Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: NK cell-triggered CCL5/IFNγ-CXCL9/10 axis underlies the clinical efficacy of neoadjuvant anti-HER2 antibodies in breast cancer

doi: 10.1186/s13046-023-02918-4

Figure Lengend Snippet: Specialized tumor-infiltrating NK cell subsets associate to distinct immune contextures in human breast tumors. Unsorted multicellular suspensions derived from fresh breast tumor specimens were cultured in complete medium with IL2 in the presence or absence of trastuzumab (Tz; 210 ng/ml) for 24 h. A Dot plots showing the NK cell gate (CD56 + CD3 - ) used for the analysis of the expression of CD16 and CD103 in TI-NK cells of three representative samples. B-C Proportions of CD137 + cells in CD16 + , CD16 - CD103 + and CD16 - CD103 - subpopulations in fresh breast tumor-derived multicellular cultures treated or not with trastuzumab. D-G Fresh breast tumor specimens ( n = 84) were processed and stained with a combination of antibodies specific for CD45, CD3, CD56, CD8, CD4, CD16, CD103, NKG2C and PD1 and analysed by flow cytometry. D tSNE of major alive lymphocyte subsets (DAPI - CD45 + ) in breast tumor immune infiltrates generated with data from 18 tumors. Red, blue and green circles indicate major NK, CD4 and CD8 T cell subsets, respectively. E Percentage of CD16 + , CD16 - CD103 + and CD16 - CD103 - TI-NK cell subpopulations in treatment naïve, fresh tumor samples. F , G Heat maps showing Spearman’s correlation coefficients between the indicated lymphocyte subsets in breast tumors ( n = 73–84) and HER2 tumors ( n = 20). Asterisks label significant correlations (**** p < 0.0001; *** p < 0.001; ** p < 0.01; * p < 0.05

Article Snippet: After culture, cells were stained with a combination of directly labeled antibodies, which included: anti-CD45 (clone 2D1, 56–9459-42, eBiosciences), anti-CD56 (clone CMSSB, 17–0567-42, Invitrogen), anti-CD3 (clone SK7, 345766, BD), anti-CD16 (clone CB16, 47–0168-42, eBiosciences), anti-CD103 (clone B-Ly7, 11–1038-42, eBiosciences) and anti-CD137 (clone 4B4, 12–1379, eBiosciences).

Techniques: Derivative Assay, Cell Culture, Expressing, Staining, Flow Cytometry, Generated

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L signaling activates microglia in vitro . Microglia cells were grown on plates that had been coated with PBS (grey bars) or 10 μg/mL of Fc control protein (white bars) or 10 μg/mL of CD137-Fc protein (black bars). ( A ) Attachment and morphological changes of primary microglia were documented by photography (40× magnification) 1 h after plating. Scale bar: 20 μm. ( B ) Attachment and morphological changes of BV-2 and N9 cells and primary microglia were documented by photography (63×) 24 h after plating. Cells exposed to immobilized CD137-Fc protein developed long spiky projections* and became amoeboid cells^ with shortened protrusions # . Scale bar: 20 μm. ( C ) The concentrations of cytokines in supernatants of primary microglia were determined by ELISA at indicated time points. Depicted are means ± standard deviations of triplicate measurements. ( D ) The phagocytic capacity of the cells was determined by adding FITC-labeled latex beads for 1 h before analysis by flow cytometry. Control: Autofluorescence of the cells. Numbers above the histograms state the percentages of positive cells and mean fluorescence intensities (MFI). These experiments were repeated three times with similar results. * P <0.05; ** P <0.01.

