Journal: Biomedicines

Article Title: IL-15, IL-18 and IL-21 Along the Stress–Smoking–Periodontal Health Axis: A Cross-Sectional Study in Mexican Adults

doi: 10.3390/biomedicines14010114

Figure Lengend Snippet: Serum cytokines by periodontal status (binary). ( A ) IL-15, ( B ) IL-18, and ( C ) IL-21 concentrations in healthy vs. periodontal disease (gingivitis + periodontitis). Violin plots display distribution density; overlaid boxplots show median and IQR; points represent individual participants. Y-axes are shown on a logarithmic scale to improve readability of right-skewed distributions.

Article Snippet: IL-18, IL-21, and IL-15 were quantified in serum using commercial antibody-based ELISA kits: R&D Systems DuoSet ® Human IL-15 (DY247), R&D Systems DuoSet ® Human Total IL-18 (DY318) (R&D Systems, Inc., Minneapolis, MN, USA), and BioLegend ELISA MAXTM Deluxe Set Human IL-21 (433804) (BioLegend, San Diego, CA, USA).

Techniques:

Journal: Journal of Extracellular Vesicles

Article Title: Endothelium‐Treg Communication Through Extracellular Vesicle Transfer Exacerbates Acute Respiratory Distress Syndrome

doi: 10.1002/jev2.70235

Figure Lengend Snippet: LPS‐EC‐EVs initiate IL‐21 production in Tregs. (a) Volcano plot of RNA‐seq expression in Tregs exposed to EC‐EVs or LPS‐EC‐EVs for 5 days. Orange dots indicated upregulated genes (fold change > 2, p value < 0.05), whereas green dots represented downregulated ones (fold change < 0.5, p value < 0.05) in Tregs with LPS‐EC‐EVs. (b) Graphic overview of the upregulated genes by IPA. (c) The IL‐21 centered molecular network generated by IPA. (d) RT‐qPCR combined ELISA analyses of IL‐21 expressions in Tregs exposed to either PBS or different EVs for 5 days ( n = 3). (e) FCM and (f) quantification of IL‐21 staining in Tregs treated with either PBS or different EVs ( n = 3). (g) Representative images of immunofluorescence staining of IL‐21 expression in Tregs in vitro . Naïve CD4 + T cells from Foxp3 YFP/Cre were polarized into Tregs exposed to either PBS or different EVs; Foxp3 (green), IL‐21 (red), Scale bar, 3 µm. (h) Representative confocal views and (i) Immunoblotting showing STAT3 activation exposed to LPS‐EC‐EVs ( n = 3). In (h), Scale bar, 4 µm. * p < 0.05, *** p < 0.005, **** p < 0.001. See also Figure .

Article Snippet: Supernatants were collected at Day 5, and the concentration of TGF‐β1 (DB100C, R&D, USA), KGF (ELK3081, ELK Biotech, China), IFN‐γ (E‐EL‐M0048, Elabscience, China), IL‐4 (E‐EL‐M0043, Elabscience, China), IL‐17A (E‐EL‐M0047, Elabscience, China) and IL‐21 (EK0797, BOSTER, China) were determined by ELISA assays, according to the respective manufacturers’ instructions.

Techniques: RNA Sequencing, Expressing, Generated, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence, In Vitro, Western Blot, Activation Assay

Journal: Journal of Extracellular Vesicles

Article Title: Endothelium‐Treg Communication Through Extracellular Vesicle Transfer Exacerbates Acute Respiratory Distress Syndrome

doi: 10.1002/jev2.70235

Figure Lengend Snippet: IL‐21 signalling blockade ameliorates the EV‐induced Treg suppression. aIL‐21 (a), aIL‐21R (b) and Stattic (c) were seen to rescue EV‐induced Treg suppression in a dose‐dependent manner ( n = 3). (d) Immunoblotting of the phosphor‐STAT3 and STAT3 levels of Tregs exposed to LPS‐EC‐EVs after treatment with IL‐21 signalling inhibitors for 5 days ( n = 3). (e and f) Splenic naïve CD4 + T cells were isolated from WT and Il21 −/− mice and then polarized to Tregs with LPS‐EC‐EVs for 5 days. The percentage of Tregs were analyzed via FCM (e) and data were statistically analyzed (f), ( n = 6). (g) Schematic diagram of ALI induction in chimeras (created using Biorender.com). (h) Quantification of Tregs in BALF and BLNs from different chimeras via FCM ( n = 5). (i) Representative HE stained tissue sections and lung injury score evaluation ( n = 5 in each group). Scale bar, 100 µm. (j) Immunofluorescence staining and quantitative analysis of MPO ( n = 5 in each group). Scale bar, 100 µm. (k and l) Immunohistochemical staining and quantitative analysis of Ly6G (k) and NE (l) ( n = 5 in each group). Scale bar, 100 µm. ** p < 0.01, *** p < 0.005, **** p < 0.001. See also Figure .

