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untouched splenic cd8 t splenocytes  (Miltenyi Biotec)


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    Miltenyi Biotec untouched splenic cd8 t splenocytes
    Untouched Splenic Cd8 T Splenocytes, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 630 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd8a+t+cell+isolation+kit/CD8a%2B+T+Cell+Isolation+Kit%2C+mouse/10__1158_slash_1535___7163__mct___25___1488-143-7-26
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    untouched splenic cd8 t splenocytes - by Bioz Stars, 2026-10
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    Related Articles

    Isolation:

    Article Title: Multiplexed cytokine and antigen mRNA administration generates durable anti-tumor immunity against pancreatic cancer
    Article Snippet: After 4 h of incubation at 37 °C in a humidified incubator with 5% CO2, migrated cells in the lower chamber were fixed with 4% paraformaldehyde (PFA, ThermoFisher Scientific, J61899.AK), stained with DAPI (Millipore Sigma, D9542), and quantified using a Celigo imaging cytometer (Nexcelom). .. Primary NK and CD8+ T cells were isolated from spleens of 8- to 12-week-old naïve female C57BL/6 mice using the NK Cell Isolation Kit II (Miltenyi Biotec; Cat. No 130-115-818) and CD8a+ T Cell Isolation Kit (Miltenyi Biotec; Cat. No 130-104-075), respectively, according to the manufacturer’s instructions. .. NK or CD8+ T cells were then co-cultured at a 5:1 effector-to-target ratio with KPC1 PDAC tumor cells transfected 24 h earlier with different single or combinatorial mRNA cocktails in RPMI media (Corning, 10-040-CV) supplemented with 10% FBS, PMA (20 ng/ml, STEMCELL Technologies, 74042), Ionomycin (1 μg/ml, STEMCELL technologies, 73722), and monensin (2 μM, Biolegend, 420701) in a humidified incubator at 37 °C with 5% CO 2 .

    Article Title: A tumor-intrinsic WNT-inhibitory NOTUM program drives immune resistance in microsatellite stable colorectal cancer
    Article Snippet: A portion of the resuspended pellet was mixed with trypan blue (Gibco) and counted on a hemocytometer to determine T cell quantity and viability. .. Following this, CD8a+ cells were selectively isolated using the MidiMACS Separator and Starting Kit and the CD8a+ T cell Isolation Kit (Miltenyi Biotec, #130-104-075 and #130-090-329 respectively), according to the manufacturer’s instructions. .. CD8 + T cells were plated on 6 well plates (Falcon) coated with anti-CD3 and anti-CD28 (BD Bioscience).

    Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model
    Article Snippet: All images were acquired using an LSM5 PASCAL microscope (Carl Zeiss, Jena, Germany), and total staining intensity was quantified using ImageJ software (NIH) ( ). .. CD8 + T cells were isolated from the spleens of 6-week-old C57BL/6 mice using a CD8a + T Cell Isolation Kit (Miltenyi Biotec, Cat. No. 130-104-075) according to the manufacturer’s instructions. .. For T cell activation, isolated CD8 + T cells were cultured in plates coated with anti-CD3 antibody (5 μg/mL; BD biosciences, Cat. No. 553057) and supplemented with soluble anti-CD28 antibody (1 μg/mL) for 24 h. For the co-culture experiment, M2-polarized RAW264.7 macrophages were co-cultured with CD8 + T cells and treated with TAMpep-IP (0.5 μM).

    Article Title: Pancreatic cancer induces B cell lineage plasticity via Pax5 inhibition to sustain immunosuppression.
    Article Snippet: .. On day five, CD3+CD8a+ T cells were isolated from the spleens of wt C57BL/6 J mice using a CD8a+ T cell isolation kit (Miltenyi Biotec, Cat. No.130-104-075) according to the manufacturer’s instructions. .. Depending on the experimental condition, the T cells were stimulated using mouse T-activator CD3/CD28 Dynabeads (Thermo Fisher Scientific, Cat. No.11456D) according to the manufacturer’s instructions.

    Article Title: Targeting CCR1 remodels the tumor microenvironment and relieves immune suppression in pancreatic cancer
    Article Snippet: Flow cytometry data acquisition was performed on LSRFortessa Cell Analyzer (BD Biosciences, RRID:SCR_018655) and analyzed using FlowJo software v10 (FlowJo, RRID:SCR_008520). .. CD8+ T cell migration assay CD8+ T cells were isolated from the spleens and lymph nodes of WT mice (C57BL/6J, Jackson Laboratory, strain #000664, RRID:IMSR_JAX:000664) by CD8a+ T Cell Isolation Kit (Miltenyi Biotec, catalog #130-104-075). .. Isolated CD8 T cells were resuspended in 10% serum RPMI 1640 media and 8 × 105 cells in 800 μl media were plated to the top transwell (3 μm pore size) of a 6- well plate (Corning, catalog #3452).

    Cell Isolation:

    Article Title: Multiplexed cytokine and antigen mRNA administration generates durable anti-tumor immunity against pancreatic cancer
    Article Snippet: After 4 h of incubation at 37 °C in a humidified incubator with 5% CO2, migrated cells in the lower chamber were fixed with 4% paraformaldehyde (PFA, ThermoFisher Scientific, J61899.AK), stained with DAPI (Millipore Sigma, D9542), and quantified using a Celigo imaging cytometer (Nexcelom). .. Primary NK and CD8+ T cells were isolated from spleens of 8- to 12-week-old naïve female C57BL/6 mice using the NK Cell Isolation Kit II (Miltenyi Biotec; Cat. No 130-115-818) and CD8a+ T Cell Isolation Kit (Miltenyi Biotec; Cat. No 130-104-075), respectively, according to the manufacturer’s instructions. .. NK or CD8+ T cells were then co-cultured at a 5:1 effector-to-target ratio with KPC1 PDAC tumor cells transfected 24 h earlier with different single or combinatorial mRNA cocktails in RPMI media (Corning, 10-040-CV) supplemented with 10% FBS, PMA (20 ng/ml, STEMCELL Technologies, 74042), Ionomycin (1 μg/ml, STEMCELL technologies, 73722), and monensin (2 μM, Biolegend, 420701) in a humidified incubator at 37 °C with 5% CO 2 .

