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t cells  (Miltenyi Biotec)


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    Miltenyi Biotec t cells
    T Cells, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pan+t+cell+isolation+kit+ii/Pan+T+Cell+Isolation+Kit+II%2C+mouse/pmc13683377-615-7-11
    Average 97 stars, based on 1190 article reviews
    t cells - by Bioz Stars, 2026-11
    97/100 stars

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    Related Articles

    Isolation:

    Article Title: Neutrophil regulation of immunotherapy for cancer is controlled by type II interferon.
    Article Snippet: .. On the second day, naı̈ve T cells from healthy wildtype mouse spleens were magnetically isolated using the Pan T cell Isolation Kit II (130-095-130, mouse, Miltenyi Biotech). ..

    Article Title: Autophagy regulates the function of myeloid-derived suppressor cells in tumor-induced immunosuppression
    Article Snippet: .. Naïve CD3+ T cells were isolated from the spleens of non-tumor-bearing mice using the Pan T Cell Isolation Kit II (Miltenyi Biotec) and labeled with CFSE (Thermo Fisher Scientific). ..

    Article Title: Circulating Platelets and Microvesicles Regulate Splenic T Cell Responses in Murine Atherosclerosis
    Article Snippet: Preparation of single-cell suspensions from mouse spleen without enzymatic treatment was performed using a protocol described by the producers (Jungblut et al., 2008) with the gentleMACS Dissociator (Miltenyi Biotec). .. The separation of T cells from the suspension was performed on ice using the Pan T Cell Isolation Kit II (negative isolation; Order no. 130-095-130, Miltenyi Biotec). .. Briefly, 40 μl of buffer for 1 × 10^7 cells was mixed with 10 μl of Biotin-Antibody Cocktail for 5 minutes at 4oC, followed by 30 μl of buffer for 1 × 10^7 cells and 20 μl of Anti-Biotin MicroBeads for 10 minutes at 4oC.

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity
    Article Snippet: At the endpoint, supernatants were collected for IFN-γ and granzyme B determination and cell proliferation of B16BL6 cells was analyzed via the WST-assay (Roche Diagnostics GmbH, Mannheim, Germany, #11644807001) after detaching the T-cells. .. CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads. .. The cells were plated into cell culture inserts with 5.0 μm pore polycarbonate membrane (Corning, Kaiserslautern, Germany, #3421) into 24-well plates in medium containing 0.1% FCS.

    Article Title: Opposing functions of gut immunomodulatory metabolites on CAR-T therapy.
    Article Snippet: Splenocytes were collected by centrifugation, and red blood cells were lysed using RBC Lysis Buffer (iNtRON Biotechnology, South Korea) for 2 minutes at room temperature. .. Pan T cells were isolated using the Pan T Cell Isolation Kit II (Miltenyi Biotec, Germany) following the manufacturer’s protocol. ..

    Article Title: A high-molecular-weight polysaccharide from Polygonatum sibiricum inhibits distant tumor growth associated with gut microbiota remodeling and synergizes with αPD-1 therapy.
    Article Snippet: Background: Defined polysaccharide fractions can reshape the gut microbiome and influence systemic antitumor immunity.. We investigated whether an operationally defined high-molecular-weight Polygonatum sibiricum polysaccharide fraction (PSP-H) enriched by 100 kDa ultrafiltration suppresses growth of subcutaneous MC38 tumors via microbiota-dependent mechanisms and potentiates anti-PD-1 therapy.. Materials and methods: PSP-H was isolated by cascade ultrafiltration and compared with a total polysaccharide extract (PSP-T) and lower-MW fractions.

    Cell Isolation:

    Article Title: Neutrophil regulation of immunotherapy for cancer is controlled by type II interferon.
    Article Snippet: .. On the second day, naı̈ve T cells from healthy wildtype mouse spleens were magnetically isolated using the Pan T cell Isolation Kit II (130-095-130, mouse, Miltenyi Biotech). ..

    Article Title: Autophagy regulates the function of myeloid-derived suppressor cells in tumor-induced immunosuppression
    Article Snippet: .. Naïve CD3+ T cells were isolated from the spleens of non-tumor-bearing mice using the Pan T Cell Isolation Kit II (Miltenyi Biotec) and labeled with CFSE (Thermo Fisher Scientific). ..