Article Snippet: Supernatants were collected at Day 5, and the concentration of TGF‐β1 (DB100C, R&D, USA), KGF (ELK3081, ELK Biotech, China), IFN‐γ (E‐EL‐M0048, Elabscience, China), IL‐4 (E‐EL‐M0043, Elabscience, China), IL‐17A (E‐EL‐M0047, Elabscience, China) and IL‐21 (EK0797, BOSTER, China) were determined by ELISA assays, according to the respective manufacturers’ instructions.

Techniques: Western Blot, Isolation, Staining, Immunofluorescence, Immunohistochemical staining

Journal: Journal of Extracellular Vesicles

Article Title: Endothelium‐Treg Communication Through Extracellular Vesicle Transfer Exacerbates Acute Respiratory Distress Syndrome

doi: 10.1002/jev2.70235

Figure Lengend Snippet: Med1 in LPS‐EC‐EVs serves as an IL‐21 activating TF. (a) Venn diagram of upregulated proteins in LPS‐EC‐EVs and IL‐21 predicted TFs showing that only Med1 was accumulated. (b) Immunoblotting combined with Ponceau S staining confirmed that Med1 was selectively enriched in LPS‐EC‐EVs rather than EC‐EVs ( n = 3). (c) Representative IEM image showing the presence of Med1 in LPS‐EC‐EVs. Scale bar, 100 nm. (d) 3D view indicated the GFP‐labelled EVs in the Treg nucleus (blue) after coculturing. (e) Immunoblotting analysis confirmed that Med1 was transferred into the Treg nucleus through EVs. Tregs cocultured with Flag‐Med1 LPS‐EC‐EVs and proceeded to isolate nuclear and cytoplasmic fractions ( n = 3). (f) Schematic model of the primer targeting sites (left) and Flag‐tagged ChIP‐qPCR analysis (right) demonstrated attachment of Med1 to the predicted region on IL‐21 ( n = 5). (g) Gel‐based imaging of Flag‐tagged ChIP‐qPCR products from (f). (h) Dual‐luciferase assays were performed to confirm the interaction between Med1 and IL‐21 ( n = 6). *** p < 0.005, **** p < 0.001. See also Figures and .

Article Snippet: Supernatants were collected at Day 5, and the concentration of TGF‐β1 (DB100C, R&D, USA), KGF (ELK3081, ELK Biotech, China), IFN‐γ (E‐EL‐M0048, Elabscience, China), IL‐4 (E‐EL‐M0043, Elabscience, China), IL‐17A (E‐EL‐M0047, Elabscience, China) and IL‐21 (EK0797, BOSTER, China) were determined by ELISA assays, according to the respective manufacturers’ instructions.

Techniques: Western Blot, Staining, ChIP-qPCR, Imaging, Luciferase

Journal: Journal of Extracellular Vesicles

Article Title: Endothelium‐Treg Communication Through Extracellular Vesicle Transfer Exacerbates Acute Respiratory Distress Syndrome

doi: 10.1002/jev2.70235

Figure Lengend Snippet: Med1 in LPS‐EC‐EVs is the key modulator for Treg restriction. (a) Immunoblotting showed the Med1 expression in WT and Med1 knockdown ECs ( n = 3). (b) Med1 depletion efficiency in EVs from WT and Med1 knockdown ECs ( n = 3). FCM (c) and quantification (d) of CD25 + Foxp3 + double staining in Tregs exposed to Med1‐KD LPS‐EC‐EVs ( n = 3). (e) RT‐qPCR (left) and ELISA (right) of IL‐21 expression in Tregs and cell culture supernatants ( n = 3). (f) Seven‐day survival analysis of LPS‐IT mice in shMed1‐AAV‐LungX group and shNC‐AAV‐LungX group. Immunoblotting (g) and quantification (h) of Med 1 expression in BALF‐EVs after AAV‐shNC or AAV‐shMed1 infection ( n = 5). (i) Quantification of Tregs in BALF and BLNs from different group mice via FCM at 72 h after LPS administration ( n = 5). (j) Representative HE stained tissue sections and lung injury score evaluation ( n = 5 in each group). Scale bar, 100 µm. (k) Immunofluorescence staining and quantitative analysis of MPO ( n = 5 each group). Scale bar, 100 µm. (l and m) Immunohistochemical staining and quantitative analysis of Ly6G (l) and NE (m) ( n = 5 each group). Scale bar, 100 µm. * p <0.05, ** p < 0.01, *** p <0.005, **** p < 0.001. See also Figure .

