Journal: Theranostics

Article Title: Human umbilical cord-derived mesenchymal stem cell therapy ameliorates lupus through increasing CD4+ T cell senescence via MiR-199a-5p/Sirt1/p53 axis

doi: 10.7150/thno.48080

Figure Lengend Snippet: hUC-MSCs treatment normalizes markers of senescence in MRL/ lpr mice splenic CD4+ T cells in vivo. (A, B) qPCR analysis showing the expression levels of p21, p16 in WT mice, PBS-treated MRL/ lpr mice and hUC-MSCs-treated MRL/ lpr mice. (C) Representative western blot showing the expression levels of p21, p16, p53 and acetylation of p53 in WT mice, PBS-treated MRL/ lpr mice and hUC-MSCs-treated MRL/ lpr mice. (D, E) Capillary WES and qPCR analysis showing the expression of Sirt1 in WT mice, PBS-treated MRL/ lpr mice and hUC-MSCs-treated MRL/ lpr mice. GAPDH was used as a protein loading control. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The splenic CD4+ T cells were purified using immunomagnetic positive selection (EasySepTM mouse CD4+ positive selection Kit, 18952, Stemcell technologies, Canada).

Techniques: In Vivo, Expressing, Western Blot, Control

Journal: Theranostics

Article Title: Human umbilical cord-derived mesenchymal stem cell therapy ameliorates lupus through increasing CD4+ T cell senescence via MiR-199a-5p/Sirt1/p53 axis

doi: 10.7150/thno.48080

Figure Lengend Snippet: hUC-MSCs increase markers of senescence in splenic CD4+ T cells in vitro . (A-C) Splenic CD4+ T cells from MRL /lpr mice were cultured alone or with hUC-MSCs at ratios of 1:1, 10:1, or 50:1 for 48 h. (D-F) In further experiments, MRL/ lpr splenic CD4+ T cells and hUC-MSCs (10:1) were cultured separated by transwells. Sirt1 (A, D), p21 (B, E) and p16 (C, F) RNA levels of CD4+ T cells were quantitated by qPCR. All experimental data were verified in at least two independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; n.s., not significant.

Article Snippet: The splenic CD4+ T cells were purified using immunomagnetic positive selection (EasySepTM mouse CD4+ positive selection Kit, 18952, Stemcell technologies, Canada).

Techniques: In Vitro, Cell Culture

Journal: Theranostics

Article Title: Human umbilical cord-derived mesenchymal stem cell therapy ameliorates lupus through increasing CD4+ T cell senescence via MiR-199a-5p/Sirt1/p53 axis

doi: 10.7150/thno.48080

Figure Lengend Snippet: Sirt1 is a mediator of hUC-MSCs increasing senescence of splenic CD4+ T cells in MRL/ lpr mice. (A) Western blotting showing the expression of Sirt1, p21, p16, p53 and acetylation of p53 in EX527 and DMSO treated MRL/ lpr splenic CD4+ T cells. (B) MRL/ lpr splenic CD4+ T cells were pretreated with SRT1720 12 h before exposed to hUC-MSCs for 24-48 h, western blotting showing the expression of p21, p16, p53 and acetylation of p53. GAPDH was used as a protein loading control. All experimental data were verified in at least three independent experiments. * p < 0.05; ** p < 0.01.

Article Snippet: The splenic CD4+ T cells were purified using immunomagnetic positive selection (EasySepTM mouse CD4+ positive selection Kit, 18952, Stemcell technologies, Canada).

Techniques: Western Blot, Expressing, Control

Journal: Theranostics

Article Title: Human umbilical cord-derived mesenchymal stem cell therapy ameliorates lupus through increasing CD4+ T cell senescence via MiR-199a-5p/Sirt1/p53 axis

doi: 10.7150/thno.48080

Figure Lengend Snippet: hUC-MSCs induced miR-199a-5p can increase MRL/ lpr splenic CD4+ T cell senescence. (A) qPCR analysis of the levels of ten potential miRNAs in WT mice, PBS-treated MRL/ lpr mice and hUC-MSCs-treated MRL/ lpr mice splenic CD4+ T cells. (B-D) qPCR and western blotting analysis of the levels of miR-199a-5p, Sirt1, p21, p16 and acetyl-p53 in vehicle and miR-199a-5p mimic-treated MRL/ lpr splenic CD4+ T cells. (E-F) qPCR and western blotting analysis of the levels of Sirt1, p21, p16 and acetyl-p53 in vehicle and miR-199a-5p inhibitor-treated WT splenic CD4+ T cells. (G-I) MRL/ lpr splenic CD4+ T cells and hUC-MSCs were cultured alone or together in the presence or absence of miR-199a inhibitor using a transwell system. MiR-199a-5p, Sirt1, p21, p16 and acetyl-p53 were quantified in hUC-MSCs (the first bar) or splenic CD4+ T cells (the last three bars). GAPDH was used as a protein loading control. All experimental data were verified in at least two independent experiments. * p < 0.05; ** p < 0.01.