    Article Title: Circulating Platelets and Microvesicles Regulate Splenic T Cell Responses in Murine Atherosclerosis
    Article Snippet: Preparation of single-cell suspensions from mouse spleen without enzymatic treatment was performed using a protocol described by the producers (Jungblut et al., 2008) with the gentleMACS Dissociator (Miltenyi Biotec). .. The separation of T cells from the suspension was performed on ice using the Pan T Cell Isolation Kit II (negative isolation; Order no. 130-095-130, Miltenyi Biotec). .. Briefly, 40 μl of buffer for 1 × 10^7 cells was mixed with 10 μl of Biotin-Antibody Cocktail for 5 minutes at 4oC, followed by 30 μl of buffer for 1 × 10^7 cells and 20 μl of Anti-Biotin MicroBeads for 10 minutes at 4oC.

    Article Title: Armored Chimeric Antigen Receptor-T Cell Therapy Targets Antigen-Heterogeneous Glioma
    Article Snippet: Next, red blood cells were lysed with 1x Red Blood Cell Lysis Solution (Miltenyi, Cat# 130-094-183). .. The remaining splenocytes were processed via magnetic-bead negative selection to isolate pan-T cells, using a Pan T Cell Isolation Kit II (Miltenyi, Cat# 130-095-130). .. The isolated pan-T cells were then activated using Mouse Dynabead T-cell Activator (ThermoFisher, Cat# 11453D) beads at a 1:1 cell to bead ratio, and cultured in RPMI 1640 with HEPES supplemented with 10% HI-FBS + 20 mM HEPES + 1 mM Sodium Pyruvate (ThermoFisher, Cat# 11360070) + 1 mL GlutaMAX TM (ThermoFisher, Cat# 35050061) + 1x Antibiotic/Antimycotic (Cytiva, Cat# SV30079.01) + 55 μM 2-Mercaptoethanol (ThermoFisher, Cat# 21985023) + 40 U/mL human IL-2 (Miltenyi, Cat# 130-097-748).

    Article Title: Opposing functions of gut immunomodulatory metabolites on CAR-T therapy.
    Article Snippet: Splenocytes were collected by centrifugation, and red blood cells were lysed using RBC Lysis Buffer (iNtRON Biotechnology, South Korea) for 2 minutes at room temperature. .. Pan T cells were isolated using the Pan T Cell Isolation Kit II (Miltenyi Biotec, Germany) following the manufacturer’s protocol. ..

    Article Title: A high-molecular-weight polysaccharide from Polygonatum sibiricum inhibits distant tumor growth associated with gut microbiota remodeling and synergizes with αPD-1 therapy.
    Article Snippet: Background: Defined polysaccharide fractions can reshape the gut microbiome and influence systemic antitumor immunity.. We investigated whether an operationally defined high-molecular-weight Polygonatum sibiricum polysaccharide fraction (PSP-H) enriched by 100 kDa ultrafiltration suppresses growth of subcutaneous MC38 tumors via microbiota-dependent mechanisms and potentiates anti-PD-1 therapy.. Materials and methods: PSP-H was isolated by cascade ultrafiltration and compared with a total polysaccharide extract (PSP-T) and lower-MW fractions.

    Article Title: CAR T cell targeting of inflammatory myeloid progenitors in the bone marrow remodels border-associated macrophages and reverses cognitive aging
    Article Snippet: .. Following tissue dissection and red blood cell lysis using ACK Lysing Buffer (Thermo Fisher, A1049201), primary mouse T cells were purified using the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec, 130-095-130). .. Purified T cells were cultured in RPMI 1640 Medium (Fisher Scientific, 11-875-093) supplemented with 10% FBS (Corning, 35-010-CV), 10 mM HEPES (Thermo Fisher, 15630106), 2 mM L-glutamine (Thermo Fisher, 25030081), 1× MEM Non-Essential Amino Acids Solution (Fisher Scientific, 11-140-050), 55 μM 2-mercaptoethanol (Thermo Fisher, 31350010), 1 mM sodium pyruvate (Thermo Fisher, 11360070), 200 IU/mL recombinant mouse IL-2 (PeproTech, 212-12-500UG), and Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11452D) at a bead:cell ratio of 1:2.

