Anti pd1 antibody clone rpm1 14

UAMC-1110 was generously provided by Dr. Pieter Van der Veken (University of Antwerp). Unless otherwise specified, UAMC-1110 was administered in high molecular weight PEG at a concentration of 20mg/kg by oral gavage twice per day. Fluorescently-conjugated antibodies CD3-e450, CD8-PerCP, CD4-FITC, CD4-e450, CD4-PerCP, CD25-APC, IFNγ-APC, IL-2-PE, TNFa-PE-Cy7, CD11b-PE-Cy7, Ly6G-FITC, Ly6C-PerCP-Cy5.5, Gr1-PE-Cy7, and MHCII-EF450, PDGFRβ-APC, CD31-PE, CD45-BV510, and EpCam-BV605 were purchased from Thermo Fisher Scientific (Lafayette, CO) or BD Biosciences (San Jose, CA). CD8-PE-TxRD was purchased from Invitrogen (Carlsbad, CA). SIY, SIINFEKL, and β-galactosidase peptides were obtained from Integrated DNA Technologies (Coralville, Iowa). β-galactosidase (βgal) for the ICPMYARV peptide and SIINFEKL peptide tetramers were obtained from the NIH Tetramer core facility (Atlanta, GA). Antibodies to CD31 (Thermo Fisher Scientific), αSMA (Sigma-Aldrich, St Louis, MO), PDGFRα (Thermo Fisher Scientific), Vimentin (Cell Signaling Technology, MA), Ki67 (Abcam), Cleaved Caspase 3 (Cell Signaling Technology) were used for immunofluorescent staining and hypoxyprobe Omni Kit (Burlington, MA) was used for hypoxia staining. Anti-PD1 antibody clone RPM1-14 (BioXcell, West Lebanon, NH) was used where indicated for treatment studies.

Eight to twelve weeks old female hOX40tg mice were injected subcutaneously (sc) on the right hind flank on day 0 with MB49 cells (0.25 × 10⁶ cells) or MC38 cells (0.9 × 10⁶ cells). Vehicle (PBS) or antibodies, including ATOR-1015, monotargeting anti-CTLA-4 and anti-OX40 antibodies, IgG1 control, anti-PD-1 antibody (clone RPM1–14, BioXcell) or surrogate anti-CTLA-4 antibodies (clones 9D9 and 9H10, both BioXcell), were administered intraperitoneally (ip) at indicated time-points. For anti-tumor efficacy studies, tumor volume was measured three times weekly with a caliper and calculated as: (width/2 × length/2 × height/2) × 4π/3. Animals were euthanized when the ethical human endpoints were reached, including tumor volume exceeding 2 cm³, tumor ulceration or affected health.