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naïve cd4 t cell isolation kit  (Miltenyi Biotec)


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    Miltenyi Biotec naïve cd4 t cell isolation kit
    Naïve Cd4 T Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 436 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+na%C3%AFve+cd4+t+cell+isolation+kit/Naive+CD4%2B+T+Cell+Isolation+Kit%2C+mouse/pmc13427790-70-17-25
    Average 97 stars, based on 436 article reviews
    naïve cd4 t cell isolation kit - by Bioz Stars, 2026-10
    97/100 stars

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    Related Articles

    Red Blood Cell Lysis:

    Article Title: Elephantopus scaber suppresses ConA-induced T cell activation by inhibiting NF-κB and Ca 2+ /NFAT signaling.
    Article Snippet: Ethnopharmacological relevance: Elephantopus scaber L. (E. scaber) is a heat-clearing and detoxifying traditional Chinese medicine.. Clinically, it is used for treating some immuno-inflammatory diseases associated with excessive T cell activation.. However, its action and mechanism on T cells are still ambiguous.

    Cell Isolation:

    Article Title: Elephantopus scaber suppresses ConA-induced T cell activation by inhibiting NF-κB and Ca 2+ /NFAT signaling.
    Article Snippet: Ethnopharmacological relevance: Elephantopus scaber L. (E. scaber) is a heat-clearing and detoxifying traditional Chinese medicine.. Clinically, it is used for treating some immuno-inflammatory diseases associated with excessive T cell activation.. However, its action and mechanism on T cells are still ambiguous.

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    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. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Article Title: Molecular Engineering of the Helminth <scp>TGF</scp> ‐β Mimetics, <scp>TGM1</scp> and <scp>TGM4,</scp> Reveals a Novel Antagonist of <scp>TGF</scp> ‐β Signaling in Fibroblasts
    Article Snippet: Single cell suspensions of naïve BALB/c or Foxp3- green fluorescent protein (GFP) BALB/c transgenic [24] mouse splenocytes were prepared and incubated for 2 min in red blood cell lysis buffer (Sigma) before washing and resuspending in RPMI1640 containing HEPES (Gibco) 2 mM L- glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), 10% heat- inactivated FBS (Gibco), and 50 nM 2- mercaptoethanol (Gibco). .. Naïve CD4+ T cells were then isolated by magnetic sorting using the mouse naïve CD4+ T cell isolation kit on the AutoMACS system (Miltenyi, Germany) as per the manufacturer's instructions. .. For Foxp3 induction, flat- bottomed 96- well plates (Corning, USA) pre- coated with 10 μg/mL of anti- CD3 (eBioscience), were populated with 2 × 105 cells per well in media supplemented with 400 U/mL interleukin (IL)- 2 (Miltenyi).

    Article Title: Actomyosin rings constrain CD40 mobility to organize the dendritic cell immunological synapse.
    Article Snippet: The resulting cell suspension was centrifuged at 300 × g for 10 minutes and then resuspended in staining buffer (PBS containing 2% iFCS and 2 mM EDTA (PanReac Aplichem, A3145-0500). .. T cells were further purified using the Mouse Naïve CD4+ T Cell Isolation Kit (Miltenyi Biotec, 130-104-453), following the manufacturer’s protocol. .. After isolation, the cells were cultured in R10 medium (RPMI 1640 with 2 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, 10% heat-inactivated fetal calf serum (iFCS), and 50 μM 2- mercaptoethanol) supplemented with 0.4 U IL-7 (Peprotech, 217-17) and incubated at 37°C with 5% CO2.

    Article Title: UBE2O primes hepatocytes to restore immune tolerance in autoimmune hepatitis via inhibiting YBX1/IL-6 axis.
    Article Snippet: .. The next steps were according to 625 the manufacturer’s instructions by a mouse Naïve CD4+T Cell Isolation Kit (cat#130-626 104-453, Miltenyi Biotec). ..

