recombinant human cytokines il-2  (PeproTech)

Journal: bioRxiv

Article Title: A Transient Immunostimulatory Niche Synergizes Adoptive and Endogenous Immunity for Enhanced Tumor Control

doi: 10.1101/2025.08.22.671875

Figure Lengend Snippet: ( A ) Schematic illustration demonstrating proposed mechanism of ACTIVATE: (i-ii) Injected hydrogel containing adoptive T cells and cytokine(s) enables long-term local retention of cytokine alongside sustained adoptive T cell expansion and release over the course of 2 weeks; (iii) recruitment of endogenous immune cells, generating a local inflammatory niche for activation and expansion of endogenous immune cells; (iv) adoptive T cell and cytokine release elicit potent tumor cell killing, releasing antigens that can be (v) picked up by antigen-presenting cells and taken back to the tumor-draining lymph nodes for (vi) cross-presentation to endogenous T cells; (vii) combined with the activation and differentiation of endogenous immune cells in the tumor- draining lymph nodes provoked by drainage of adoptive T cells and cytokine enable (viii) engagement of endogenous immune cells for enhanced anti-tumor efficacy. ( B ) Polymer- nanoparticle hydrogels formed through self-assembly of dodecyl-modified hydroxypropyl methylcellulose (HPMC-C12) and degradable PEG-PLA core-shell nanoparticles, enabling co- encapsulation of adoptive T cells and stimulatory cytokines.

Article Snippet: Human cytokines were purchased from R&D Systems (Recombinant IL-2 Protein, Recombinant IL-7 Protein, Recombinant IL-2 Protein, Recombinant IL-15 Protein) and murine cytokines were purchased from Sino Biological (IL-2, IL-7, and IL-12A & IL-12B Heterodimer Protein (His Tag)) and PeproTech (Recombinant murine IL-15).

Techniques: Injection, Activation Assay, Polymer, Modification, Encapsulation

Journal: bioRxiv

Article Title: A Transient Immunostimulatory Niche Synergizes Adoptive and Endogenous Immunity for Enhanced Tumor Control

doi: 10.1101/2025.08.22.671875

Figure Lengend Snippet: ( A ) Schematic of in vitro release assay of cytokines from ACTIVATE immersed in PBS. ( B ) %Mass of cytokines IL-12, IL-7, IL-2, and IL-15 released and remaining in the hydrogels over the 1-week assay. ( C ) Schematic of in vitro cellular release assay: Hydrogel containing PMEL adoptive T cells and cytokine is injected into a porous transwell suspended over medium. Cells proliferate within the hydrogel while also releasing into the medium below. At each timepoint, the medium below the transwell is collected, cells are counted, and transwell is placed into fresh media. ( D ) Cumulative number of cells released into the medium over various timepoints for PMEL and cytokine encapsulated hydrogels and PMEL only control. ( E ) Total number of cells remaining in ACTIVATE on the final day of the assay. ( F ) Total cell exposure determined as the sum of cumulative cells released and total cells remaining in the hydrogel on day 8. ( G ) Cell viability of the cells remaining in the hydrogels on the final day of the assay. ( H ) Representative flow cytometry plots and quantification of PD-1 MFI of PMEL cells in the gel 3 days after encapsulation. ( I ) Frequency of central memory (CD62L + CD44 + ) and effector memory (CD62L - CD44 + ) phenotypes for PMEL cells in the hydrogel 3 days after co-encapsulation with different cytokines. Data are representative of n = 3 technical replicates per group and presented as mean ± s.e.m. P values were determined by ordinary one-way ANOVA followed by Tukey’s multiple comparison test using GraphPad PRISM.

Article Snippet: Human cytokines were purchased from R&D Systems (Recombinant IL-2 Protein, Recombinant IL-7 Protein, Recombinant IL-2 Protein, Recombinant IL-15 Protein) and murine cytokines were purchased from Sino Biological (IL-2, IL-7, and IL-12A & IL-12B Heterodimer Protein (His Tag)) and PeproTech (Recombinant murine IL-15).

Techniques: In Vitro, Release Assay, Injection, Control, Flow Cytometry, Encapsulation, Comparison

Journal: Cell Reports Medicine

Article Title: Coupling IL-2 with IL-10 to mitigate toxicity and enhance antitumor immunity

doi: 10.1016/j.xcrm.2025.102257

Figure Lengend Snippet: IL-10 controls IL-2-mediated induction of inflammatory cytokines PBMCs were cultured with a titration of IL-2, IL-10, or IL-2+IL-10 (0–13 nM) for 24 h followed by measurement of supernatant analytes either before (A and B) or following anti-CD3 (C and D). Statistical analyses are depicted between IL-2 and IL-2+IL-10 groups at the highest concentration (13 nM) by donor-matched pairwise t test (∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001). Results are reported as the mean ± SEM.

Article Snippet: In human primary cell studies, recombinant human cytokines including IL-2 and IL-10 purchased from R&D Systems were used, and recombinant human IFNγ from MedChemExpress.

