rae 1 delta recombinant mouse rae 1delta fc  (R&D Systems)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Toxicity induced by a bispecific T cell redirecting protein is mediated by both T cells and myeloid cells in immunocompetent mice 2

doi: 10.4049/jimmunol.1901401

Figure Lengend Snippet: (A) Diagram showing construction of treatment (mNKG2D-2C11) and control (TZ47-2C11) bsTCEs. (B-D) MC38 (1x106) cells were injected s.c. into WT B6 mice. Treatment was initiated when tumors reached approximately 40mm2. (B) Mice received a total of four 10μg i.v injections of bsTCE every other day. (Left) Health scoring was blinded and evaluated 3h and 24h after each treatment. (Right) Weight, normalized to day 0, at the indicated time points. Data pooled from 7 experiments (n=30) (C) Mice received a total of four i.v. injections of either 2, 5 or 10μg bsTCE every other day. Health scoring was blinded and evaluated every hour for the first 8h and then 24h after each treatment. Data pooled from 2 experiments (n=8). (D) Tumor or non-tumor bearing WT B6 mice received a total of four 10μg i.v injections of bsTCE every other day. Health scoring was blinded and evaluated 3h and 24h after each treatment. Data are pooled from two experiment (n=8). (E) Representative images of Rae1 positive staining in the liver of B6 mice or contiguous tissue sections blocked with recombinant Rae1. Slides counterstained with methylene blue. Health scores in B and D are shown +/− SD, weight and C are shown +/− SEM. Statistical significance determined by repeated measures two-way ANOVA with Bonferroni’s multiple comparisons test (B, C) or with Tukey multiple comparisons test (D). * p<0.05, *** p<0.001, NS = not statistically significant.

Article Snippet: Contiguous sections from each sample were incubated with primary antibodies pre-incubated for 4 hours with recombinant mouse Rae1 Fc chimeric protein (R&D Systems, 1998-RA) at a 1:10 molar ratio as specificity controls.

Techniques: Injection, Staining, Recombinant

Journal: Molecular Therapy

Article Title: T Cells Engineered With Chimeric Antigen Receptors Targeting NKG2D Ligands Display Lethal Toxicity in Mice

doi: 10.1038/mt.2015.119

Figure Lengend Snippet: In vitro functional profiles of NKG2D-CAR-T cells. (a,b) BALB/c or (a,c) C57BL/6 T cells were engineered with the indicated CARs. (a) NKG2D-CAR T cells were stimulated with plate-bound recombinant Rae1β-Fc fusion protein and stained for production of IFNγ and TNFα, and analyzed by flow cytometry. Data is expressed as mean frequency ± SEM normalized to background levels from three independent experiments. In vitro CAR-T cell-mediated cytotoxicity was assayed using 4T1.2 tumor cells using a 6-hour AlamarBlue assay at the indicated effector: target ratios of BALB/c (b) or C57BL/6 (c) T cells. Mean frequency of viable tumor cells ± SEM from 3–4 independent experiments of triplicate wells is presented.

Article Snippet: T cells were stimulated in round-bottom 96-well plates coated with 2,000 ng/ml recombinant mouse Rae1β-Fc chimera (R&D Systems, Minneapolis, MN) or 1,000 ng/ml recombinant human HER2-Fc chimera (R&D Systems).

Techniques: In Vitro, Functional Assay, Recombinant, Staining, Flow Cytometry, Alamar Blue Assay