Male wild-type and Rag1−/− C57BL/6J mice were purchased from Jackson laboratories. All mouse procedures were approved by the Institutional Animal Care and Use Committee of Yale University and conducted in accordance with the guidelines. Animals are housed in Yale Animal Resources Facilities with 12h:12h light:dark cycle, at ambient temperature of 68–79°F and humidity of 30–70%. Five to eight-week old mice were age-matched for tumor inoculation. Group sizes were selected based upon prior knowledge. Mice were age, gender and genetic background-matched and randomized appropriately, but blinding was not done. For YUMMER1.7 cell, 0.5×106 cells in 100μl PBS were injected. For YUMM1.7 cell, 100,000 cells were injected. For CD8+ T cell depletion experiments, 200 μg of anti-mouse CD8α (2.43, Bio X Cell) or Rat IgG (IRTSDIGGGFLY100MG, Innovative Research) were intraperitoneally injected 2 days before tumor injection, and this treatment continued twice a week until tumor size reached the end point (1,000 mm3). For immunotherapy treatment, anti-PD-1 antibody (RMP1–14) were purchased from Bio X cell. Mice were treated with 100 μg anti-PD-1 or Rat IgG twice per week for 3 weeks since day 7 after tumor injection for YUMM1.7. Tumor volumes were calculated using the equation: 0.5233*l*w*h. The humane endpoints were determined by animal discomfort level and tumor sizes.
Anti pd 1 antibody rmp1 14
Manufactured by BioXCell
Sourced in United States
The Anti-PD-1 antibody (RMP1–14) is a laboratory reagent used in research applications. It functions as a monoclonal antibody that binds to the PD-1 (Programmed Cell Death Protein 1) receptor on the surface of cells.
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3 protocols using anti pd 1 antibody rmp1 14
1
Murine Melanoma Model with Immunotherapy
2
Anti-PD-1 Therapy and Gefitinib in 4T1 Mouse Model
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All BALB/C mouse (6–8-week-old female; Jackson Laboratories) procedures were conducted under the guidelines approved by the Institutional Animal Care and Use Committee at MD Anderson Cancer Center. Mice were divided according to the mean value of tumour volume in each group. 4T1-Luc cells (5 × 104 cells in 50 μl medium mixed with 50 μl Matrixgel Basement Membrane Matrix (BD Biosciences, San Jose, CA, USA) were injected into the mammary fat fad. For treatment with antibody, 100 μg anti-PD-1 antibody (RMP1-14, Bio X Cell, West Lebanon, NH, USA) or rat IgG (Bio X Cell) as control was injected intraperitoneally on days 3, 6, 9 and 12 after 4T1 cell inoculation. For drug treatment, mice were treated with daily oral doses of 10 mg kg−1 gefitinib for 2 weeks (5 days per week). Tumour was measured weekly with a caliper, and tumour volume was calculated using the formula: π/6 × length × width2.
3
Murine Tumor Growth Assay with Immunotherapy
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Tumor cells were subcutaneously injected into mice and tumor growth was analyzed as described previously18 (link). Briefly, we intradermally injected 3 × 105 cells of B16F10, 1 × 106 of LLC, MB49, and 4T1, or 2 × 106 cells of MC38 cells into the back of the mice. After inoculation of the mice, the diameters of the tumors were measured with a caliper and the tumor volume was determined according to the following formula: tumor volume (mm3) = (length) × (width)2 × 0.5. Regarding survival analysis, the mice were euthanized when the tumor volume reached 4000 mm3, and the time for tumor volume to reach this endpoint was evaluated. ERB was kindly gifted by Eisai Co., Ltd and 10 µg of this reagent was intravenously administered to mice on day 2 after tumor inoculation. Anti-PD-1 antibody (RMP1–14) was purchased from Bio X Cell (Lebanon). The mice were given intraperitoneal injections of anti-PD-1 antibody (200 μg/mouse) every 4 days from day 4 to day 16. All the experiments were performed in accordance with the guidelines of the animal ethics committee of the University of Tsukuba Animal Research Center (permission number: #22–237).
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