reliable in vivo ... ...

    Abstract Depending on the context, robust and durable T lymphocyte activation is either desirable, as in the case of anti-tumor responses, or unwanted, in cases of autoimmunity when chronic stimulation leads to self-tissue damage. Therefore, reliable in vivo models are of great importance to identify and validate regulatory pathways of T lymphocyte activation. Here, we describe an in vivo mixed-lymphocyte-reaction (MLR) approach, which is based on the so-called parent-into-F1 (P → F1) mouse model in combination with the congenic marker CD45.1/2 and cell proliferation dye-labeling. This setup allows us to track adoptively transferred allogenic CD4
    MeSH term(s) Animals ; Lymphocyte Activation ; Lymphocyte Culture Test, Mixed ; Mice ; Spleen ; T-Lymphocytes
    Language English
    Publishing date 2022-05-10
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms23105337
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Loss of the orphan nuclear receptor NR2F6 enhances CD8

    Jakic, Bojana / Olson, William J / Siegmund, Kerstin / Klepsch, Victoria / Kimpel, Janine / Labi, Verena / Zehn, Dietmar / Baier, Gottfried / Hermann-Kleiter, Natascha

    Cell death & disease

    2021  Volume 12, Issue 2, Page(s) 187

    Abstract: Memory formation is a hallmark of T cell-mediated immunity, but how differentiation into either short-lived effector cells (SLECs, ... ...

    Abstract Memory formation is a hallmark of T cell-mediated immunity, but how differentiation into either short-lived effector cells (SLECs, CD127
    Language English
    Publishing date 2021-02-15
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2541626-1
    ISSN 2041-4889 ; 2041-4889
    ISSN (online) 2041-4889
    ISSN 2041-4889
    DOI 10.1038/s41419-021-03470-9
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Novel mutant mouse line emphasizes the importance of protein kinase C theta for CD4

    Siegmund, Kerstin / Thuille, Nikolaus / Posch, Nina / Fresser, Friedrich / Leitges, Michael / Baier, Gottfried

    Cell communication and signaling : CCS

    2019  Volume 17, Issue 1, Page(s) 56

    Abstract: Background: The protein kinase C theta (PKCθ) has an important and non-redundant function downstream of the antigen receptor and co-receptor complex in T lymphocytes. PKCθ is not only essential for activation of NF-κB, AP-1 and NFAT and subsequent ... ...

    Abstract Background: The protein kinase C theta (PKCθ) has an important and non-redundant function downstream of the antigen receptor and co-receptor complex in T lymphocytes. PKCθ is not only essential for activation of NF-κB, AP-1 and NFAT and subsequent interleukin-2 expression, but also critical for positive selection and development of regulatory T lymphocytes in the thymus. Several domains regulate its activity, such as a pseudosubstrate sequence mediating an auto-inhibitory intramolecular interaction, the tandem C1 domains binding diacylglycerol, and phosphorylation at conserved tyrosine, threonine as well as serine residues throughout the whole length of the protein. To address the importance of the variable domain V1 at the very N-terminus, which is encoded by exon 2, a mutated version of PKCθ was analyzed for its ability to stimulate T lymphocyte activation.
    Methods: T cell responses were analyzed with promoter luciferase reporter assays in Jurkat T cells transfected with PKCθ expression constructs. A mouse line expressing mutated instead of wild type PKCθ was analyzed in comparison to PKCθ-deficient and wild type mice for thymic development and T cell subsets by flow cytometry and T cell activation by quantitative RT-PCR, luminex analysis and flow cytometry.
    Results: In cell lines, the exon 2-replacing mutation impaired the transactivation of interleukin-2 expression by constitutively active mutant form of PKCθ. Moreover, analysis of a newly generated exon 2-mutant mouse line (PKCθ-E2
    Conclusion: Taken together, PKCθ-E2
    MeSH term(s) Animals ; CD4-Positive T-Lymphocytes/immunology ; HEK293 Cells ; Humans ; Jurkat Cells ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Mutation ; Phenotype ; Protein Kinase C-theta/genetics ; Protein Kinase C-theta/metabolism
    Chemical Substances Protein Kinase C-theta (EC 2.7.11.13)
    Language English
    Publishing date 2019-05-28
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1478-811X
    ISSN (online) 1478-811X
    DOI 10.1186/s12964-019-0364-0
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Loss-of-function phenotype of a PKCθ

    Thuille, Nikolaus / Siegmund, Kerstin / Klepsch, Victoria / Schörgenhuber, Jacqueline / Danklmaier, Sarah / Leitges, Michael / Baier, Gottfried

    Cell communication and signaling : CCS

    2019  Volume 17, Issue 1, Page(s) 141

    Abstract: Background: Protein kinase C θ has been established as an important signaling intermediate in T-effector-cell activation and survival pathways by controlling activity of the key transcription factors NF-κB and NFAT. Previous studies identified an ... ...

