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  1. Article ; Online: The IL-10GFP (VeRT-X) mouse strain is not suitable for the detection of IL-10 production by granulocytes during lung inflammation.

    Özkan, Müge / Eskiocak, Yusuf Cem / Wingender, Gerhard

    PloS one

    2021  Volume 16, Issue 5, Page(s) e0247895

    Abstract: The clear and unequivocal identification of immune effector functions is essential to understand immune responses. The cytokine IL-10 is a critical immune regulator and was shown, for example, to limit pathology during various lung diseases. However, the ...

    Abstract The clear and unequivocal identification of immune effector functions is essential to understand immune responses. The cytokine IL-10 is a critical immune regulator and was shown, for example, to limit pathology during various lung diseases. However, the clear identification of IL-10-producing cells is challenging and, therefore, reporter mouse lines were developed to facilitate their detection. Several such reporter lines utilize GFP, including the IL-10GFP (VeRT-X) reporter strain studied here. In line with previous reports, we found that this IL-10GFP line faithfully reports on the IL-10 production of lymphoid cells. However, we show that the IL-10GFP reporter is not suitable to analyse IL-10 production of myeloid cells during inflammation. During inflammation, the autofluorescence of myeloid cells increased to an extent that entirely masked the IL-10-specific GFP-signal. Our data illustrate a general and important technical caveat using GFP-reporter lines for the analysis of myeloid cells and suggest that previous reports on effector functions of myeloid cells using such GFP-based reporters might require re-evaluation.
    MeSH term(s) Animals ; Flow Cytometry ; Granulocytes/metabolism ; Interleukin-10/metabolism ; Mice ; Mice, Inbred C57BL ; Myeloid Cells/metabolism ; Pneumonia/metabolism
    Chemical Substances Interleukin-10 (130068-27-8)
    Language English
    Publishing date 2021-05-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0247895
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Macrophage and dendritic cell subset composition can distinguish endotypes in adjuvant-induced asthma mouse models.

    Özkan, Müge / Eskiocak, Yusuf Cem / Wingender, Gerhard

    PloS one

    2021  Volume 16, Issue 6, Page(s) e0250533

    Abstract: Asthma is a heterogeneous disease with neutrophilic and eosinophilic asthma as the main endotypes that are distinguished according to the cells recruited to the airways and the related pathology. Eosinophilic asthma is the treatment-responsive endotype, ... ...

    Abstract Asthma is a heterogeneous disease with neutrophilic and eosinophilic asthma as the main endotypes that are distinguished according to the cells recruited to the airways and the related pathology. Eosinophilic asthma is the treatment-responsive endotype, which is mainly associated with allergic asthma. Neutrophilic asthma is a treatment-resistant endotype, affecting 5-10% of asthmatics. Although eosinophilic asthma is well-studied, a clear understanding of the endotypes is essential to devise effective diagnosis and treatment approaches for neutrophilic asthma. To this end, we directly compared adjuvant-induced mouse models of neutrophilic (CFA/OVA) and eosinophilic (Alum/OVA) asthma side-by-side. The immune response in the inflamed lung was analyzed by multi-parametric flow cytometry and immunofluorescence. We found that eosinophilic asthma was characterized by a preferential recruitment of interstitial macrophages and myeloid dendritic cells, whereas in neutrophilic asthma plasmacytoid dendritic cells, exudate macrophages, and GL7+ activated B cells predominated. This differential distribution of macrophage and dendritic cell subsets reveals important aspects of the pathophysiology of asthma and holds the promise to be used as biomarkers to diagnose asthma endotypes.
    MeSH term(s) Animals ; Asthma/immunology ; Dendritic Cells/cytology ; Disease Models, Animal ; Leukocyte Count ; Macrophages/cytology ; Mice ; Neutrophils/immunology
    Language English
    Publishing date 2021-06-01
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0250533
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.

