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human cd8 t cells  (Miltenyi Biotec)


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    Miltenyi Biotec human cd8 t cells
    Human Cd8 T Cells, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 373 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd8+t+cell+isolation+kit/CD8%2B+T+Cell+Isolation+Kit%2C+human/us12698265-236-1-14
    Average 97 stars, based on 373 article reviews
    human cd8 t cells - by Bioz Stars, 2026-09
    97/100 stars

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    Related Articles

    Cell Isolation:

    Article Title: A tumor-intrinsic WNT-inhibitory NOTUM program drives immune resistance in microsatellite stable colorectal cancer
    Article Snippet: QIAquick Gel Extraction Kit - Gel Purification , Qiagen , 28704. .. CD8 + T cell Isolation Kit , Miltenyi Biotec , 130-096-495. .. RNAscope Multiplex Fluorescent Reagent Kit v2 , Advanced Cell Diagnostics , 323100.

    Article Title: DC-intrinsic SHP1 restrains central memory CD8⁺ T cell formation via the TCF-1/Wnt/β-Catenin axis.
    Article Snippet: 5 Qingyuan Maternal and Child Healthcare Hospital, No. 22, Shuguang 2Nd Road, Qingcheng District, Qingyuan 511500, China Abstract The tumor microenvironment (TME) limits durable antitumor immunity by impairing CD8+ T cell responses.. Memory like CD8+ T cells are important for long-term immune control but are often restricted in the TME.. Dendritic cells (DCs) are key regulators of T cell fate.

    Article Title: TRIM28-mediated SUMOylation of SREBF1 drives PD‑L1 N‑glycosylation and immune evasion in bladder cancer.
    Article Snippet: Isolation of PBMCs and CD8+ T cells Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors by Ficoll-Paque density gradient centrifugation (GE Healthcare, Cat# 17-5442-02), followed by two PBS washes to remove platelets and impurities. .. CD8+ T cells were purified using a CD8+ T Cell Isolation Kit (Miltenyi Biotec, Cat# 130-096-495). ..

    Article Title: PRKN mediates the ubiquitination of SCAF8 to reduce the mRNA stability of KLF5 and its transcriptional activation of EFNA3 in colorectal cancer.
    Article Snippet: .. Detection of the killing effects of cytotoxic lymphocytes Cytotoxic lymphocytes, including CD8+ T cells and NK cells, were isolated from healthy PBMCs using the CD8+ T Cell Isolation Kit (130-096-495, Miltenyi biotec, Bergisch Gladbach, Germany) AR TIC LE IN PR ES S and NK Cell Isolation Kit (130-092-657, Miltenyi Biotec), respectively. ..

    Article Title: Complement factor B deficiency promotes lung cancer progression via FOS-mediated tryptophan metabolism and immune suppression
    Article Snippet: Absorbance was measured using a microplate reader (BioTek, Winooski, VT, USA; model Synergy H1). .. CD4+CD25+ regulatory T cells and CD8+ T cells were isolated from spleens of 6–8 week-old C57BL/6 mice using the CD4+CD25+ Regulatory T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany; catalog #130-091-041) and CD8+ T Cell Isolation Kit (Miltenyi Biotec, catalog #130-104-075), respectively. ..

    Article Title: FTL008.16, a 5T4-Conditional 4-1BB bispecific antibody, potently enhances antitumor immunity via tumor-directed t-cell activation
    Article Snippet: Human T cells were isolated through immunomagnetic sorting (Miltenyi, #130-097-043) using CD3 antibody-conjugated magnetic beads. .. CD8 + T cells were purified from PBMCs by negative selection using a commercial CD8 + T Cell Isolation Kit (Miltenyi, #130-096-495). .. NK cells were similarly isolated through negative selection (NK Cell Isolation Kit, Miltenyi, #130-092-657) and then pre-stimulated with 10 ng/mL of human IL-2 (Acro, #GMP-L02H14) overnight at 37 °C.