    Abstract Background: Protein kinase C θ has been established as an important signaling intermediate in T-effector-cell activation and survival pathways by controlling activity of the key transcription factors NF-κB and NFAT. Previous studies identified an activation-induced auto-phosphorylation site at Thr-219, located between the tandem C1 domains of the regulatory fragment in PKCθ, as a structural requirement for its correct membrane translocation and the subsequent transactivation of downstream signals leading to IL-2 production in a human T cell line.
    Methods: The present work aimed to define the role of this phosphorylation switch on PKCθ in a physiological context through a homozygous T219A knockin mouse strain. T cell activation was analyzed by H3-thymidine uptake (proliferative response), qRT-PCR and luminex measurements (cytokine production). NFAT and NF-κB transactivation responses were estimated by Gel mobility shift and Alpha Screen assays. Frequencies of T cell subsets were analyzed by flow cytometry.
    Results: Despite a normal T cell development, in vitro activated effector T cells clearly revealed a requirement of Thr-219 phosphorylation site on PKCθ for a transactivation of NF-κB and NFAT transcription factors and, subsequently, robust IL-2 and IFN-γ expression.
    Conclusion: This phenotype is reminiscent of the PKCθ knockout T cells, physiologically validating that this (p) Thr-219 auto-phosphorylation site indeed critically regulates PKCθ function in primary mouse T cells.
    MeSH term(s) Animals ; Cytokines/metabolism ; Gene Knock-In Techniques ; Mice ; Phenotype ; Protein Kinase C-theta/genetics ; Protein Kinase C-theta/metabolism ; T-Lymphocytes/cytology ; T-Lymphocytes/metabolism
    Chemical Substances Cytokines ; Protein Kinase C-theta (EC 2.7.11.13)
    Language English
    Publishing date 2019-11-06
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1478-811X
    ISSN (online) 1478-811X
    DOI 10.1186/s12964-019-0466-8
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Addressing the role of PKD3 in the T cell compartment with knockout mice.

    Koutník, Jiří / Neururer, Verena / Gruber, Thomas / Peer, Sebastian / Hermann-Kleiter, Natascha / Olson, William J / Labi, Verena / Leitges, Michael / Baier, Gottfried / Siegmund, Kerstin

    Cell communication and signaling : CCS

    2022  Volume 20, Issue 1, Page(s) 54

    Abstract: Background: The Protein kinase D3 (PKD3) has been implicated in signal transduction downstream of the T cell receptor (TCR). However, its role for the activation of primary T lymphocytes has not been elucidated so far.: Methods: Expression of PKD ... ...

    Abstract Background: The Protein kinase D3 (PKD3) has been implicated in signal transduction downstream of the T cell receptor (TCR). However, its role for the activation of primary T lymphocytes has not been elucidated so far.
    Methods: Expression of PKD isoforms in primary murine T cells was determined by RT-PCR and SDS-Page. A germline PKD3-knockout mouse line was analyzed for its immune response to OVA/alum intraperitoneal immunization. Phenotyping of the T cell compartment ex vivo as well as upon stimulation in vitro was performed by flow cytometry. Additionally, cytokine expression was assessed by flow cytometry, RT-PCR and Luminex technology.
    Results: PKD expression in T cells is modulated by TCR stimulation, leading to a rapid down-regulation on mRNA and on protein level. PKD3-deficient mice respond to immunization with enhanced T follicular helper cell generation. Furthermore, peripheral PKD3-deficient CD4
    Conclusion: While PKD3-deficiency in vivo in mice leads to a skewing of the T cell compartment towards a more activated phenotype, this kinase seems to be dispensable for naïve CD4
    MeSH term(s) Animals ; CD4-Positive T-Lymphocytes ; DNA-Activated Protein Kinase/metabolism ; DNA-Binding Proteins/metabolism ; Mice ; Mice, Knockout ; Receptors, Antigen, T-Cell/metabolism ; T-Lymphocytes/metabolism
    Chemical Substances DNA-Binding Proteins ; Receptors, Antigen, T-Cell ; protein kinase C nu (EC 2.7.1.-) ; DNA-Activated Protein Kinase (EC 2.7.11.1) ; Prkdc protein, mouse (EC 2.7.11.1)
    Language English
    Publishing date 2022-04-19
    Publishing country England
    Document type Journal Article ; Video-Audio Media ; Research Support, Non-U.S. Gov't
    ZDB-ID 2126315-2
    ISSN 1478-811X ; 1478-811X
    ISSN (online) 1478-811X
    ISSN 1478-811X
    DOI 10.1186/s12964-022-00864-w
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Proof of Principle for a T Lymphocyte Intrinsic Function of Coronin 1A.

    Siegmund, Kerstin / Klepsch, Victoria / Hermann-Kleiter, Natascha / Baier, Gottfried

    The Journal of biological chemistry

    2016  Volume 291, Issue 42, Page(s) 22086–22092

    Abstract: Coronins are evolutionarily conserved proteins that were originally identified as modulators of actin-dependent processes. Studies analyzing complete Coronin 1a knock-out mice have shown that this molecule is an important regulator of naive T cell ... ...

    Abstract Coronins are evolutionarily conserved proteins that were originally identified as modulators of actin-dependent processes. Studies analyzing complete Coronin 1a knock-out mice have shown that this molecule is an important regulator of naive T cell homeostasis and it has been linked to immune deficiencies as well as autoimmune disorders. Nevertheless, because Coronin 1A is strongly expressed in all leukocyte subsets, it is not conclusive whether or not this phenotype is attributed to a T cell-intrinsic function of Coronin 1A. To address this research question, we have generated a T cell-specific Coronin 1a knock-out mouse (Coro1a
    MeSH term(s) Animals ; CD4-Positive T-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/pathology ; Encephalomyelitis, Autoimmune, Experimental/genetics ; Encephalomyelitis, Autoimmune, Experimental/immunology ; Encephalomyelitis, Autoimmune, Experimental/pathology ; Immunologic Memory ; Mice ; Mice, Knockout ; Microfilament Proteins/genetics ; Microfilament Proteins/immunology ; Multiple Sclerosis/genetics ; Multiple Sclerosis/immunology ; Multiple Sclerosis/pathology
    Chemical Substances Microfilament Proteins ; coronin proteins (145420-64-0)
    Language English
    Publishing date 2016-08-26
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1074/jbc.M116.748012
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    Uc I Zs.108: Show issues Location:
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