Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Dynamic changes in serum E2 levels and intrahepatic CD4 + CD25 + Foxp3 + Tregs in HBV-Tg mice during pregnancy and postpartum. a. One representative dot plot from each time point is shown. b. The proportion of CD4 + CD25 + Foxp3 + Tregs among CD4 + cells was compared. c. Dynamic changes in the serum E2 concentration were observed. The results were analyzed via one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Concentration Assay

Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Exogenous E2 injection in ovariectomized HBV-Tg mice promoted the proliferation of intrahepatic CD4 + CD25 + Foxp3 + Tregs. (a) An illustration of the study design. (b) Representative diagram of the proportion of intrahepatic CD4 + CD25 + Foxp3 + Tregs among the three groups. (c) Comparison of the proportion of intrahepatic CD4 + CD25 + Foxp3 + Tregs among the three groups. The results were compared and analyzed using via one-way ANOVA. ** p < 0.01

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Injection, Comparison

Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Estrogen plays a protective role against postpartum Con A-induced hepatitis, with increased intrahepatic CD4 + CD25 + Foxp3 + Tregs and inhibitory cytokine IL-10 levels. (a) HBV-Tg mice were i.p. injected with E2 to maintain the serum estrogen level as high as that observed in late pregnancy or sesame oil as control. On postpartum day 7, Con A was injected i.v. and peripheral blood and liver tissues were collected for further analysis 24 h after Con A administration. (b) Serum ALT levels in the Control, Con A, Oil + Con A and E2 + Con A groups. (c) The percentages of intrahepatic CD4 + CD25 + Foxp3 + Tregs were determined for different groups. (d) The inhibitory cytokine IL-10 in serum of different groups were measured via ELISA, and its levels in the liver was measured via qRT-PCR (mRNA level) and ELISA (protein level). (e) CD107a expression in CD8 + T cells within hepatic MNCs were analyzed. (f) Proliferation of CD8 + T cells within hepatic MNCs were assessed. (g) The expression levels of the activation marker CD69 on both NK cells and NKT cells were determined. (h) The levels of the cytotoxic cytokine IFN-γ in serum were measured via ELISA, and its levels in the liver were measured via qRT-PCR (mRNA level) and ELISA (protein level). (i) The levels of the cytotoxic cytokine TNF-α in serum were measured via ELISA, and its levels in the liver were measured via qRT-PCR (mRNA level) and ELISA (protein level). The results were compared among the different groups via two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Injection, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Activation Assay, Marker

Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Intrahepatic CD4 + CD25 + Foxp3 + Tregs play a pivotal role in the protective effect of E2 against postpartum Con A-induced hepatitis. Depletion of hepatic CD25 + cells aggravated liver damage in estrogen-injected HBV-Tg mice. a. The mice were injected E2 (100 ng/mouse) every other day starting from delivery day, then Con A (2 µg/g) injected on postpartum day 7, and 6 h after Con A treatment, a dose of 100 µg of rat IgG1 kappa isotype control antibody or 100 µg/mouse of anti-CD25 mAbs was injected i.p. 24 h post-Con A administration, peripheral blood and liver tissues were collected for analysis. b . ALT levels in serum were measured in mice administered with anti-CD25 Abs, compared to those given isotype control Abs. (c) Comparison of intrahepatic CD4 + CD25 + Foxp3 + Tregs between mice injected with anti-CD25 Abs and those injected with isotype control Abs. (d) The inhibitory cytokine IL-10 in serum of different groups were measured via ELISA, and its levels in the liver was measured via qRT-PCR (mRNA level) and ELISA (protein level). (e) CD107a expression in CD8 + T cells within hepatic MNCs were analyzed. (f) Proliferation of CD8 + T cells within hepatic MNCs were assessed. (g) Expression of the activation marker CD69 on NK cells and NKT cells were determined. (h) The levels of the cytotoxic cytokine IFN-γ in serum were measured via ELISA, and its levels in the liver was measured via qRT-PCR (mRNA level) and ELISA (protein level). (i) The levels of the cytotoxic cytokine TNF-α in serum were measured via ELISA, and its levels in the liver was measured via qRT-PCR (mRNA level) and ELISA (protein level). The results were analyzed by two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Injection, Control, Comparison, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Activation Assay, Marker

Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Administering E2 resulted in elevated p-Akt levels of purified hepatic CD25 + cells, and the PI3K/Akt signaling pathway was blocked via i.p. injection of the PI3K inhibitor LY294002 at the same time as E2 treatment in vivo. (a) An illustration of the study design. (b) The expressions of p-Akt and Akt in hepatic CD25 + cells were evaluated across the three groups (Control group, Oil + Con A group, and E2 + Con A group). (c) The expressions of p-Akt and Akt in hepatic CD25 + cells were evaluated across the four groups (E2 + Con A group, LY294002 group, E2 + Con A + LY294002 group and E2 + Con A + Solvent group)