    Article Title: Multimodal sequencing identifies synergistic mechanisms driving resistance to neoadjuvant nivolumab treatment in hepatocellular carcinoma.
    Article Snippet: Peripheral blood was obtained from healthy donors and peripheral blood mononuclear cells (PBMCs) were extracted by Human Lymphocyte Separation Medium (7111011, DAKEWE). .. CD8+ T cells were isolated from human PBMCs using CD8+ T Cell Isolation Kit (130-096-495, Miltenyi Biotec) according to the manufacturer’s instructions and grown in vitro with recombinant IL2 (200-02, PeproTech) and CD3/CD28 Dynabeads (11161D, Thermo Fisher Scientific). ..

    Article Title: Dihydroartemisinin inhibits mutant KRAS to potentiate regorafenib plus anti-PD-1 in KRAS-mutant colorectal cancer liver metastases.
    Article Snippet: CD8+ T cells isolation, expansion and co-culture with cancer cells Human peripheral blood lymphocytes (PBLCs) were isolated with separation medium (LTS 1077, tbdscience, Tianjin, China). .. CD8+ T cells were sorted from PBLCs with the CD8+ T Cell Isolation Kit (130-096-495, Miltenyi Biotech, Germany). .. CD8+ T cells were cultured and expanded in ImmunoCultTM-XF T Cell Expansion Medium (10981, STEMCELL Technologies, Canada) with ImmunoCultTM Human CD3/CD28 T cell activator (10991, STEMCELL Technologies, Canada) and human interleukin (IL) -2 (100 IU/mL, Beijing Four Rings Bio-Pharmaceutical).

    Isolation:

    Article Title: DC-intrinsic SHP1 restrains central memory CD8⁺ T cell formation via the TCF-1/Wnt/β-Catenin axis.
    Article Snippet: 5 Qingyuan Maternal and Child Healthcare Hospital, No. 22, Shuguang 2Nd Road, Qingcheng District, Qingyuan 511500, China Abstract The tumor microenvironment (TME) limits durable antitumor immunity by impairing CD8+ T cell responses.. Memory like CD8+ T cells are important for long-term immune control but are often restricted in the TME.. Dendritic cells (DCs) are key regulators of T cell fate.

    Article Title: PRKN mediates the ubiquitination of SCAF8 to reduce the mRNA stability of KLF5 and its transcriptional activation of EFNA3 in colorectal cancer.
    Article Snippet: .. Detection of the killing effects of cytotoxic lymphocytes Cytotoxic lymphocytes, including CD8+ T cells and NK cells, were isolated from healthy PBMCs using the CD8+ T Cell Isolation Kit (130-096-495, Miltenyi biotec, Bergisch Gladbach, Germany) AR TIC LE IN PR ES S and NK Cell Isolation Kit (130-092-657, Miltenyi Biotec), respectively. ..

    Article Title: Complement factor B deficiency promotes lung cancer progression via FOS-mediated tryptophan metabolism and immune suppression
    Article Snippet: Absorbance was measured using a microplate reader (BioTek, Winooski, VT, USA; model Synergy H1). .. CD4+CD25+ regulatory T cells and CD8+ T cells were isolated from spleens of 6–8 week-old C57BL/6 mice using the CD4+CD25+ Regulatory T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany; catalog #130-091-041) and CD8+ T Cell Isolation Kit (Miltenyi Biotec, catalog #130-104-075), respectively. ..

    Article Title: Multimodal sequencing identifies synergistic mechanisms driving resistance to neoadjuvant nivolumab treatment in hepatocellular carcinoma.
    Article Snippet: Peripheral blood was obtained from healthy donors and peripheral blood mononuclear cells (PBMCs) were extracted by Human Lymphocyte Separation Medium (7111011, DAKEWE). .. CD8+ T cells were isolated from human PBMCs using CD8+ T Cell Isolation Kit (130-096-495, Miltenyi Biotec) according to the manufacturer’s instructions and grown in vitro with recombinant IL2 (200-02, PeproTech) and CD3/CD28 Dynabeads (11161D, Thermo Fisher Scientific). ..

    Purification:

    Article Title: TRIM28-mediated SUMOylation of SREBF1 drives PD‑L1 N‑glycosylation and immune evasion in bladder cancer.
    Article Snippet: Isolation of PBMCs and CD8+ T cells Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors by Ficoll-Paque density gradient centrifugation (GE Healthcare, Cat# 17-5442-02), followed by two PBS washes to remove platelets and impurities. .. CD8+ T cells were purified using a CD8+ T Cell Isolation Kit (Miltenyi Biotec, Cat# 130-096-495). ..