Article Snippet: Unspecific staining was blocked by 2% serum for 30 min. Endogenous peroxidases were inactivated by 3% hydrogen peroxide for 15 min. Anti-CD137 (goat polyclonal, R&D Systems) and anti-Iba-1 (rabbit polyclonal, Wako Chemicals) in PBS were used as primary antibodies and hybridized overnight.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Labeling, Flow Cytometry, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L signaling activates microglia in vivo . ( A ) CD137L is required for activation of microglia in vivo . Cortex and spinal cord tissue sections of WT and CD137L -/- mice with EAE were immunohistochemically stained with an isotype control antibody or for Iba-1 (brown). Shown in the inset is a close-up of a single Iba-1-positive microglia cell in the cortex of a WT mouse with EAE. ( B ) Quantification of Iba-1 + microglia in the spinal cords and cortices of WT and CD137L -/- mice with EAE. Evaluated were three fields from two sections each using the Metamorph NX Software. Depicted are means ± standard deviations. ( C ) CD137 is expressed in the CNS during EAE. Spinal cord of naïve WT mice and WT mice with EAE was sectioned and stained with an isotype control antibody or for CD137 (brown). ( D ) The presence of CD137L is required for oligodendrocyte apoptosis in EAE. Tissue sections from the dorsal column of the spinal cord and the white matter of the cerebellum of WT and CD137L -/- mice with EAE were stained for oligodendrocytes using a Cy3-labeled anti-Nogo-A antibody (red). Apoptosis was detected by TUNEL staining (green). Nuclei were visualized by DAPI (blue). The yellow staining results from an overlay of red and green and indicates apoptotic oligodendrocytes. Magnification: 40×. ( E ) Quantification of apoptotic oligodendrocytes in the cerebellum of WT and CD137L -/- mice with EAE. Evaluated were three fields from two sections each using the Metamorph NX Software. Depicted are means ± standard deviations.

Article Snippet: Unspecific staining was blocked by 2% serum for 30 min. Endogenous peroxidases were inactivated by 3% hydrogen peroxide for 15 min. Anti-CD137 (goat polyclonal, R&D Systems) and anti-Iba-1 (rabbit polyclonal, Wako Chemicals) in PBS were used as primary antibodies and hybridized overnight.

Techniques: In Vivo, Activation Assay, Staining, Software, Labeling, TUNEL Assay

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: Induction of oligodendrocyte death by CD137L-activated microglia. (A ) Expression of CD137L on the oligodendrocyte cell line OLN93 was determined by flow cytometry. Open histogram: Isotype control. Grey histogram: Anti-CD137L monoclonal antibody (clone TKS-1). ( B ) N9 and OLN93 cells were cultured for 24 h at a 1:1 ratio (1.5×10 5 each) on plates that had been coated with nothing (PBS) or 10 μg/mL of Fc control protein or 10 μg/mL of CD137-Fc protein. ( C ) The rates of OLN93 cell apoptosis in co-cultures with primary microglia from WT or CD137L -/- mice was determined 48 h after initiation of CD137L signaling by 7-AAD and Annexin V staining. Numbers in quadrants indicate percentages of cells. These experiments were repeated two to three times with similar results. * P <0.05; ** P <0.01.

Article Snippet: Unspecific staining was blocked by 2% serum for 30 min. Endogenous peroxidases were inactivated by 3% hydrogen peroxide for 15 min. Anti-CD137 (goat polyclonal, R&D Systems) and anti-Iba-1 (rabbit polyclonal, Wako Chemicals) in PBS were used as primary antibodies and hybridized overnight.

Techniques: Expressing, Flow Cytometry, Cell Culture, Staining

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L-activated microglia induces oligodendrocyte apoptosis via ROS. ( A ) BV-2 cells were cultured on uncoated plates (PBS) or plates coated with Fc or CD137-Fc protein for 24 h, and were then stimulated with 0.4 μg of PMA for 1 h, stained with DHR123 before production of ROS was quantified by flow cytometry. White histogram: No DHR123. The number in the panel indicates the percentage of positive cells. ( B ) BV-2 cells were co-cultured with OLN93 cells at a 1:1 ratio with or without 10,000 U/mL catalase. The rate of apoptosis of cultures was determined after 24 h by 7-AAD and Annexin V staining. Numbers in quadrants indicate percentages of cells. ( C ) The percentages of 7-AAD + OLN93 cells with and without catalase treatment of B are presented as means ± standard deviations of triplicate measurements. * P < 0.05; ** P < 0.01.

Article Snippet: Unspecific staining was blocked by 2% serum for 30 min. Endogenous peroxidases were inactivated by 3% hydrogen peroxide for 15 min. Anti-CD137 (goat polyclonal, R&D Systems) and anti-Iba-1 (rabbit polyclonal, Wako Chemicals) in PBS were used as primary antibodies and hybridized overnight.