Article Snippet: Supernatants were collected at Day 5, and the concentration of TGF‐β1 (DB100C, R&D, USA), KGF (ELK3081, ELK Biotech, China), IFN‐γ (E‐EL‐M0048, Elabscience, China), IL‐4 (E‐EL‐M0043, Elabscience, China), IL‐17A (E‐EL‐M0047, Elabscience, China) and IL‐21 (EK0797, BOSTER, China) were determined by ELISA assays, according to the respective manufacturers’ instructions.

Techniques: Western Blot, Expressing, Knockdown, Double Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Infection, Staining, Immunofluorescence, Immunohistochemical staining

Journal: Journal of Extracellular Vesicles

Article Title: Endothelium‐Treg Communication Through Extracellular Vesicle Transfer Exacerbates Acute Respiratory Distress Syndrome

doi: 10.1002/jev2.70235

Figure Lengend Snippet: Elevated Med1 expression in BALF‐EVs from ARDS patients. (a) Comparison of CD31 + EVs percentage in the BALF from non‐ARDS ( n = 5) and ARDS ( n = 9) patients. Immunoblotting (b) and quantification (c) of Med1 expression in BALF‐EVs from non‐ARDS ( n = 5) and ARDS patients ( n = 9). (d) IL‐21 concentrations in the BALF from non‐ARDS ( n = 5) and ARDS ( n = 9) patients. (e) Correlation analysis of different variables including CD31 + EV proportion, Med1 relative expression in BALF‐EVs and IL‐21 concentration in BALF. * p < 0.05, ** p < 0.01, **** p < 0.001. See also Figure .

Article Snippet: Supernatants were collected at Day 5, and the concentration of TGF‐β1 (DB100C, R&D, USA), KGF (ELK3081, ELK Biotech, China), IFN‐γ (E‐EL‐M0048, Elabscience, China), IL‐4 (E‐EL‐M0043, Elabscience, China), IL‐17A (E‐EL‐M0047, Elabscience, China) and IL‐21 (EK0797, BOSTER, China) were determined by ELISA assays, according to the respective manufacturers’ instructions.

Techniques: Expressing, Comparison, Western Blot, Concentration Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: Construction and characterization of IL-21-secreting Salmonella . (A) Plasmid map designed for the secretion of FlgM-tagged IL-21. (B) After overnight culturing of the Salmonella strain secreting IL-21, 0.2% l -arabinose was added to induce IL-21 expression. Whole cells, cell lysates, and supernatants were collected, and Western blot analysis was performed using anti-His and anti-IL-21 antibodies, respectively. (C) IL-21 secretion was confirmed by ELISA ( n = 5). (D) Growth of the strain secreting IL-21 was compared between induced (0.2% l -arabinose) and non-induced groups by measuring the OD at 600 nm. (E) Western blot analysis using anti-His was conducted to track the secretion of IL-21 over time following induction with 0.2% l -arabinose. (F) Motility assays were performed to examine the motility of the IL-21-secreting strain under 0.2% l -arabinose. The experiments were repeated at least three times, and representative data are shown. Scale bar = 5 mm. Significance is indicated as ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: Plasmid Preparation, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: Antitumor efficacy of IL-21-secreting Salmonella in tumor-bearing mice. (A) Treatment schedule for cancer therapy using IL-21-secreting Salmonella . CT26 cells (5 × 10 5 ) were subcutaneously injected into the right flank of BALB/c mice. When the tumor size reached approximately 100 mm 3 , Salmonella (2 × 10 7 CFU) was administered intravenously, and from Day 3 post-injection, l -arabinose (80 mg/mouse) was administered intraperitoneally. IL-21(−), without l -arabinose; IL-21(+), with l -arabinose. (B) Representative images of tumor-bearing mice. (C) Tumor volume at 2-day intervals. (D) Body weight changes in each treatment group. (E) Survival curves for each treatment group (data from B–E; n = 7). (F) TUNEL analysis of tumors from each treatment group. (G) Ki-67 expression in tumors from each treatment group. bar = 50 μm. Image quantification data are presented in (H, I) (data from F–I; n = 3). Significance is indicated as ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Data are presented as the mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: Injection, TUNEL Assay, Expressing