Techniques: Cell Culture, Titration, Concentration Assay

Journal: Cell Reports Medicine

Article Title: Coupling IL-2 with IL-10 to mitigate toxicity and enhance antitumor immunity

doi: 10.1016/j.xcrm.2025.102257

Figure Lengend Snippet: IFNγ- and TNF-α-dependent IL-2 toxicity is ameliorated by IL-10 (A–D) PBMCs were untreated (control) or cultured with cytokines (1.3 nM) and receptor-blocking antibodies as follows: IL-2, IL-10, IL-2+anti-IFNγR1 (5 μg/mL), or IL-2+anti-IL-10RA (30 μg/mL) for up to 48 h, and supernatant analytes were measured. Data are representative of 5–9 independent donor experiments. Statistical analyses performed by donor pairwise t test on samples collected at 48 h. Statistically significant differences reported between IL-2, IL-2+anti-IFNγR1, and IL-2+anti-IL-10RA ( p value; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001). (E–G) PBMCs were cultured with IL-10 (1.3 nM), IFNγ (10 ng/mL), and IL-10+IFNγ or untreated (control), and supernatant analytes were measured. Data are representative of 4 independent donor experiments. Statistical analyses of the 48 h results were performed by pairwise t test ( p value; ns, not significant; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001). (H) Monocytes were stimulated for up to 16 h with IFNγ alone or with IL-10 (1.3 nM) and stained for IRF1 and IRF8 protein levels. Reported is the median fluorescence intensity (MFI) of IFNγ+IL-10-treated cells relative to the IFNγ-treated cells. Data are representative of 3 independent donor experiments. Statistical analyses performed by one-sample t test (ns, not significant; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01). (I and J) Quantitation of Evans blue dye extravasation into the lungs after IL-2 treatment of wild-type, IFNγ-knockout, and TNF-α-knockout animals (I) and wild-type mice administered anti-TNF-α or an isotype control (J). Data are representative of 6–9 animals per group. Statistical analysis by one-way ANOVA with Tukey’s multiple comparisons test (ns, not significant; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001). Results are reported as the mean ± SEM.

Article Snippet: In human primary cell studies, recombinant human cytokines including IL-2 and IL-10 purchased from R&D Systems were used, and recombinant human IFNγ from MedChemExpress.

Techniques: Control, Cell Culture, Blocking Assay, Staining, Fluorescence, Quantitation Assay, Knock-Out

Journal: Cell Reports Medicine

Article Title: Coupling IL-2 with IL-10 to mitigate toxicity and enhance antitumor immunity

doi: 10.1016/j.xcrm.2025.102257

Figure Lengend Snippet: Coupling and targeting of IL-2 and IL-10 exerts broad immune activation in non-human primates and shows a strong safety profile in a GLP study (A–L) Male, and female, cynomolgus monkeys were treated with DK2 10 (EGFR) (0–2.5 mg/kg) dosed subcutaneously three times a week for 27 days. Each group had both males and females (n = 6–10). (A) DK2 10 (EGFR) plasma concentration plotted up to 72 h after the first dose. (B–E) Plasma cytokines (pg/mL) from animals at the indicated study time points. Concentrations below the lower limit of quantification (LLOQ) are reported as 150 pg/mL. (F) Hematology analyses, represented as fold change, comparing week −1 (pre-dose) to day 28 (post-dose) cell concentrations. Reported are eosinophils, platelets, red blood cells (RBCs), and white blood cells (WBCs). (G) Clinical chemistry on day 28. Reported are alanine transaminase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), hemoglobin (HGB), red blood cell distribution width (RDW), and cholesterol. (H–L) Blood was collected at the indicated times (time from first dose and the most recent dose are indicated on the x axis) to determine immune cell subset frequencies. Subsets reported are total T cells among CD45 + lymphocytes (H), frequency of proliferating (Ki67 + ) Tregs (CD3 + /CD4 + /CD127 low /CD25 + ) (I), CD4 + T cells (J), CTLs (K), and the ratio of CTLs to Tregs (L). Statistical analyses were performed by one-way ANOVA with Tukey’s multiple comparisons test on measurement at the final time point. Results are reported as the mean ± SD. LLOQ is indicated on several plots.

Article Snippet: In human primary cell studies, recombinant human cytokines including IL-2 and IL-10 purchased from R&D Systems were used, and recombinant human IFNγ from MedChemExpress.

Techniques: Activation Assay, Clinical Proteomics, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Subtype-specific atypical B cell profiles in myasthenia gravis reveal distinct immunopathological pathways

doi: 10.3389/fimmu.2025.1608160

Figure Lengend Snippet: CD11c + B cells differentiate into ASCs and produce autoantibodies in MuSK-MG. Isolated CD11c − and CD11c + B cells were stimulated with CD40L, IL-2, IL-12, and BAFF (n = 6 for each group) for 14 days. AChR-MG subjects were stratified based on disease onset (LOMG n = 2, EOMG n = 4). In LPS-treated wells, cells were stimulated with CD40L and cytokines (IL-2, IL-4, IL-21, and BAFF) for 14 days, with LPS added 48 hours prior to harvest (n = 3). (A) Representative flow cytometry plots of CD11c + B cells from a MuSK-MG subject. Antibody-secreting cells (ASCs) were identified as CD27 ++ CD38 ++ cells. (B) Quantification of CD11c − and CD11c + B cells that differentiated into + CD27 ++ CD38 ++ ASCs. (C) Total IgG production (ng/mL) in culture supernatants as measured by ELISA. (D) Radioimmunoassay measuring anti-AChR and anti-MuSK antibody levels in culture supernatants from CD11c + B cells (fmol/mL) stimulated with CD40L, IL-2, IL-4, IL-21 and BAFF for 14 days. Data are represented as mean ± SEM.

Article Snippet: 2x10 3 cells were plated with wells seeded with 3 x10 3 MS-40L lo cells (kindly provided by the Kelsoe Laboratory) per well and cultured in complete RPMI media in the presence of recombinant human cytokines IL-2 (50 ng/mL), IL-4 (10 ng/mL), IL-21(10 ng/mL) and BAFF (10 ng/mL) (all purchased from Peprotech) for 14 days (n = 6 per group).

Techniques: Isolation, Flow Cytometry, Enzyme-linked Immunosorbent Assay, RIA Assay