Techniques: Cell Culture, Staining, Flow Cytometry

Journal: Gut Pathogens

Article Title: “ Bifidobacterium longum -reactive T helper cells as marker for intestinal barrier impairment in ICU patients with sepsis”

doi: 10.1186/s13099-025-00770-9

Figure Lengend Snippet: Frequencies of circulating CD4 + and CD4 + CD154 + Th cells in cohort 1 ( A ) Frequencies of CD4 + Th cells after stimulation with control, E. coli , C. albicans or B. longum ( n = 20/10/20/10). ( B ) Frequencies of CD4 + CD154 + Th cells after stimulation with control, E. coli , C. albicans or B. longum ( n = 20/10/20/10). ( C ) Frequencies of CD4 + CD154 + Th cells in sepsis patients after stimulation with control or indicated antigens ( n = 20). Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Subsequently, cells were labeled with anti-CD154 (both cohorts) and anti-CD137 (only cohort 1) biotin antibodies followed by anti-biotin MicroBeads (CD154 & CD137 MicroBead Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and magnetically enriched with MS columns (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Control

Journal: Gut Pathogens

Article Title: “ Bifidobacterium longum -reactive T helper cells as marker for intestinal barrier impairment in ICU patients with sepsis”

doi: 10.1186/s13099-025-00770-9

Figure Lengend Snippet: Cytokine production in antigen-reactive CD4 + CD154 + Th cells in cohort 1 ( A ) Frequencies of IFNγ producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans or B. longum ( n = 20/10/20/10). ( B ) Frequencies of TNF-α producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans or B. longum ( n = 20/10/20/10). ( C ) Frequencies of IL-2 producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans or B. longum ( n = 20/10/20/10). Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Subsequently, cells were labeled with anti-CD154 (both cohorts) and anti-CD137 (only cohort 1) biotin antibodies followed by anti-biotin MicroBeads (CD154 & CD137 MicroBead Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and magnetically enriched with MS columns (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques:

Journal: Gut Pathogens

Article Title: “ Bifidobacterium longum -reactive T helper cells as marker for intestinal barrier impairment in ICU patients with sepsis”

doi: 10.1186/s13099-025-00770-9

Figure Lengend Snippet: Frequencies of circulating CD4 + and CD4 + CD154 + Th cells in cohort 2 ( A ) Frequencies of CD4 + Th cells after stimulation with control, E. coli , C. albicans or B. longum ( n = 10/10/10). ( B ) Frequencies of CD4 + CD154 + Th cells after stimulation with control, E. coli , C. albicans , or B. longum ( n = 10/10/10). Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Subsequently, cells were labeled with anti-CD154 (both cohorts) and anti-CD137 (only cohort 1) biotin antibodies followed by anti-biotin MicroBeads (CD154 & CD137 MicroBead Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and magnetically enriched with MS columns (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Control

Journal: Gut Pathogens

Article Title: “ Bifidobacterium longum -reactive T helper cells as marker for intestinal barrier impairment in ICU patients with sepsis”

doi: 10.1186/s13099-025-00770-9

Figure Lengend Snippet: Cytokine production in antigen-reactive CD4 + CD154 + Th cells in cohort 2. ( A ) Frequencies of IL-17 producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans , or B. longum ( n = 10/10/10). ( B ) Frequencies of IL-4 producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans , or B. longum ( n = 10/10/10). ( C ) Frequencies of IL-10 producing CD4 + CD154 + Th cells after stimulation with E. coli , C. albicans or B. longum ( n = 10/10/10). Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Subsequently, cells were labeled with anti-CD154 (both cohorts) and anti-CD137 (only cohort 1) biotin antibodies followed by anti-biotin MicroBeads (CD154 & CD137 MicroBead Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and magnetically enriched with MS columns (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques:

Journal: Gut Pathogens

Article Title: “ Bifidobacterium longum -reactive T helper cells as marker for intestinal barrier impairment in ICU patients with sepsis”

doi: 10.1186/s13099-025-00770-9

Figure Lengend Snippet: Bifidobacterium longum -reactive gut trophic Th cell signature ( A ) Frequencies of CD4 + CD154 + α4ß7 + Th cells after stimulation with E. coli , C. albicans , or B. longum ( n = 10/10/10). Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001. ( B ) Frequencies of B. longum reactive Th cells in cohort 1 and 2. ( C ) Frequencies of IFNγ- and TNF-α producing B. longum reactive Th cells in cohort 1 and 2. ( D ) Frequencies of gut trophic α4ß7 + -expressing B. longum- reactive Th cells. (B) and (C) Merged data from both cohorts. Color code indicating respective groups

Article Snippet: Subsequently, cells were labeled with anti-CD154 (both cohorts) and anti-CD137 (only cohort 1) biotin antibodies followed by anti-biotin MicroBeads (CD154 & CD137 MicroBead Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and magnetically enriched with MS columns (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Expressing