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: IL-21-secreting Salmonella promotes T cell infiltration and antitumor activity. (A) Experimental scheme in tumor-bearing mice: Immune cells were analyzed in tumor tissues on Day 7. (B) Anti-IL-21 was detected in the tumor tissues using ELISA. (C) In tumor tissues, anti-IL-21 was confirmed through IHC. Scale bar = 50 μm. (D) Image quantification data (data from B–D; n = 3). (E–G) Flow cytometry analysis showed that CD4 + and CD8 + T cells were activated by IL-21 in mouse tumor tissues (data from E–G; n = 3). Significance is indicated as ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Data are presented as the mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: IL-21-secreting Salmonella promotes T cell infiltration and antitumor activity. The schedule for immune cell analysis is shown in A. (A, B) Tumor tissues obtained on Day 7 were fixed and processed for IF analysis. CD4 + and CD8 + T cells were analyzed using IF staining. (C, D) Granzyme B and Perforin levels in CD8 + T cells were analyzed using immunofluorescence staining. Scale bar = 50 μm. Image quantification data are presented in (E–H) ( n = 3). Significance is indicated by ∗ P < 0.05, ∗∗ P < 0.01, ns = not significant. Data are presented as the mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: Activity Assay, Cell Analysis, Staining, Immunofluorescence

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: Antitumor efficacy of IL-21-secreting Salmonella combined with anti-PD-L1 antibody in tumor-bearing mice. (A) Schedule for combination therapy with IL-21-secreting Salmonella and anti-PD-L1 antibody. CT26 cells (5 × 10 5 ) were subcutaneously injected into the right flank of BALB/c mice. When the tumor size reached approximately 100 mm 3 , Salmonella (2 × 10 7 CFU) was administered intravenously, and simultaneously, anti-PD-L1 antibody (200 μg/mouse) was administered intraperitoneally four times starting from Day 0. Additionally, starting at 3 dpi, l -arabinose (80 mg/mouse) was administered daily via the intraperitoneal route. IL-21(−), without l -arabinose; IL-21(+), with l -arabinose. (B) Representative images of tumor-bearing mice. (C) Tumor volume at 2-day intervals. (D) Body weight changes for each treatment group. (E) Survival curves for each treatment group (data from B–E; n = 5). (F) TUNEL analysis of tumors from each treatment group. (G) Ki-67 expression images of tumors from each treatment group. Scale bar = 50 μm. Image quantification data are presented in (H, I) (data from F–I; n = 3). Significance is indicated at ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001; ns = not significant. Data are presented as the mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: Injection, TUNEL Assay, Expressing

Journal: Acta Pharmaceutica Sinica. B

Article Title: Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects

doi: 10.1016/j.apsb.2025.09.025

Figure Lengend Snippet: IL-21-secreting Salmonella exhibits in vivo biosafety. (A) IL-21-secreting Salmonella (2 × 10 7 CFU) was administered intravenously, and observations were made at 2, 24, 48, 72, 168, and 336 h. (B) After Salmonella administration as described in (A), tissue homogenates of the heart, liver, spleen, lung, kidney, and tumor were cultured on solid LB agar plates and photographed on Days 1, 7, and 14. (C) Time-dependent quantitative distribution of Salmonella in tumors and major organs in each group (data from A; n = 3). (D) Number of IL-21-secreting Salmonella clones in major organs and tumors (Data from B; n = 3). Data are presented as mean ± standard deviation (E–H). ALT, AST, BUN, and CREA biochemical indicators were measured in the serum at Days 7, 14 and 21 post-administrations. The yellow-shaded areas indicate the average normal range ( n = 3). Data are presented as the mean ± SEM.

Article Snippet: After collecting all supernatant samples, the IL-21 concentration was measured using a mouse IL-21 ELISA Kit (KE10012, Proteintech, Rosemont, IL, USA).

Techniques: In Vivo, Cell Culture, Clone Assay, Standard Deviation