    Labeling:

    Article Title: Autophagy regulates the function of myeloid-derived suppressor cells in tumor-induced immunosuppression
    Article Snippet: .. Naïve CD3+ T cells were isolated from the spleens of non-tumor-bearing mice using the Pan T Cell Isolation Kit II (Miltenyi Biotec) and labeled with CFSE (Thermo Fisher Scientific). ..

    Suspension:

    Article Title: Circulating Platelets and Microvesicles Regulate Splenic T Cell Responses in Murine Atherosclerosis
    Article Snippet: Preparation of single-cell suspensions from mouse spleen without enzymatic treatment was performed using a protocol described by the producers (Jungblut et al., 2008) with the gentleMACS Dissociator (Miltenyi Biotec). .. The separation of T cells from the suspension was performed on ice using the Pan T Cell Isolation Kit II (negative isolation; Order no. 130-095-130, Miltenyi Biotec). .. Briefly, 40 μl of buffer for 1 × 10^7 cells was mixed with 10 μl of Biotin-Antibody Cocktail for 5 minutes at 4oC, followed by 30 μl of buffer for 1 × 10^7 cells and 20 μl of Anti-Biotin MicroBeads for 10 minutes at 4oC.

    Gene Knockout:

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity
    Article Snippet: At the endpoint, supernatants were collected for IFN-γ and granzyme B determination and cell proliferation of B16BL6 cells was analyzed via the WST-assay (Roche Diagnostics GmbH, Mannheim, Germany, #11644807001) after detaching the T-cells. .. CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads. .. The cells were plated into cell culture inserts with 5.0 μm pore polycarbonate membrane (Corning, Kaiserslautern, Germany, #3421) into 24-well plates in medium containing 0.1% FCS.

    Activation Assay:

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity
    Article Snippet: At the endpoint, supernatants were collected for IFN-γ and granzyme B determination and cell proliferation of B16BL6 cells was analyzed via the WST-assay (Roche Diagnostics GmbH, Mannheim, Germany, #11644807001) after detaching the T-cells. .. CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads. .. The cells were plated into cell culture inserts with 5.0 μm pore polycarbonate membrane (Corning, Kaiserslautern, Germany, #3421) into 24-well plates in medium containing 0.1% FCS.

    Selection:

    Article Title: Armored Chimeric Antigen Receptor-T Cell Therapy Targets Antigen-Heterogeneous Glioma
    Article Snippet: Next, red blood cells were lysed with 1x Red Blood Cell Lysis Solution (Miltenyi, Cat# 130-094-183). .. The remaining splenocytes were processed via magnetic-bead negative selection to isolate pan-T cells, using a Pan T Cell Isolation Kit II (Miltenyi, Cat# 130-095-130). .. The isolated pan-T cells were then activated using Mouse Dynabead T-cell Activator (ThermoFisher, Cat# 11453D) beads at a 1:1 cell to bead ratio, and cultured in RPMI 1640 with HEPES supplemented with 10% HI-FBS + 20 mM HEPES + 1 mM Sodium Pyruvate (ThermoFisher, Cat# 11360070) + 1 mL GlutaMAX TM (ThermoFisher, Cat# 35050061) + 1x Antibiotic/Antimycotic (Cytiva, Cat# SV30079.01) + 55 μM 2-Mercaptoethanol (ThermoFisher, Cat# 21985023) + 40 U/mL human IL-2 (Miltenyi, Cat# 130-097-748).

    Tissue:

    Article Title: CAR T cell targeting of inflammatory myeloid progenitors in the bone marrow remodels border-associated macrophages and reverses cognitive aging
    Article Snippet: .. Following tissue dissection and red blood cell lysis using ACK Lysing Buffer (Thermo Fisher, A1049201), primary mouse T cells were purified using the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec, 130-095-130). .. Purified T cells were cultured in RPMI 1640 Medium (Fisher Scientific, 11-875-093) supplemented with 10% FBS (Corning, 35-010-CV), 10 mM HEPES (Thermo Fisher, 15630106), 2 mM L-glutamine (Thermo Fisher, 25030081), 1× MEM Non-Essential Amino Acids Solution (Fisher Scientific, 11-140-050), 55 μM 2-mercaptoethanol (Thermo Fisher, 31350010), 1 mM sodium pyruvate (Thermo Fisher, 11360070), 200 IU/mL recombinant mouse IL-2 (PeproTech, 212-12-500UG), and Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11452D) at a bead:cell ratio of 1:2.

