recombinant mouse il12  (PeproTech)

Journal: Molecular Cancer Therapeutics

Article Title: XTX301, a Tumor-Activated Interleukin-12 Has the Potential to Widen the Therapeutic Index of IL12 Treatment for Solid Tumors as Evidenced by Preclinical Studies

doi: 10.1158/1535-7163.MCT-23-0336

Figure Lengend Snippet: Masking of XTX301 activity and reactivation by MMPs in vitro (A) SPR was used to measure the binding kinetics of XTX301, proteolytically cleaved XTX301 (XTX301 + MMP), and rhIL12 to IL12Rβ2. IL12Rβ2 was immobilized to a sensor chip. Then XTX301, proteolytically cleaved XTX301, and rhIL12 were flowed over at concentrations ranging from 3 nmol/L to 400 nmol/L. The concentrations of each analyte decrease from top to bottom within each panel. XTX301 activity was measured in a reporter cell line and primary human cells. B, IL12 HEK-Blue reporter gene cells were incubated with either rhIL12, unmasked control, or XTX301 at varying doses, and the reporter activity was measured. The data represents one of three independent experiments, data points represent the mean of 2 replicate wells and the error bars represent SD. C, Pre-activated primary human PBMCs were incubated with either rhIL12, unmasked control or XTX301 at varying doses, and STAT4 phosphorylation was assessed in CD8 + T cells via flow cytometry. The data points represent the mean of 2 replicate wells and the error bars represent SD. The data represents one of two independent experiments, each conducted with 2 different PBMC donors.

Article Snippet: Murine splenocytes were isolated via mechanical dissociation and pre-activated with PMA/Ionomycin (BioLegend, catalog #423301) for 2 days at 37°C before incubating with varying doses of rm IL12 (R&D Systems, catalog no. 419-ML/CF), murine surrogate mXTX301 or proteolytically activated mXTX301 for 24 hours.

Techniques: Activity Assay, In Vitro, Binding Assay, Incubation, Control, Phospho-proteomics, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: USP28 protects development of inflammation in mouse intestine by regulating STAT5 phosphorylation and IL22 production in T lymphocytes

doi: 10.3389/fimmu.2024.1401949

Figure Lengend Snippet: Luminex – plasma.

Article Snippet: CD8+ T cells were cultured with anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) in the presence of recombinant mouse IL12 (20 ng/ml), and mouse anti-IL4 (500-P54, 1 μg/ml, both from PeproTech).

Techniques: Luminex, Clinical Proteomics

Journal: Nature Communications

Article Title: FOXP3 + regulatory T cell perturbation mediated by the IFNγ-STAT1-IFITM3 feedback loop is essential for anti-tumor immunity

doi: 10.1038/s41467-023-44391-9

Figure Lengend Snippet: a Immunoblot analysis of IFITM3, STAT1, and FOXP3 in Treg cells after IL12 (50 ng/ml), IL27 (50 ng/ml), and IFNγ (50 ng/ml) treatment for 24 h. b Flow cytometric analysis of IFITM3 in WT and SKO Treg cells after IL12, IL27, and IFNγ treatment for 24 h ( n = 3). c RT–qPCR analysis of IFITM3 mRNA level in WT and SKO Treg cells after IL12, IL27, and IFNγ treatment for 24 h ( n = 3). d ChIP-qPCR analysis of −203 to −70bp and −23 to +109 bp of exon1 of Ifitm3 that is regulated by STAT1 in WT Treg cells. Data in d are representative of 3 independent experiments. e Tumor growth in WT, cKO, and IFNγ +/- cKO mice injected s.c. with MC38 murine colon cancer cells (WT: n = 7; cKO: n = 7; IFNγ +/- cKO: n = 7). f Tumor weight of WT, cKO, and IFNγ +/- cKO mice injected s.c. with MC38 murine colon cancer cells ( n = 4 per group). g Percentage of Treg cells in tumor of WT, cKO, and IFNγ +/- cKO mice injected s.c. with MC38 murine colon cancer cells (day 23, n = 4 per group). h , i Frequency of IL10 ( h ) and IFNγ ( i ) producing Treg cells in tumor of WT and cKO mice injected s.c. with MC38 murine colon cancer cells (day 23, n = 4 per group). j The quantification of the MFI of Ki67 in Treg cells in tumors of WT, cKO, and IFNγ +/- cKO mice injected s.c. with MC38 murine colon cancer cells (day 23, n = 4 per group). k , l Cytokine secretion of CD8 + ( k ) and CD4 + T cells ( l ) in tumor of WT, cKO, and IFNγ +/- cKO mice injected s.c. with MC38 murine colon cancer cells (day 23, n = 4 per group). The data f – l are presented as representative plots and as summary graphs ( n = 4 per group). Data are represented as the mean ± SD. Source data are provided as a Source Data file.

Article Snippet: The cytokines used in Treg in vitro experiments were Recombinant Mouse IL27 (577402, Biolegend, 50 ng/ml), Recombinant Mouse IL12 Protein (419-ML-010, R&D, 50 ng/ml), Recombinant Mouse IFN-gamma (485-MI-100/CF, R&D, 50 ng/ml)

Techniques: Western Blot, Quantitative RT-PCR, ChIP-qPCR, Injection