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Purification, Injection, In Vivo, Control, Solvent

Journal: Virology Journal

Article Title: Estrogen protects against postpartum Concanavalin A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice

doi: 10.1186/s12985-025-02879-4

Figure Lengend Snippet: Estrogen protects against postpartum Con A-induced hepatitis by promoting intrahepatic CD4⁺CD25⁺ Treg expansion through activation of the PI3K/Akt signaling pathway in HBV-Tg mice. (a) The PI3K inhibitor LY294002 or solvent DMSO was i.p. injected at the same time as the E2 treatment. The peripheral blood and liver tissues were collected for further analysis. (b) Serum ALT concentrations were determined in LY294002-treated mice compared to mice that received the solvent control. (c) The percentage of intrahepatic CD4 + CD25 + Foxp3 + Tregs in the livers of mice treated with LY294002 compared with those in the livers of control DMSO-treated mice. (d) The inhibitory cytokine IL-10 in serum of different groups were measured via ELISA, and its levels in the liver were measured via qRT-PCR (mRNA level) and ELISA (protein level). e. CD107a expression in CD8 + T cells in hepatic MNCs were analyzed. f. Proliferation of CD8 + T cells in hepatic MNCs were assessed. g. Expression of the activation marker CD69 on NK cells and NKT cells were determined. h. The cytotoxic cytokine IFN-γ in serum was measured via ELISA, and its level in the liver were measured via qRT-PCR (mRNA level) and ELISA (protein level). i. The levels of the cytotoxic cytokine TNF-α in serum were measured via ELISA, and its levels in the liver were measured via qRT-PCR (mRNA level) and ELISA (protein level). The results were analyzed via two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: We utilized a CD4 + CD25 + Regulatory T-Cell Isolation Kit (130-091-041; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) to isolate CD25 + hepatic MNCs.

Techniques: Activation Assay, Solvent, Injection, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Marker

Journal: Aging Cell

Article Title: Role of Succinate Dehydrogenase in Age‐Related Th17 Inflammation

doi: 10.1111/acel.70451

Figure Lengend Snippet: Succinate dehydrogenase protein expression increases with age in CD4 + T cells. SDH expression in young (Y) and older (O) adults assessed via immunoblotting (a) and confocal microscopy (b). SDHA protein expression in CD4 + T cells treated with SDH inhibitor 3‐nitropropionic acid (3NP) and cell‐permeable diethyl succinate (DES); immunoblotting (c) and microscopy (d). SDHB protein expression after the different treatments (e). N = 3–4 (a–e). N = 3–4 indicates cells were obtained from either three or four individuals for each condition. Microscopy data are represented as cells in the field of view. At least 5–7 fields per slide were imaged at 63× magnification with oil immersion, on a Zeiss LSM 800 confocal microscope. In fields where numerous cells were observed, the mean fluorescence intensity of 3–4 cell groups was plotted as a single dot. Images were processed as described in the methods, and brightness was adjusted to improve clarity. Mann–Whitney Test or Kruskal–Wallis test with Dunn's post hoc test, * p < 0.05 vs. Y. # p < 0.05 vs. O or O + 3NP.

Article Snippet: Human CD4 + isolation kit , Miltenyi , Cat no. 130‐096‐533.

Techniques: Expressing, Western Blot, Confocal Microscopy, Microscopy, Fluorescence, MANN-WHITNEY

Journal: Aging Cell

Article Title: Role of Succinate Dehydrogenase in Age‐Related Th17 Inflammation

doi: 10.1111/acel.70451

Figure Lengend Snippet: Age‐induced dysregulation of TCA cycle metabolites fuel Th17 cytokine production. Waterfall plots showing ScRNA seq analysis of TCA cycle enzymes in CD4 + T cells from young (Y) and older (O) adults (a) ScRNA seq analysis of TCA cycle enzymes in Th17 subset of T cells (b). Cellular amounts of succinate (c) fumarate:succinate ratio (d) HIF1α protein in T cells from older adults (e) and HIF1α protein in T cells from younger adults after FH inhibition (f) N = 3, (a, b) N = 3–4, (c, d) N = 5–7, (e) N = 3, (f) adults in each group. One‐way ANOVA with Bonferroni test or Wilcoxon matched‐pair signed rank test. * p < 0.05 vs. O or Y.

Article Snippet: Human CD4 + isolation kit , Miltenyi , Cat no. 130‐096‐533.