    Dissection:

    Article Title: CAR T cell targeting of inflammatory myeloid progenitors in the bone marrow remodels border-associated macrophages and reverses cognitive aging
    Article Snippet: .. Following tissue dissection and red blood cell lysis using ACK Lysing Buffer (Thermo Fisher, A1049201), primary mouse T cells were purified using the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec, 130-095-130). .. Purified T cells were cultured in RPMI 1640 Medium (Fisher Scientific, 11-875-093) supplemented with 10% FBS (Corning, 35-010-CV), 10 mM HEPES (Thermo Fisher, 15630106), 2 mM L-glutamine (Thermo Fisher, 25030081), 1× MEM Non-Essential Amino Acids Solution (Fisher Scientific, 11-140-050), 55 μM 2-mercaptoethanol (Thermo Fisher, 31350010), 1 mM sodium pyruvate (Thermo Fisher, 11360070), 200 IU/mL recombinant mouse IL-2 (PeproTech, 212-12-500UG), and Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11452D) at a bead:cell ratio of 1:2.

    Red Blood Cell Lysis:

    Article Title: CAR T cell targeting of inflammatory myeloid progenitors in the bone marrow remodels border-associated macrophages and reverses cognitive aging
    Article Snippet: .. Following tissue dissection and red blood cell lysis using ACK Lysing Buffer (Thermo Fisher, A1049201), primary mouse T cells were purified using the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec, 130-095-130). .. Purified T cells were cultured in RPMI 1640 Medium (Fisher Scientific, 11-875-093) supplemented with 10% FBS (Corning, 35-010-CV), 10 mM HEPES (Thermo Fisher, 15630106), 2 mM L-glutamine (Thermo Fisher, 25030081), 1× MEM Non-Essential Amino Acids Solution (Fisher Scientific, 11-140-050), 55 μM 2-mercaptoethanol (Thermo Fisher, 31350010), 1 mM sodium pyruvate (Thermo Fisher, 11360070), 200 IU/mL recombinant mouse IL-2 (PeproTech, 212-12-500UG), and Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11452D) at a bead:cell ratio of 1:2.

    Purification:

    Article Title: CAR T cell targeting of inflammatory myeloid progenitors in the bone marrow remodels border-associated macrophages and reverses cognitive aging
    Article Snippet: .. Following tissue dissection and red blood cell lysis using ACK Lysing Buffer (Thermo Fisher, A1049201), primary mouse T cells were purified using the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec, 130-095-130). .. Purified T cells were cultured in RPMI 1640 Medium (Fisher Scientific, 11-875-093) supplemented with 10% FBS (Corning, 35-010-CV), 10 mM HEPES (Thermo Fisher, 15630106), 2 mM L-glutamine (Thermo Fisher, 25030081), 1× MEM Non-Essential Amino Acids Solution (Fisher Scientific, 11-140-050), 55 μM 2-mercaptoethanol (Thermo Fisher, 31350010), 1 mM sodium pyruvate (Thermo Fisher, 11360070), 200 IU/mL recombinant mouse IL-2 (PeproTech, 212-12-500UG), and Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11452D) at a bead:cell ratio of 1:2.



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    Image Search Results


    Tumor growth after depletion of CD8⁺ T-cells. A CD8⁺ T-cells were depleted in wildtype and IKKε KO mice by intraperitoneal injection of anti-mouse CD8α as indicated. IgG injection served as isotype control. B Successful depletion was verified by FACS analysis of spleen and tumor samples. The plot shows a representative example depicting CD4, CD8, and double-negative (DN; CD3⁺, CD4⁻, CD8⁻) T-cell populations. C Tumor growth kinetics for each individual mouse over a 21-day period. D Left panel: tumor growth kinetics including only mice that survived at least until day 16. Two-way ANOVA with Tukey’s multiple-comparisons test was applied. Arrows indicate tumor volumes from isotype-control–treated mice. Right panel: tumor mass, analyzed by one-way ANOVA with Tukey’s post hoc test. Group sizes: all mice: WT + isotype control, n = 6; WT + anti-CD8α, n = 6; IKKε-KO + isotype control, n = 5; IKKε-KO + anti-CD8α, n = 6.; mice that survived until day 16: WT + isotype control, n = 5; WT + anti-CD8α, n = 3 mice; IKKε-KO + isotype control, n = 4; IKKε-KO + anti-CD8α, n = 5