Article Snippet: The splenic CD4+ T cells were purified using immunomagnetic positive selection (EasySepTM mouse CD4+ positive selection Kit, 18952, Stemcell technologies, Canada).

Techniques: Western Blot, Cell Culture, Control

Journal: Theranostics

Article Title: Human umbilical cord-derived mesenchymal stem cell therapy ameliorates lupus through increasing CD4+ T cell senescence via MiR-199a-5p/Sirt1/p53 axis

doi: 10.7150/thno.48080

Figure Lengend Snippet: MiR-199a-5p agomir treatment increases senescence of MRL/ lpr splenic CD4+ T cells. (A) Experimental outline. CD4+ T cells were harvested to measures miR-199a-5p (B), Sirt1 (C), p21 and p16 (D). GAPDH was used as a protein loading control. (n = 6 per group). * p < 0.05; ** p < 0.01.

Article Snippet: The splenic CD4+ T cells were purified using immunomagnetic positive selection (EasySepTM mouse CD4+ positive selection Kit, 18952, Stemcell technologies, Canada).

Techniques: Control

Journal: Cell Death & Disease

Article Title: Colorectal cancer patients-derived immunity-organoid platform unveils cancer-specific tissue markers associated with immunotherapy resistance

doi: 10.1038/s41419-024-07266-5

Figure Lengend Snippet: A Immunohistochemical staining of CD3, CD4, CD8, CD68 and Ki-67 in CRC tissues showed the presence of T-cells populations (CD4 + , CD8 + ) and macrophages (CD68 + ) in MSI and MSS patients. Ki-67 was used as a control proliferation marker of tumor cells; ( B ) Quantification of positive cells/area was performed with QuPath Software. Significance shown refers to Wilcoxon test p -values *< 0.05, **< 0.01, ***< 0.001.

Article Snippet: T-cells and Myeloid Derived Suppressor Cells (MDSCs) were isolated from patient peripheral blood mononuclear cells (PBMCs) by CD3 + immunomagnetic negative selection (#17951, StemCell Technologies) and CD33 + immunomagnetic positive selection (#17876, StemCell Technologies) following positive selection with CD3 + immunomagnetic positive selection (#17851, StemCell Technologies), respectively.

Techniques: Immunohistochemical staining, Staining, Control, Marker, Software

Journal: EBioMedicine

Article Title: Radiotherapy Upregulates Programmed Death Ligand-1 through the Pathways Downstream of Epidermal Growth Factor Receptor in Glioma

doi: 10.1016/j.ebiom.2018.01.027

Figure Lengend Snippet: The effect of PD-L1 blockade on the suppressing ability of tumor cells for CD8 + T cell proliferation. Cell lines (U87 and U251) were treated with RT (5 Gy), PD-L1 Ab (20 μg/mL), RT + PD-L1 Ab, AG490 (10 μM), RT + AG490, or RT + AG490 + PD-L1 Ab, and then co-cultured with CD8 + T cells. At 48 h, CD8 + T cell proliferation was evaluated by FCM. Non-specific Stimulation: anti-CD3/CD28 beads. Representative images and quantitative data are shown. Each column is shown as the means of three separate experiments; bars, SD. Student's t -test; *** P < 0.001. ANOVA; ns, nonsignificant.

Article Snippet: CD8 + T cell immunomagnetic beads positive selection kit was purchased from Stem-Cell Tech (USA).

Techniques: Cell Culture

Journal: EBioMedicine

Article Title: Radiotherapy Upregulates Programmed Death Ligand-1 through the Pathways Downstream of Epidermal Growth Factor Receptor in Glioma

doi: 10.1016/j.ebiom.2018.01.027

Figure Lengend Snippet: The effect of PD-L1 blockade on the cytotoxicity of CD8 + T cells against glioma cells. Cell lines (U87 and U251) were treated with RT (5 Gy), PD-L1 Ab (20 μg/mL), RT + PD-L1 Ab, AG490 (10 μM), RT + AG490, or RT + AG490 + PD-L1 Ab, and then co-cultured with CD8 + T cells. At 48 h, tumor cell apoptosis was evaluated by FCM. Representative images and quantitative data are shown. Each column is shown as the means of three separate experiments; bars, SD. Student's t-test; *** P < 0.001. ANOVA; ns, nonsignificant.

Article Snippet: CD8 + T cell immunomagnetic beads positive selection kit was purchased from Stem-Cell Tech (USA).

Techniques: Cell Culture