    Eskiocak, Yusuf Cem / Ayyildiz, Zeynep Ozge / Gunalp, Sinem / Korkmaz, Asli / Helvaci, Derya Goksu / Dogan, Yavuz / Sag, Duygu / Wingender, Gerhard

    PloS one

    2023  Volume 18, Issue 2, Page(s) e0282037

    Abstract: Various aspects of the in vitro culture conditions can impact the functional response of immune cells. For example, it was shown that a Ca2+ concentration of at least 1.5 mM during in vitro stimulation is needed for optimal cytokine production by ... ...

    Abstract Various aspects of the in vitro culture conditions can impact the functional response of immune cells. For example, it was shown that a Ca2+ concentration of at least 1.5 mM during in vitro stimulation is needed for optimal cytokine production by conventional αβ T cells. Here we extend these findings by showing that also unconventional T cells (invariant Natural Killer T cells, mucosal-associated invariant T cells, γδ T cells), as well as B cells, show an increased cytokine response following in vitro stimulation in the presence of elevated Ca2+ concentrations. This effect appeared more pronounced with mouse than with human lymphoid cells and did not influence their survival. A similarly increased cytokine response due to elevated Ca2+ levels was observed with primary human monocytes. In contrast, primary human monocyte-derived macrophages, either unpolarized (M0) or polarized into M1 or M2 macrophages, displayed increased cell death in the presence of elevated Ca2+ concentrations. Furthermore, elevated Ca2+ concentrations promoted phenotypic M1 differentiation by increasing M1 markers on M1 and M2 macrophages and decreasing M2 markers on M2 macrophages. However, the cytokine production of macrophages, again in contrast to the lymphoid cells, was unaltered by the Ca2+ concentration. In summary, our data demonstrate that the Ca2+ concentration during in vitro cultures is an important variable to be considered for functional experiments and that elevated Ca2+ levels can boost cytokine production by both mouse and human lymphoid cells.
    MeSH term(s) Humans ; Calcium/metabolism ; Macrophages/metabolism ; Cytokines/metabolism ; Monocytes/metabolism ; Cell Differentiation ; Lymphocytes/metabolism
    Chemical Substances Calcium (SY7Q814VUP) ; Cytokines
    Language English
    Publishing date 2023-02-24
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0282037
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Characterization of a Subset of Patients With Rheumatoid Arthritis for Whom Current Management Strategies are Inadequate.

    Bradford, Claire M / McDonnell, Thomas / Raj, Divya / Robinson, George A / Cole, Andrew / Ramakrishnan, Shashank / González-Serrano, Rosa / Mak, Jasper / Eskiocak, Yusuf Cem / Isenberg, David A / Ciurtin, Coziana / Jury, Elizabeth C / Manson, Jessica J

    ACR open rheumatology

    2019  Volume 1, Issue 3, Page(s) 145–155

    Abstract: Objective: A subset of patients with seropositive rheumatoid arthritis (RA) do not mount a C-reactive protein (CRP) response during flares. We hypothesize that these patients are more likely to experience poor clinical care and less likely to respond to ...

    Abstract Objective: A subset of patients with seropositive rheumatoid arthritis (RA) do not mount a C-reactive protein (CRP) response during flares. We hypothesize that these patients are more likely to experience poor clinical care and less likely to respond to traditional therapy. This study questioned whether this presentation was associated with worse disease outcome and distinct immunological features.
    Methods: Using Power Doppler ultrasound, 48 RA patients with active synovitis were recruited; 30 had normal (n)CRP (5 mg/L or less) and 18 had high (h)CRP (more than 5 mg/L) levels. All had equivalent disease burden assessed by other clinical and laboratory parameters.
    Results: Time to diagnosis and time to first disease-modifying antirheumatic drug were significantly longer in nCRP compared with hCRP patients (
    Conclusion: Patients with nCRP during flares of RA had an altered immunological profile compared with hCRP patients and experienced diagnostic delays and responded less favorably to conventional treatment.
    Language English
    Publishing date 2019-04-22
    Publishing country United States
    Document type Journal Article
    ISSN 2578-5745
    ISSN (online) 2578-5745
    DOI 10.1002/acr2.1021
    Database MEDical Literature Analysis and Retrieval System OnLINE

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