    Article Title: Ubiquitin-Conjugating Enzyme E2O Primes Hepatocytes to Restore Immune Tolerance in Autoimmune Hepatitis via Inhibiting Y-Box Binding Protein 1/Interleukin-6 Axis
    Article Snippet: .. The next steps were according to the manufacturer’s instructions by a mouse Naïve CD4 + T Cell Isolation Kit (cat#130-104-453, Miltenyi Biotec). ..

    Article Title: GPSM1 restricts CD73 + CD103 + T reg cells in adipose tissue, critical for promoting obesity-related metabolic deterioration
    Article Snippet: .. Naïve CD4 + T cells were isolated from splenic and peripheral lymph node cells of mice using the mouse naïve CD4 + T Cell Isolation Kit (Cat. No. 130-104-453, Miltenyi Biotec). .. The isolated naïve CD4 + T cells were cultured in 12-well or 24-well plates pre-incubated overnight with anti-mouse CD3e (2 μg/mL, Cat. No. 553057, BD Biosciences).

    Article Title: Molecular Engineering of the Helminth TGF ‐β Mimetics, TGM1 and TGM4, Reveals a Novel Antagonist of TGF ‐β Signaling in Fibroblasts
    Article Snippet: Single cell suspensions of naïve BALB/c or Foxp3‐green fluorescent protein (GFP) BALB/c transgenic [ ] mouse splenocytes were prepared and incubated for 2 min in red blood cell lysis buffer (Sigma) before washing and resuspending in RPMI1640 containing HEPES (Gibco) 2 mM L‐glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), 10% heat‐inactivated FBS (Gibco), and 50 nM 2‐mercaptoethanol (Gibco). .. Naïve CD4 + T cells were then isolated by magnetic sorting using the mouse naïve CD4 + T cell isolation kit on the AutoMACS system (Miltenyi, Germany) as per the manufacturer's instructions. .. For Foxp3 induction, flat‐bottomed 96‐well plates (Corning, USA) pre‐coated with 10 μg/mL of anti‐CD3 (eBioscience), were populated with 2 × 10 5 cells per well in media supplemented with 400 U/mL interleukin (IL)‐2 (Miltenyi).

    Purification:

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    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. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Article Title: Actomyosin rings constrain CD40 mobility to organize the dendritic cell immunological synapse.
    Article Snippet: The resulting cell suspension was centrifuged at 300 × g for 10 minutes and then resuspended in staining buffer (PBS containing 2% iFCS and 2 mM EDTA (PanReac Aplichem, A3145-0500). .. T cells were further purified using the Mouse Naïve CD4+ T Cell Isolation Kit (Miltenyi Biotec, 130-104-453), following the manufacturer’s protocol. .. After isolation, the cells were cultured in R10 medium (RPMI 1640 with 2 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, 10% heat-inactivated fetal calf serum (iFCS), and 50 μM 2- mercaptoethanol) supplemented with 0.4 U IL-7 (Peprotech, 217-17) and incubated at 37°C with 5% CO2.

    Selection:

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    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. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Isolation:

    Article Title: Molecular Engineering of the Helminth <scp>TGF</scp> ‐β Mimetics, <scp>TGM1</scp> and <scp>TGM4,</scp> Reveals a Novel Antagonist of <scp>TGF</scp> ‐β Signaling in Fibroblasts
    Article Snippet: Single cell suspensions of naïve BALB/c or Foxp3- green fluorescent protein (GFP) BALB/c transgenic [24] mouse splenocytes were prepared and incubated for 2 min in red blood cell lysis buffer (Sigma) before washing and resuspending in RPMI1640 containing HEPES (Gibco) 2 mM L- glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), 10% heat- inactivated FBS (Gibco), and 50 nM 2- mercaptoethanol (Gibco). .. Naïve CD4+ T cells were then isolated by magnetic sorting using the mouse naïve CD4+ T cell isolation kit on the AutoMACS system (Miltenyi, Germany) as per the manufacturer's instructions. .. For Foxp3 induction, flat- bottomed 96- well plates (Corning, USA) pre- coated with 10 μg/mL of anti- CD3 (eBioscience), were populated with 2 × 105 cells per well in media supplemented with 400 U/mL interleukin (IL)- 2 (Miltenyi).