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Tumor growth after depletion of CD8⁺ T-cells. A CD8⁺ T-cells were depleted in wildtype and IKKε KO mice by intraperitoneal injection of anti-mouse CD8α as indicated. IgG injection served as isotype control. B Successful depletion was verified by FACS analysis of spleen and tumor samples. The plot shows a representative example depicting CD4, CD8, and double-negative (DN; CD3⁺, CD4⁻, CD8⁻) T-cell populations. C Tumor growth kinetics for each individual mouse over a 21-day period. D Left panel: tumor growth kinetics including only mice that survived at least until day 16. Two-way ANOVA with Tukey’s multiple-comparisons test was applied. Arrows indicate tumor volumes from isotype-control–treated mice. Right panel: tumor mass, analyzed by one-way ANOVA with Tukey’s post hoc test. Group sizes: all mice: WT + isotype control, n = 6; WT + anti-CD8α, n = 6; IKKε-KO + isotype control, n = 5; IKKε-KO + anti-CD8α, n = 6.; mice that survived until day 16: WT + isotype control, n = 5; WT + anti-CD8α, n = 3 mice; IKKε-KO + isotype control, n = 4; IKKε-KO + anti-CD8α, n = 5

    Article Snippet: CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads.

    Techniques: Injection, Control, Analysis

    Epidemiology and expression of IKKε in human skin cancer. A Survival proportions of skin cancer patients in relation to IKKε expression levels, adapted from the Human Protein Atlas ( https://www.proteinatlas.org/).The cut-off IKKε expression was calculated by X-tile optimization. * P < 0.05, significant difference between low and high IKKε expression. B Multiplex immunofluorescence analysis showing protein expression of IKKε (red), tumor (S100, blue) and T-cells (CD3 + , green; CD4 + , yellow; CD8⁺, turquoise; γδ-T-cells, orange) in human primary melanoma (representative picture from n = 9 patient samples), Scale Bars: 50 µm. C Quantitative analysis of tumor cells (S100⁺) and CD3⁺ T cells, including colocalization of IKKε expression within tumor cells and CD3⁺ T cells. Values are presented as the percentage of S100⁺ or CD3⁺ cells normalized to the total cell count, or as the proportion of IKKε-positive cells relative to the total number of analyzed cells within the cell type (tumor cells or CD3⁺ T cells), respectively. D Quantitative analysis of T-cell subsets in human melanoma samples. Values are shown as percentages relative to the CD3⁺ T-cell population. Horizontal lines indicate the median. Each dot represents one patient sample ( n = 9). For each sample, 3–8 representative fields of view were analyzed

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Epidemiology and expression of IKKε in human skin cancer. A Survival proportions of skin cancer patients in relation to IKKε expression levels, adapted from the Human Protein Atlas ( https://www.proteinatlas.org/).The cut-off IKKε expression was calculated by X-tile optimization. * P < 0.05, significant difference between low and high IKKε expression. B Multiplex immunofluorescence analysis showing protein expression of IKKε (red), tumor (S100, blue) and T-cells (CD3 + , green; CD4 + , yellow; CD8⁺, turquoise; γδ-T-cells, orange) in human primary melanoma (representative picture from n = 9 patient samples), Scale Bars: 50 µm. C Quantitative analysis of tumor cells (S100⁺) and CD3⁺ T cells, including colocalization of IKKε expression within tumor cells and CD3⁺ T cells. Values are presented as the percentage of S100⁺ or CD3⁺ cells normalized to the total cell count, or as the proportion of IKKε-positive cells relative to the total number of analyzed cells within the cell type (tumor cells or CD3⁺ T cells), respectively. D Quantitative analysis of T-cell subsets in human melanoma samples. Values are shown as percentages relative to the CD3⁺ T-cell population. Horizontal lines indicate the median. Each dot represents one patient sample ( n = 9). For each sample, 3–8 representative fields of view were analyzed

    Article Snippet: CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads.

