Journal: Nature communications

Article Title: Targeting IL-17A enhances imatinib efficacy in Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia.

doi: 10.1038/s41467-023-44270-3

Figure Lengend Snippet: Fig. 5 | IL-17A stimulates the secretion of CXCL16 from leukemia cells, pro- moting the differentiation and migration of Th17 cells. a B220+ cells and Th17 cells in the BM niche in WT mice and BCR-ABLtTA mice (n = 3 mice per group) were subjected to immunofluorescence staining. Representative images were shown. b The numbers of Th17 cells migrating to culture medium or to leukemia cells treated with or without 50 ng/ml rhIL-17A were determined by cell counting and FCS analysis. c Real-time PCR analysis of the relative mRNA levels of chemokine genes in SupB15 (left) and BV173 cells (right) treated with rhIL-17A (100 ng/ml) or PBS control. d The human IL-5, CXCL5, CCL5 and CXCL16 levels in the culture supernatant of SupB15 cells treated with or without rhIL-17A (100 ng/ml) were measured by ELISA. e The CXCL16 concentrations in the serum of BCR-ABLtTA mice and WT mice (n = 5 mice per group) were quantified with a Mouse Chemokine Assay Q1 (Raybiotech). f The CXCL16 concentrations in the serum of HDs (n = 7 samples)

Article Snippet: Three days after transplantation, 70mg/kg imatinib (Sigma–Aldrich, SML1027, p.o., once a day), 5mg/kg anti-IL-17A neutralizing antibody (Bio X Cell, BP0173, i.v., twice a week), or 0.5mg/kg anti-mouse CXCL16 neutralizing antibody (R&D,MAB503, i.v., twice aweek)were administered for 3 consecutive weeks.

Techniques: Migration, Staining, Cell Counting, Real-time Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Traditional and Complementary Medicine

Article Title: Polysaccharides derived from alkali-extracted vinegar-baked Radix Bupleuri suppress hyperimmune T lymphocytes and ameliorate skin graft rejection

doi: 10.1016/j.jtcme.2024.11.006

Figure Lengend Snippet: The effect of KVBCP3 and KVBCP4 on immune cytokines and Treg cells of mice with skin allograft rejection. (A) The secretion of IL-2 in serum was determined by Elisa assay (n = 6). (B) The secretion of IL-4 in serum was determined by Elisa assay (n = 6). (C) Representative flow cytometry plots of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day; (D) The change of the proportions of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day (n = 3). (E) Representative flow cytometry plots of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day; (F) The change of the proportions of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day (n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs Model group, one way ANOVA.

Article Snippet: Cell stimulation and Protein Transport Inhibitor Kit (Lot: AK22752), FITC Anti-Mouse CD4 Antibody (Lot: AF21880), PE Anti-Mouse IL-17A Antibody (Lot: AF17079) were purchased from Elabscience (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 3 | JHU083 inhibited T cells differentiation in vivo. Mice of Vehicle-AIH and JHU083-AIH group were intravenously injected with 8ml/kg ConA, and Vehicle- WT group was injected with normal saline of equal volume. JHU083-AIH group was gavaged with 0.3mg/kg JHU083 24h and 1h before ConA injection, and Vehicle- WT and Vehicle-AIH group were treated with equal vehicle at the same time points. (A) Flow cytometry was applied to analyze the Th1 cell percentage via IFNg expression, the Th2 cell percentage via IL4 expression, and the Th17 cell percentage via IL17A expression. (B) ELISA was applied to analyze serum IFNg (Th1 cells) and IL17 (Th17 cells) secretions. Flow cytometry was also applied to analyze the differentiation of CD8(+) T cells into cytotoxic T lymphocytes, evidenced by secreting IFNg (C) and Granzyme (D). Data were expressed as means ± SEM. *P < 0.05, **P <0.01, and ***P <0.001. ns, not significant.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: In Vivo, Injection, Saline, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 5 | DON treatment suppressed T cells differentiation in vitro. Freshly separated spleen cells were stimulated with 1.5mg/ml ConA with or without treating 2.5mM DON, and cells were analyzed 24h after ConA stimulation. (A) The differentiation markers of CD4+ T cells (IFNg for Th1 cells, IL4 for Th2 cells and IL17 for Th17 cells) were detected using flowcytometry. (B) qRT-PCR was applied to analyze IFN-g, IL4 and IL17 mRNA levels. (C) ELISA was applied to analyze supernatant IFN-g and IL17 secretions. Flow cytometry was also applied to detect the production of activation markers of CTL, such as IFNg (D) and Granzyme B (E). Data were expressed as means ± SEM. **P <0.01 and ***P <0.001, ns, not significant.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Activation Assay

Journal: Frontiers in Immunology

Article Title: VISTA + follicular regulatory T cells modulate the function of effector immune cells: implications for ovarian cancer immune escape

doi: 10.3389/fimmu.2025.1704048

Figure Lengend Snippet: Effects of VISTA + /VISTA − Tfr cells on CD25 − CD4 + T cell differentiation. (A) Flow cytometry scatter plots showing the effects of VISTA + /VISTA − Tfr cells in the co-culture system on the expression of differentiation markers IFN-γ, IL-4, and IL-17A on CD25 − CD4 + T cells (n=3). (B–D) Quantitative bar graphs showing the expression proportions of Th1, Th2, and Th17 cells after co-culture of CD25 − CD4 + T cells with VISTA − Tfr cells or VISTA − Tfr cells overexpressing VISTA (LV-VISTA). (E–G) Quantitative bar graphs showing the expression proportions of Th1, Th2, and Th17 cells after co-culture of CD25 − CD4 + T cells with VISTA + Tfr cells or VISTA + Tfr cells with VISTA silenced (shRNA-VISTA). The statistical results are expressed as the mean ± SD, * p < 0.05, ** p < 0.01, ns, no significant difference.

Article Snippet: Then, 5 μL each of IL-4-APC (MultiSciences, F11IL403), IFN-γ-PE (MultiSciences, F11IFNG02), and IL-17A-FITC (Elabscience, E-AB-F1173C) antibodies were added, followed by incubation at 4°C in the dark for 30 min.

Techniques: Cell Differentiation, Flow Cytometry, Co-Culture Assay, Expressing, shRNA