Techniques: Inhibition

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: C57BL/6 mice were treated with CPT-11, and immune responses were determined by flow cytometry (FCM). a Total number of immune cells in spleen and lymph nodes (LNs) of mice treated with PBS (control) or CPT-11 ( n = 4 mice per group). b , c Bar graphs show the frequency of Ki67 + CD4 + and Ki67 + CD8 + T cells in the indicated groups. d Representative fluorescence-activated cell sorting (FACS) plots of indicated groups. e – j Bar graphs showing frequencies of IFN-γ + CD4 + T (Th1) cells, T-bet + CD4 + T (Th1) cells, IFN-γ + CD8 + T cells, IL-17 + CD4 + T (Th17) cells, IL-4 + CD4 + T (Th2) cells, and FoxP3 + CD4 + Treg cells in indicated groups. Data are representative of two independent experiments. Summary data are presented as mean ± s.d. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; by unpaired two-tailed Student’s t-tests. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Flow Cytometry, Control, Fluorescence, FACS, Two Tailed Test

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: C57BL/6 mice were challenged with CFA (subcutaneous injection) and treated with CPT-11 or PBS, and immune responses in spleen and LNs were determined using FCM. a Total number of immune cells in the spleen and LNs of mice treated with PBS (control) or CPT-11. ( n = 4 mice per group). b Representative FACS plots of indicated groups. c – g Bar graphs showing frequencies of Ki67 + CD4 + and Ki67 + CD8 + T cells, IFN-γ + CD4 + Th1 cells, IL-17 + CD4 + Th17 cells, and IFN-γ + CD8 + cells from indicated mice. Data are representative of two independent experiments. Summary data are presented as mean ± s.d. ** p < 0.01, *** p < 0.001, **** p < 0.0001; by unpaired two-tailed Student’s t-tests. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Injection, Control, Two Tailed Test

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: CD4 + CD25 − CD62L high (naive) T cells isolated from spleen and LNs of C57BL/6 mice were cultured with anti-CD3 and anti-CD28, with or without CPT-11 for 1-3 d. Cell proliferation, cell apoptosis, and cell differentiation were determined using FCM ( n = 3). a , b Representative FACS plots ( a ) and bar graph ( b ) showing non-proliferative T cell frequencies in T cells cultured for 3 d. c , d Representative FACS plots ( c ) and bar graph ( d ) showing apoptotic T cell frequencies in T cells cultured for 24 h. e , f Representative FACS plots ( e ) and bar graph ( f ) showing the frequency of Th1 cells in T cells cultured for 3 d in the presence of IL-12. g , h Representative FACS plots ( g ) and bar graph ( h ) showing frequencies of Th17 cells among T cells cultured for 3 d in the presence of TGF-β and IL-6. Data are representative of three independent experiments ( a , c , e , g ) or are pooled from three independent experiments ( b , d , f , h ). Summary data are presented as mean ± s.d. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; by unpaired two-tailed Student’s t-tests. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Isolation, Cell Culture, Cell Differentiation, Two Tailed Test

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: C57BL/6 mice were administered IMQ cream on a 2.5 cm × 2.5 cm patch of shaved back skin daily for 7 consecutive days, and were injected with CPT-11 or PBS intraperitoneally once per day ( n = 12 mice per group). a Statistical analysis of epidermal thickness. b Representative histological skin images. c – k Bar graphs showing frequencies of Ki67 + CD4 + T cells ( c ), Ki67 + CD8 + T cells ( d ), IL-17 + CD4 + Th17 cells ( e ), RORγt + CD4 + Th17 cells ( f ), IFN-γ + CD4 + Th1 cells ( g ), T-bet + CD4 + Th1 cells ( h ), IFN-γ + CD8 + T cells ( i ), IL-4 + CD4 + Th2 cells ( j ) and FoxP3 + CD4 + Treg cells ( k ) in the spleen (SPL) and draining lymph nodes (DLN) of indicated groups. Data are representative of three independent experiments ( a , b ) or are pooled from three independent experiments ( c – k ). Summary data are presented as mean ± s.d. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; by a one-way analysis of variance (ANOVA) with Tukey’s post hoc test. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Cream, Injection

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: C57BL/6 mice were subcutaneously immunized with MOG peptide 35–55 emulsified in complete Freund’s adjuvant to induce EAE, and treated with CPT-11 or PBS daily from day 9. a EAE clinical scores of the indicated groups ( n = 10 mice per group). b Representative Luxol Fast Blue (LFB) staining of cervical spinal cord sections. c Representative histological images of cervical spinal cord sections. d , e Representative FACS plots ( d ) and bar graph ( e ) showing frequencies of CD3 + T cells in the brain and spinal cord. f , g Representative FACS plots ( f ) and bar graph ( g ) showing frequencies of IFN-γ + CD4 + Th1 cells in brain and spinal cord. h – k Representative FACS plots ( h , j ) and bar graphs ( l , k ) showing frequencies of Th17 cells in brain and spinal cord. l Bar graph showing frequencies of Foxp3 + Treg cells in brain and spinal cord. Data are representative of two independent experiments ( a – c ) or are pooled from two independent experiments ( d – l ). Summary data are presented as mean ± s.d. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; by unpaired two-tailed Student’s t-tests. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Adjuvant, Staining, Two Tailed Test