    Article Title: GPSM1 restricts CD73 + CD103 + T reg cells in adipose tissue, critical for promoting obesity-related metabolic deterioration
    Article Snippet: .. Naïve CD4 + T cells were isolated from splenic and peripheral lymph node cells of mice using the mouse naïve CD4 + T Cell Isolation Kit (Cat. No. 130-104-453, Miltenyi Biotec). .. The isolated naïve CD4 + T cells were cultured in 12-well or 24-well plates pre-incubated overnight with anti-mouse CD3e (2 μg/mL, Cat. No. 553057, BD Biosciences).

    Article Title: Molecular Engineering of the Helminth TGF ‐β Mimetics, TGM1 and TGM4, Reveals a Novel Antagonist of TGF ‐β Signaling in Fibroblasts
    Article Snippet: Single cell suspensions of naïve BALB/c or Foxp3‐green fluorescent protein (GFP) BALB/c transgenic [ ] mouse splenocytes were prepared and incubated for 2 min in red blood cell lysis buffer (Sigma) before washing and resuspending in RPMI1640 containing HEPES (Gibco) 2 mM L‐glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), 10% heat‐inactivated FBS (Gibco), and 50 nM 2‐mercaptoethanol (Gibco). .. Naïve CD4 + T cells were then isolated by magnetic sorting using the mouse naïve CD4 + T cell isolation kit on the AutoMACS system (Miltenyi, Germany) as per the manufacturer's instructions. .. For Foxp3 induction, flat‐bottomed 96‐well plates (Corning, USA) pre‐coated with 10 μg/mL of anti‐CD3 (eBioscience), were populated with 2 × 10 5 cells per well in media supplemented with 400 U/mL interleukin (IL)‐2 (Miltenyi).



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    naïve cd4 t cell isolation kit  (Miltenyi Biotec)
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    Miltenyi Biotec naïve cd4 t cell isolation kit
    Naïve Cd4 T Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse naïve cd4 t cell isolation kit  (Miltenyi Biotec)
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    GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl <t>CD4</t> 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.
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    Expected results of this protocol <t>Naïve</t> CD4 + T cells cultured for d5 in supernatant of untreated BMDCs (SN NT) or in supernatant of BPI-treated BMDCs (SN BPI). (A) Representative dot blot of flow cytometric analysis of CD62L and CD44 cell surface presentation. (B) IL-22 secretion measured by Luminex technology, n = 4. Data are shown as means ± SEM. Statistical testing was performed using Student`s ratio paired t test.
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    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

    Physicochemical properties of BOLT, and BOLT reduces the growth of tumor cells. (A) Schematic of surface double-layer formation and ion release. (B) Negative zeta potential (−1.365 mV) and high conductivity (1.334 mS/cm), confirming colloidal stability and ion release. (C) Uniform particle size (∼1478 nm) across batches. (D) Interfacial pH buffering in PBS. (E) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with 6.0 pH and treated with various doses of BOLT. RT-qPCR was performed to determine the expression of Gpcr68 at various BOLT doses in activated T cells at acidic pH. (F) Anti-CD3 and anti-CD28 activated CD4 + T cells were treated with different doses of BOLT to determine the protein expression of GPCR68 using Western blot. (G-J) CCK8 assay was performed to analyze the effect of various pH on B16, MC38, 143B, and MG63 cell proliferation. (K-L) Effect of various doses of BOLT on the B16 and K7M2 cell growth to determine the IC-50 of BOLT. Error bars represent mean ± SEM. ∗∗ p < 0.01 and ∗ 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: Physicochemical properties of BOLT, and BOLT reduces the growth of tumor cells. (A) Schematic of surface double-layer formation and ion release. (B) Negative zeta potential (−1.365 mV) and high conductivity (1.334 mS/cm), confirming colloidal stability and ion release. (C) Uniform particle size (∼1478 nm) across batches. (D) Interfacial pH buffering in PBS. (E) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with 6.0 pH and treated with various doses of BOLT. RT-qPCR was performed to determine the expression of Gpcr68 at various BOLT doses in activated T cells at acidic pH. (F) Anti-CD3 and anti-CD28 activated CD4 + T cells were treated with different doses of BOLT to determine the protein expression of GPCR68 using Western blot. (G-J) CCK8 assay was performed to analyze the effect of various pH on B16, MC38, 143B, and MG63 cell proliferation. (K-L) Effect of various doses of BOLT on the B16 and K7M2 cell growth to determine the IC-50 of BOLT. Error bars represent mean ± SEM. ∗∗ p < 0.01 and ∗ 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: Zeta Potential Analyzer, Isolation, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay

    BOLT activates T cell PI3K-AKT-mTOR pathway to enhance T cell anti-tumor effect. (A) Flow cytometry plots compare IFN-γ and IL-2 expression at pH 7.8 and 6.0 along with various doses of BOLT in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (B, C) Bar graphs show IFN-γ and IL-2 expression in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (D) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 antibodies and incubated for 3 days with cell culture media of different pH levels. Western blot was performed to determine the phosphorylation of Akt and S6 under acidic conditions (pH 6.5) and alkaline pH (7.8). (E) Activated CD4 + T cells were treated with 0, 0.25, and 0.5 mg/mL doses of BOLT following CD4 + T cells activation at pH 7.8. Western blot analysis showing the phosphorylation of Akt and S6 were observed. (F) CD4 + T cells were activated and treated with BOLT at acidic pH. Western blot analysis was performed to determine the phosphorylation of Akt and S6. Two-way ANOVA was used for multiple comparisons. Experiments were conducted in triplicate. Data are mean ± SEM, ∗ 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: BOLT activates T cell PI3K-AKT-mTOR pathway to enhance T cell anti-tumor effect. (A) Flow cytometry plots compare IFN-γ and IL-2 expression at pH 7.8 and 6.0 along with various doses of BOLT in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (B, C) Bar graphs show IFN-γ and IL-2 expression in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (D) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 antibodies and incubated for 3 days with cell culture media of different pH levels. Western blot was performed to determine the phosphorylation of Akt and S6 under acidic conditions (pH 6.5) and alkaline pH (7.8). (E) Activated CD4 + T cells were treated with 0, 0.25, and 0.5 mg/mL doses of BOLT following CD4 + T cells activation at pH 7.8. Western blot analysis showing the phosphorylation of Akt and S6 were observed. (F) CD4 + T cells were activated and treated with BOLT at acidic pH. Western blot analysis was performed to determine the phosphorylation of Akt and S6. Two-way ANOVA was used for multiple comparisons. Experiments were conducted in triplicate. Data are mean ± SEM, ∗ 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: Flow Cytometry, Expressing, Incubation, Cell Culture, Western Blot, Phospho-proteomics, Activation Assay

    Expected results of this protocol Naïve CD4 + T cells cultured for d5 in supernatant of untreated BMDCs (SN NT) or in supernatant of BPI-treated BMDCs (SN BPI). (A) Representative dot blot of flow cytometric analysis of CD62L and CD44 cell surface presentation. (B) IL-22 secretion measured by Luminex technology, n = 4. Data are shown as means ± SEM. Statistical testing was performed using Student`s ratio paired t test.

    Journal: STAR Protocols

    Article Title: Protocol for potent activation of T cells using BPI-stimulated murine bone marrow-derived cells

    doi: 10.1016/j.xpro.2026.104519

    Figure Lengend Snippet: Expected results of this protocol Naïve CD4 + T cells cultured for d5 in supernatant of untreated BMDCs (SN NT) or in supernatant of BPI-treated BMDCs (SN BPI). (A) Representative dot blot of flow cytometric analysis of CD62L and CD44 cell surface presentation. (B) IL-22 secretion measured by Luminex technology, n = 4. Data are shown as means ± SEM. Statistical testing was performed using Student`s ratio paired t test.

    Article Snippet: Note: If no FACS device is available, the sorting of naïve T cells can be performed using a naïve MACS Sort Kit from Miltenyi Biotec (130-104-453 ).

    Techniques: Cell Culture, Dot Blot, Luminex