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: BioMed Research International

Article Title: Chronic Heat Stress Inhibits Immune Responses to H5N1 Vaccination through Regulating CD4 + CD25 + Foxp3 + Tregs

doi: 10.1155/2013/160859

Figure Lengend Snippet: CHS regulates Tregs prevalence. Splenic T cells from each group ( n = 3) were isolated on the indicated days postprimary immunization and then analyzed by flow cytometry. (a) Percentages of CD25 + and Foxp3 + cells in CD4 + T cells were calculated at the indicated time points. Results shown are pooled from three independent repeats. (b) Intracellular staining of IL-10 + CD4 + T cells on day 21 was performed (stimulated with a viral-specific antigen for 6 h in vitro before being analyzed by flow cytometry). Representative plots showing the percentages of positive T cells are shown on the left. Data from all experiments are summarized in bar graphs to the right. (c) The expression levels of TGF- β in the CD4 + cells of CHS groups were analyzed by real-time PCR at indicated time points. Data are presented as relative changes in gene expression (fold) normalized to the TN groups and are one representative from three independent experiments.

Article Snippet: CD4 + CD25 + Tregs were isolated and purified using the MagCellect Mouse CD4 + CD25 + T Cell Isolation Kit according to the manufacturer's protocol (R&D Systems, Inc., Minneapolis, USA).

Techniques: Isolation, Flow Cytometry, Staining, In Vitro, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: BioMed Research International

Article Title: Chronic Heat Stress Inhibits Immune Responses to H5N1 Vaccination through Regulating CD4 + CD25 + Foxp3 + Tregs

doi: 10.1155/2013/160859

Figure Lengend Snippet: Adoptive transfer of CD4 + CD25 + Tregs reduces the vaccine efficacy under TN conditions. 10 6 CD4 + CD25 + Tregs from CHS+Ag group and TN+Ag group were adoptively transferred into TN+Ag mice on day 21 in conjunction with a 50 PFU H5N1 virus challenge. The mortalities of the two transferred groups ( n = 6) were observed for 14 days.

Article Snippet: CD4 + CD25 + Tregs were isolated and purified using the MagCellect Mouse CD4 + CD25 + T Cell Isolation Kit according to the manufacturer's protocol (R&D Systems, Inc., Minneapolis, USA).

Techniques: Adoptive Transfer Assay, Virus