Journal: Cell Death Discovery

Article Title: CPT-11 mitigates autoimmune diseases by suppressing effector T cells without affecting long-term anti-tumor immunity

doi: 10.1038/s41420-024-01983-8

Figure Lengend Snippet: C57BL/6 mice were administered IMQ cream on shaved 2.5 cm × 2.5 cm patches of back skin daily for 7 consecutive days and were injected with CPT-11 or PBS intraperitoneally once per day. Approximately 5 weeks after psoriasis induction and treatment, the mice were injected with B16 cells to establish a tumor-bearing model ( n = 7 mice per group). a Experimental scheme of the B16 tumor-bearing model after psoriasis induction and treatment. b Tumor growth curves. c – j Representative FACS plots ( c , e , g , i ) and Bar graphs ( d , f , h , j ) showing frequencies of Ki67 + CD4 + T cells ( c , d ), IFN-γ + CD4 + Th1 cells ( e , f ), IFN-γ + CD8 + cells ( g , h ), and FoxP3 + CD4 + Treg cells ( i , j ). Data are representative of two independent experiments ( b – d ) or are pooled from two independent experiments ( e – j ). Summary data are presented as mean ± s.d. * p < 0.05, ** p < 0.01, **** p < 0.0001; by unpaired two-tailed Student’s t-tests. See also Supplementary Fig. .

Article Snippet: The following chemicals were purchased from the indicated manufacturers: purified anti-mouse CD3 (145–2C11, Bio X Cell, # BE0001–1), purified anti-mouse CD28 (37.51, Bio X Cell, # BE0015–1), recombinant mouse IL-12 (R&D Systems, #419-ML-500), Freund’s adjuvant, incomplete (IFA) (BD/Difco Laboratories, # 263910), Mycobacterium tuberculosis (BD Biosciences, #231141), DNase I (Millipore Sigma, # DN25), Collagenase IV (Thermo Fisher Scientific, # # 17104–019), PMA (Millipore Sigma, #P8139), Ionomycin calcium salt (Millipore Sigma, #13909), Golgi-Plug Protein Transport Inhibitor (BD Biosciences, #555029), CD4 + CD62L + T cell Isolation Kit, mouse (Miltenyi Biotec, #130–106-643), Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific, #00–5523-00), Cytofix/Cytoperm Fixation/ Permeabilization Solution Kit (BD Biosciences, #554714), CellTraceTM CFSE Cell Proliferation Kit (Thermo Fisher Scientific, # C34554), Seahorse XF Cell Mito Stress Test Kit (Agilent, # 103015–100), fluorochrome-conjugated antibodies (zombie yellow [Biolegend, #423104]), anti-mouse CD45 (30-f11, Thermo Fisher Scientific, #56-0451-82), anti-mouse TCRβ (H57-597, Thermo Fisher Scientific, # 47–5961-82), anti-mouse CD4 (RM4-5, Thermo Fisher Scientific, #45-0042-82), anti-mouse CD8α (53-6.7, Thermo Fisher Scientific, #11-0081-85), anti-mouse CD8β (H35-17.2, Thermo Fisher Scientific, 11-008S-85), anti-mouse CD62L (MEL-14, Thermo Fisher Scientific, #11-0621-85), anti-mouse CD44 (IM7, Thermo Fisher Scientific, #17-0441-83), anti-mouse CD25 (PC61.5- Thermo Fisher Scientific, #45-0251-82), anti-mouse CD69 (H1.2F3, Thermo Fisher Scientific, #12-0691-83), anti-mouse IFN-γ (XMG1.2, Thermo Fisher Scientific, #48-7S11-82), anti-mouse IL-17(eBio17B7, Thermo Fisher Scientific, #25-7177-82), anti-mouse IL-4 (11B11, Thermo Fisher Scientific, #25-7041-82), anti-mouse Ki67 (SolA15, Thermo Fisher Scientific, #25-5698-82), anti-mouse RORγt (B2D, Thermo Fisher Scientific, #17-6981-82), anti-mouse T-bet (eBio4B10, Thermo Fisher Scientific, #12-5825-82), anti-mouse FoxP3 (FJK-16s, Thermo Fisher Scientific, #48-577S-82), 7-AAD Viability Staining Solution (Thermo Fisher Scientific, #00-6993-50), and Annexin V (Thermo Fisher Scientific, #BMS306PE-100), anti-mouse CD16/32 (Biolegend, #101302).

Techniques: Cream, Injection, Two Tailed Test