    Article Title: FTL008.16, a 5T4-Conditional 4-1BB bispecific antibody, potently enhances antitumor immunity via tumor-directed t-cell activation
    Article Snippet: Human T cells were isolated through immunomagnetic sorting (Miltenyi, #130-097-043) using CD3 antibody-conjugated magnetic beads. .. CD8 + T cells were purified from PBMCs by negative selection using a commercial CD8 + T Cell Isolation Kit (Miltenyi, #130-096-495). .. NK cells were similarly isolated through negative selection (NK Cell Isolation Kit, Miltenyi, #130-092-657) and then pre-stimulated with 10 ng/mL of human IL-2 (Acro, #GMP-L02H14) overnight at 37 °C.

    Selection:

    Article Title: FTL008.16, a 5T4-Conditional 4-1BB bispecific antibody, potently enhances antitumor immunity via tumor-directed t-cell activation
    Article Snippet: Human T cells were isolated through immunomagnetic sorting (Miltenyi, #130-097-043) using CD3 antibody-conjugated magnetic beads. .. CD8 + T cells were purified from PBMCs by negative selection using a commercial CD8 + T Cell Isolation Kit (Miltenyi, #130-096-495). .. NK cells were similarly isolated through negative selection (NK Cell Isolation Kit, Miltenyi, #130-092-657) and then pre-stimulated with 10 ng/mL of human IL-2 (Acro, #GMP-L02H14) overnight at 37 °C.

    In Vitro:

    Article Title: Multimodal sequencing identifies synergistic mechanisms driving resistance to neoadjuvant nivolumab treatment in hepatocellular carcinoma.
    Article Snippet: Peripheral blood was obtained from healthy donors and peripheral blood mononuclear cells (PBMCs) were extracted by Human Lymphocyte Separation Medium (7111011, DAKEWE). .. CD8+ T cells were isolated from human PBMCs using CD8+ T Cell Isolation Kit (130-096-495, Miltenyi Biotec) according to the manufacturer’s instructions and grown in vitro with recombinant IL2 (200-02, PeproTech) and CD3/CD28 Dynabeads (11161D, Thermo Fisher Scientific). ..

    Recombinant:

    Article Title: Multimodal sequencing identifies synergistic mechanisms driving resistance to neoadjuvant nivolumab treatment in hepatocellular carcinoma.
    Article Snippet: Peripheral blood was obtained from healthy donors and peripheral blood mononuclear cells (PBMCs) were extracted by Human Lymphocyte Separation Medium (7111011, DAKEWE). .. CD8+ T cells were isolated from human PBMCs using CD8+ T Cell Isolation Kit (130-096-495, Miltenyi Biotec) according to the manufacturer’s instructions and grown in vitro with recombinant IL2 (200-02, PeproTech) and CD3/CD28 Dynabeads (11161D, Thermo Fisher Scientific). ..



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    Image Search Results


    GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl CD4 Cre mice. (A) Schematic diagram of the effect of pH on T cell GPCR68 as well as tumor. (B) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with varying pH. RT-qPCR was performed to determine the expression of GPCR68 at various pH. (C) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 under different pH conditions, and GPCR68 protein expression was assessed by Western blot analysis. (D) To generate conditional knockout (CKO) of GPCR68 in T cells, GPCR68 fl/fl mice were crossed with CD4 Cre mice and generated GPCR68 fl/fl CD4 Cre (CKO). (E) Flow cytometry was used to determine the population of CD4 and CD8 cells in the lymph nodes (LN), thymus (THY), and spleen (SP) at the basal level in CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (F) Flow cytometry was used to determine the population of Foxp3+ Treg cells in the lymph nodes, thymus, and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (G-H) The population of F4/80+, CD11c+ (G), and B220+ (H) cells was determined in the lymph nodes and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (I-J) Flow cytometry was used to evaluate the CD4 + or CD8 + T cells for the determination of intracellular cytokines IFN-γ+ (I), or TNF-α+ (J) from the spleen and lymph nodes at basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. Student t-test was performed for comparison between the two groups. Data are mean ± SEM (n = 5), ∗ p < 0.05.