    Article Title: A tumor-intrinsic WNT-inhibitory NOTUM program drives immune resistance in microsatellite stable colorectal cancer
    Article Snippet: A portion of the resuspended pellet was mixed with trypan blue (Gibco) and counted on a hemocytometer to determine T cell quantity and viability. .. Following this, CD8a+ cells were selectively isolated using the MidiMACS Separator and Starting Kit and the CD8a+ T cell Isolation Kit (Miltenyi Biotec, #130-104-075 and #130-090-329 respectively), according to the manufacturer’s instructions. .. CD8 + T cells were plated on 6 well plates (Falcon) coated with anti-CD3 and anti-CD28 (BD Bioscience).

    Article Title: Newly isolated human-derived Lactobacillus fermentum strain stimulates IFN-γ-secreting CD8 + T cells for enhanced anti-cancer immunity
    Article Snippet: CD11c MicroBeads UltraPure, mouse , Miltenyi Biotec , 130-125-835. .. CD8a+ T cell Isolation Kit, mouse , Miltenyi Biotec , 130-104-075. .. PiPerTM Phosphate Assay Kit , Invitrogen , P22061.

    Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model
    Article Snippet: All images were acquired using an LSM5 PASCAL microscope (Carl Zeiss, Jena, Germany), and total staining intensity was quantified using ImageJ software (NIH) ( ). .. CD8 + T cells were isolated from the spleens of 6-week-old C57BL/6 mice using a CD8a + T Cell Isolation Kit (Miltenyi Biotec, Cat. No. 130-104-075) according to the manufacturer’s instructions. .. For T cell activation, isolated CD8 + T cells were cultured in plates coated with anti-CD3 antibody (5 μg/mL; BD biosciences, Cat. No. 553057) and supplemented with soluble anti-CD28 antibody (1 μg/mL) for 24 h. For the co-culture experiment, M2-polarized RAW264.7 macrophages were co-cultured with CD8 + T cells and treated with TAMpep-IP (0.5 μM).

    Article Title: Pancreatic cancer induces B cell lineage plasticity via Pax5 inhibition to sustain immunosuppression.
    Article Snippet: .. On day five, CD3+CD8a+ T cells were isolated from the spleens of wt C57BL/6 J mice using a CD8a+ T cell isolation kit (Miltenyi Biotec, Cat. No.130-104-075) according to the manufacturer’s instructions. .. Depending on the experimental condition, the T cells were stimulated using mouse T-activator CD3/CD28 Dynabeads (Thermo Fisher Scientific, Cat. No.11456D) according to the manufacturer’s instructions.

    Article Title: Targeting CCR1 remodels the tumor microenvironment and relieves immune suppression in pancreatic cancer
    Article Snippet: Flow cytometry data acquisition was performed on LSRFortessa Cell Analyzer (BD Biosciences, RRID:SCR_018655) and analyzed using FlowJo software v10 (FlowJo, RRID:SCR_008520). .. CD8+ T cell migration assay CD8+ T cells were isolated from the spleens and lymph nodes of WT mice (C57BL/6J, Jackson Laboratory, strain #000664, RRID:IMSR_JAX:000664) by CD8a+ T Cell Isolation Kit (Miltenyi Biotec, catalog #130-104-075). .. Isolated CD8 T cells were resuspended in 10% serum RPMI 1640 media and 8 × 105 cells in 800 μl media were plated to the top transwell (3 μm pore size) of a 6- well plate (Corning, catalog #3452).

    Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells
    Article Snippet: Lung Dissociation kit , Miltenyi Biotech , Cat# 130-095-927; RRID: SCR_020286. .. CD8a+ T cell isolation kit , Miltenyi Biotec , Cat#130-104-075. .. Foxp3/Transcription Factor Staining Buffer Set kit , eBioscience , Cat#00-5523-00.

    Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells
    Article Snippet: After 7 days, NP68 peptide (20 nM) and CpG ODN 1826 (2 mg/mL, InvivoGen, tlrl-1826) were added to the BMDC and cultured overnight before being washed and used for CD8 + T cells activation. .. Total CD8 + T cells were enriched from splenocytes using an autoMACS Pro Separator (Miltenyi Biotec, RRID: SCR_018596 ) with the CD8a+ T cell isolation kit (Miltenyi Biotec, 130-104-075) according to the manufacturer’s instructions. ..

    Cell Migration Assay:

    Article Title: Targeting CCR1 remodels the tumor microenvironment and relieves immune suppression in pancreatic cancer
    Article Snippet: Flow cytometry data acquisition was performed on LSRFortessa Cell Analyzer (BD Biosciences, RRID:SCR_018655) and analyzed using FlowJo software v10 (FlowJo, RRID:SCR_008520). .. CD8+ T cell migration assay CD8+ T cells were isolated from the spleens and lymph nodes of WT mice (C57BL/6J, Jackson Laboratory, strain #000664, RRID:IMSR_JAX:000664) by CD8a+ T Cell Isolation Kit (Miltenyi Biotec, catalog #130-104-075). .. Isolated CD8 T cells were resuspended in 10% serum RPMI 1640 media and 8 × 105 cells in 800 μl media were plated to the top transwell (3 μm pore size) of a 6- well plate (Corning, catalog #3452).



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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