    Techniques: Expressing, Multiplex Assay, Immunofluorescence, Analysis, Cell Characterization

    Tumor growth, immune cell composition and CD3 T-cell immunofluorescence. A Tumor growth in wildtype (WT) and IKKε KO mice over a period of 21 days and tumor masses at the end of the experiment. WT: n = 13 mice, IKKε KO: n = 15 mice, Two-way Anova, Tukey´s multiple comparisons test; tumor mass, Mann Whitney test. B Immune cell numbers in the tumors as determined by flow cytometry. Cell counts were normalized to the number of living CD45 + immune cells. n = 6/group; Student´s t-test. C Immunofluorescence analysis of CD3-positive cells in the tumor. Nuclei were stained with DAPI and CD3 signals normalized to the cell number. Scale Bar represents 100 µm. Representative figure from WT n = 7, and IKKε KO n = 6, the diagram shows the quantitative analysis of all tumors analyzed. Student´s t-test. * P < 0.05, ** P < 0.01, *** P < 0.001

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Tumor growth, immune cell composition and CD3 T-cell immunofluorescence. A Tumor growth in wildtype (WT) and IKKε KO mice over a period of 21 days and tumor masses at the end of the experiment. WT: n = 13 mice, IKKε KO: n = 15 mice, Two-way Anova, Tukey´s multiple comparisons test; tumor mass, Mann Whitney test. B Immune cell numbers in the tumors as determined by flow cytometry. Cell counts were normalized to the number of living CD45 + immune cells. n = 6/group; Student´s t-test. C Immunofluorescence analysis of CD3-positive cells in the tumor. Nuclei were stained with DAPI and CD3 signals normalized to the cell number. Scale Bar represents 100 µm. Representative figure from WT n = 7, and IKKε KO n = 6, the diagram shows the quantitative analysis of all tumors analyzed. Student´s t-test. * P < 0.05, ** P < 0.01, *** P < 0.001

    Article Snippet: CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads.

    Techniques: Immune Cell, Immunofluorescence, MANN-WHITNEY, Flow Cytometry, Analysis, Staining

    T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

    Article Snippet: CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads.

    Techniques: Migration, Activity Assay, Isolation, Cell Culture, Analysis, Counting, Co-culture Assay, Derivative Assay, WST Assay, Comparison, Enzyme-linked Immunosorbent Assay, Knock-Out

    Proteomic analysis of B16BL6 cells with and without co-culture with WT or IKKε KO CD8⁺ T-cells. B16BL6 were either cultured as monoculture or as co-culture with CD8⁺ T-cells from wildtype and IKKε KO mice. Resulting protein lysates were subjected to proteome analysis by LC-MSMS. A Venn diagram, PCA and Volcano plots showing apparent differences between mono- and co-culture as well as proteins regulated by wildtype or IKKε KO T-cells, respectively. B Gene ontology analysis of differentially regulated proteins. Proteins involved in the significant regulation of the inflammatory response in co-culture of B16BL6 with IKKε KO T-cells are indicated in the respective Volcano plot. n = 5 B16BL6 monoculture-and n = 4 for B16BL6-T-cell co-cultures

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Proteomic analysis of B16BL6 cells with and without co-culture with WT or IKKε KO CD8⁺ T-cells. B16BL6 were either cultured as monoculture or as co-culture with CD8⁺ T-cells from wildtype and IKKε KO mice. Resulting protein lysates were subjected to proteome analysis by LC-MSMS. A Venn diagram, PCA and Volcano plots showing apparent differences between mono- and co-culture as well as proteins regulated by wildtype or IKKε KO T-cells, respectively. B Gene ontology analysis of differentially regulated proteins. Proteins involved in the significant regulation of the inflammatory response in co-culture of B16BL6 with IKKε KO T-cells are indicated in the respective Volcano plot. n = 5 B16BL6 monoculture-and n = 4 for B16BL6-T-cell co-cultures

    Article Snippet: CD3 T-cells were isolated from the spleen of wildtype and IKKε KO mice using a Pan T-cell Isolation Kit II, mouse, (Miltenyi Bergisch Gladbach, Germany, #130–095–130) and directly used without prior activation with CD3/CD28 Beads.

    Techniques: Analysis, Co-culture Assay, Cell Culture, Co-Cultures