    Journal: Bioactive Materials

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

    doi: 10.1016/j.bioactmat.2026.02.039

    Figure Lengend Snippet: GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl CD4 Cre mice. (A) Schematic diagram of the effect of pH on T cell GPCR68 as well as tumor. (B) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with varying pH. RT-qPCR was performed to determine the expression of GPCR68 at various pH. (C) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 under different pH conditions, and GPCR68 protein expression was assessed by Western blot analysis. (D) To generate conditional knockout (CKO) of GPCR68 in T cells, GPCR68 fl/fl mice were crossed with CD4 Cre mice and generated GPCR68 fl/fl CD4 Cre (CKO). (E) Flow cytometry was used to determine the population of CD4 and CD8 cells in the lymph nodes (LN), thymus (THY), and spleen (SP) at the basal level in CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (F) Flow cytometry was used to determine the population of Foxp3+ Treg cells in the lymph nodes, thymus, and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (G-H) The population of F4/80+, CD11c+ (G), and B220+ (H) cells was determined in the lymph nodes and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (I-J) Flow cytometry was used to evaluate the CD4 + or CD8 + T cells for the determination of intracellular cytokines IFN-γ+ (I), or TNF-α+ (J) from the spleen and lymph nodes at basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. Student t-test was performed for comparison between the two groups. Data are mean ± SEM (n = 5), ∗ p < 0.05.

    Article Snippet: Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection.

    Techniques: Isolation, Quantitative RT-PCR, Expressing, Western Blot, Knock-Out, Generated, Flow Cytometry, Comparison

    GPCR68 fl/fl CD4 Cre mice exhibit improved anti-tumor mmune responses. (A-C) Naïve CD4 + T cells were isolated from CD4 Cre or GPCR68 fl/fl CD4 Cre mice and activated using anti-CD3 and anti-CD28 using the culture media under physiologic neutral pH (7.4) or varying pH 6.0, 6.5, or 7.8. Flow cytometry plots showing the expression of IFN-γ and IL-2 in CD4 + T cells from CD4 Cre and GPCR68 fl/fl CD4 Cre mice. Each panel represents the frequency of IFN-γ + and IL-2 + cells. (B) Bar graph summarizing the percentage of IFN-γ + CD4 + T cells at each pH level for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (C) Bar graph showing the percentage of IL-2 + CD4 + T cells at each pH for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (D) Experimental timeline depicting tumor induction and treatment protocol in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (E) Tumor growth curves in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (F) Tumor weight in CD4 Cre versus GPCR68 fl/fl CD4 Cre mice at the time of harvesting on day 21. (G) Representative images of excised tumors at day 21. (H) Flow cytometric analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (I) Flow cytometric analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns = not significant.

    Journal: Bioactive Materials

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

    doi: 10.1016/j.bioactmat.2026.02.039

    Figure Lengend Snippet: GPCR68 fl/fl CD4 Cre mice exhibit improved anti-tumor mmune responses. (A-C) Naïve CD4 + T cells were isolated from CD4 Cre or GPCR68 fl/fl CD4 Cre mice and activated using anti-CD3 and anti-CD28 using the culture media under physiologic neutral pH (7.4) or varying pH 6.0, 6.5, or 7.8. Flow cytometry plots showing the expression of IFN-γ and IL-2 in CD4 + T cells from CD4 Cre and GPCR68 fl/fl CD4 Cre mice. Each panel represents the frequency of IFN-γ + and IL-2 + cells. (B) Bar graph summarizing the percentage of IFN-γ + CD4 + T cells at each pH level for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (C) Bar graph showing the percentage of IL-2 + CD4 + T cells at each pH for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (D) Experimental timeline depicting tumor induction and treatment protocol in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (E) Tumor growth curves in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (F) Tumor weight in CD4 Cre versus GPCR68 fl/fl CD4 Cre mice at the time of harvesting on day 21. (G) Representative images of excised tumors at day 21. (H) Flow cytometric analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (I) Flow cytometric analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns = not significant.

    Article Snippet: Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection.

