human il 7 elisa kit  (Elabscience Biotechnology)

Journal: Life Medicine

Article Title: Integration of a novel anti-PD-1 antibody with chimeric antigen receptor-T engineered to express interleukin-7 enhances targeting efficacy against lung cancer

doi: 10.1093/lifemedi/lnaf035

Figure Lengend Snippet: Engineering IL-7 in CAR-T cells enhances their anti-tumor activity .(A) Measurement of IL-7 secretion in culture supernatant by ELISA. Data are presented as the mean ± SD, n = 3, **** P < 0.0001. (B) Assessment of proliferation capacity of IL-7-CAR-T cells by cell count. NT, CAR-T, and IL-7-CAR-T cells were cultured with an initial cell count of 2 × 10 5 , and cell numbers were counted every other day. Data shown are the mean ± SD, n = 3; * P < 0.05, Student’s t -test. (C) The expression of CD45RA and CD62L on CAR-T cells and IL-7-CAR-T cells were analyzed by FACS. (D) Statistical analysis of memory T cell phenotypes. Data are shown as the mean ± SD, n = 3, * P < 0.05, Student’s t -test. (E) Evaluation of specific cytotoxicity of IL-7-CAR-T cells against lung cancer cell lines (HCC827, H23) using luciferase-based assays at various effector-to-target (E: T) ratios (10:1, 3:1, and 1: 1), with K562 cells as a negative control. Data are presented as the mean ± SD, n = 3, * P < 0.05, ** P < 0.01, Student’s t -test. (F) The experimental timeline for the treatment of cell line-derived xenograft (CDX) tumor model. 4 × 10 5 HCC827/Luc cells were subcutaneously injected into NPG mice. After 4 days, tumor-bearing NPG mice were treated intravenously with PBS, 1 × 10 7 NT, CAR-T, and IL-7-CAR-T cells. (G) Peripheral blood T-cell counts were quantified by flow cytometry at 10 days post-adoptive transfer. Data are shown as the mean ± SD, n = 3; ** P < 0.01, Student’s t -test. (H) Evaluation of the anti-tumor activity of IL-7-CAR-T cells by in vivo imaging in mice bearing HCC827 xenografts. Tumor growth was monitored over 28 days. (I) Bioluminescence signals from each mouse, as in (H), were recorded every 7 days. (J) The survival of tumor-bearing mice treated with PBS, NT, CAR-T cells and IL-7-CAR-T cells. PBS, CAR-T cells and IL-7-CAR-T cells group n = 5, NT group n = 3, * P < 0.05, ** P < 0.01, log-rank test. (K) Tumor-infiltrating T cell populations were quantified by flow cytometry at 10 days post-adoptive transfer. Data are shown as the mean ± SD, n = 3; **** P < 0.0001, Student’s t -test.

Article Snippet: IL-7 cytokine secretion in the supernatants of NT, CAR-T, and IL-7-CAR-T cells was quantified using ELISA Kits (Multi-Science, EK107-96).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Cell Counting, Cell Culture, Expressing, Luciferase, Negative Control, Derivative Assay, Injection, Flow Cytometry, Adoptive Transfer Assay, In Vivo Imaging

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: Engineering CAR T cells to secrete and efficiently utilize IL-7 cytokine (A). Illustrations of the IL-7 cytokine construct containing mOrange fluorescent protein for transgene detection by flow cytometry (top) and detection of both the PSCA CAR and IL-7 transgene expression in C.P7 T cells from a representative donor after serial transduction (bottom). (B) Summary data indicating PSCA CAR and IL-7 double-transduced T cells compared to NT cells (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Production of IL-7 by C.P7 cells after stimulation with irradiated K562 cells engineered to express PSCA, measuring using ELISA (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (D) Diagram illustrating the IL-7Rα construct containing GFP for transgene detection (top) and flow cytometry data for a representative donor demonstrating the expression of both the MUC1 CAR and the IL-7Rα transgenes in C.M7R T cells. (E) Summary data comparing IL-7Rα detection by flow cytometry in NT, C.M, and C.M7R cells (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (F) Quantification of C.M and C.M7R T cells using trypan blue exclusion during culture with irradiated CAPAN1 tumor cells in presence or absence of recombinant IL-7 cytokine (t-tests on day 8, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Article Snippet: Human Duoset ELISA kits from R&D Systems (Minneapolis, MN) were used to measure IL-7, IFN-γ, and TNF-α cytokines produced by T cells.

Techniques: Construct, Flow Cytometry, Expressing, Transduction, Irradiation, Enzyme-linked Immunosorbent Assay, Recombinant