    Techniques: Isolation, Flow Cytometry, Expressing, Comparison

    Anti-tumor effects of borate bioactive glass (BOLT) in B16 tumor. (A) Schematic illustration depicting the induction of B16 melanoma tumors, followed by treatment with BOLT at various time points, and tumor harvesting for subsequent analysis. (B) Tumor growth curves showing tumor volume in Control and BOLT-treated B16 melanoma tumors in mice. (C) Tumor weight at the time of harvesting in the BOLT-treated group compared to the Control. (D) Representative images of excised tumors from Control and BOLT-treated mice. (E) In vivo imaging of tumor-bearing mice in both the Control and BOLT-treated groups. (F) Flow cytometry analysis showing IFN-γ production in CD4 + and CD8 + T cells following BOLT treatment compared to Control. (G) Flow cytometry analysis demonstrated TNF-α production in CD4 + and CD8 + T cells in the BOLT-treated group, with a significant increase observed in CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data represent the mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01.

    Journal: Bioactive Materials

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

    doi: 10.1016/j.bioactmat.2026.02.039

    Figure Lengend Snippet: Anti-tumor effects of borate bioactive glass (BOLT) in B16 tumor. (A) Schematic illustration depicting the induction of B16 melanoma tumors, followed by treatment with BOLT at various time points, and tumor harvesting for subsequent analysis. (B) Tumor growth curves showing tumor volume in Control and BOLT-treated B16 melanoma tumors in mice. (C) Tumor weight at the time of harvesting in the BOLT-treated group compared to the Control. (D) Representative images of excised tumors from Control and BOLT-treated mice. (E) In vivo imaging of tumor-bearing mice in both the Control and BOLT-treated groups. (F) Flow cytometry analysis showing IFN-γ production in CD4 + and CD8 + T cells following BOLT treatment compared to Control. (G) Flow cytometry analysis demonstrated TNF-α production in CD4 + and CD8 + T cells in the BOLT-treated group, with a significant increase observed in CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data represent the mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01.

    Article Snippet: Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection.

    Techniques: Control, In Vivo Imaging, Flow Cytometry, Comparison

    Combinational treatment of BOLT and anti-CTLA-4 blockade enhances anti-tumor immune response in B16 melanoma. (A) C57BL/6 mice were subcutaneously injected with 1 × 10 5 B16 melanoma cells on day 0 to induce tumors. On day 7, mice were randomly divided into groups and treated with either BOLT alone (intratumoral injection administered on alternate days starting from day 7), anti-CTLA-4 (intraperitoneal injection administered on days 9, 11, 13, and 15), or a combination of both treatments. PBS was used as a vehicle Control, while IgG was used as anti-CTLA-4 Control. Tumor growth was monitored throughout the treatment period, and tumors were harvested for analysis on day 21. (B-C) Tumor growth curves and area under the curve (AUC) analysis for WT mice treated with BOLT, with or without anti-CTLA-4 antibody, following subcutaneous injection of B16 melanoma cells. Tumor growth was monitored, and analysis was conducted on day 21. (D) Representative images of excised tumors at day 21, showed reduced tumor size in combination-treated mice. (E, F) Flow cytometry analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (G, H) Flow cytometry analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

    Journal: Bioactive Materials

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

    doi: 10.1016/j.bioactmat.2026.02.039

    Figure Lengend Snippet: Combinational treatment of BOLT and anti-CTLA-4 blockade enhances anti-tumor immune response in B16 melanoma. (A) C57BL/6 mice were subcutaneously injected with 1 × 10 5 B16 melanoma cells on day 0 to induce tumors. On day 7, mice were randomly divided into groups and treated with either BOLT alone (intratumoral injection administered on alternate days starting from day 7), anti-CTLA-4 (intraperitoneal injection administered on days 9, 11, 13, and 15), or a combination of both treatments. PBS was used as a vehicle Control, while IgG was used as anti-CTLA-4 Control. Tumor growth was monitored throughout the treatment period, and tumors were harvested for analysis on day 21. (B-C) Tumor growth curves and area under the curve (AUC) analysis for WT mice treated with BOLT, with or without anti-CTLA-4 antibody, following subcutaneous injection of B16 melanoma cells. Tumor growth was monitored, and analysis was conducted on day 21. (D) Representative images of excised tumors at day 21, showed reduced tumor size in combination-treated mice. (E, F) Flow cytometry analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (G, H) Flow cytometry analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

    Article Snippet: Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection.

    Techniques: Injection, Control, Flow Cytometry