Anti ctla4 antibody clone 9h10

In preliminary experiments, all PKF mice were observed to have more than 80% of the pancreas occupied by tumor at 42 days of age, so the timing was chosen for treatment initiation. 42-day-old PKF mice were treated with a single intratumoral injection with G47Δ (1 × 10⁷ pfu), intraperitoneal injections with anti-PD-L1 antibody (clone 10F.9G2, BioXCell, Lebanon, NH, USA, 200 μg/body), and anti-CTLA4 antibody (clone 9H10, BioXCell, 250 μg/body) every 4–5 days starting on day 42. For G47Δ treatment, mice were anesthetized, and the abdominal wall was incised 1.5 cm to expose the pancreatic tumor. G47Δ was injected with a 100-μL Hamilton syringe and a 30G needle at a depth of 5 mm. The abdominal wall was closed with a 5-0 nylon surgical suture.
For the combination therapy of FAKi and ICIs, 42-day-old PKF mice started daily treatment of oral administration with FAKi (50 mg/kg) and received intraperitoneal injections with anti-PD-L1 (200 μg/body) and anti-CTLA4 (250 μg/body) antibodies every 4–5 days since day 42. For the triple combination therapy, PKF mice were treated with FAKi, anti-PD-L1, and anti-CTLA4 in the same manner as above, and a single intratumoral injection with G47Δ (1 × 10⁷ pfu) on day 52. The rat IgG2b antibody (clone LTF-2, BioXCell) and Syrian Hamster IgG antibody (polyclonal, BioXCell) were used as isotype controls.

Tumor nodules reached 50 mm³ on day 3. Mice were randomly divided into four groups as follows: vehicle, anti-CTLA-4 antibody alone (a-CTLA-4); MMPI alone, and combination therapy (a-CTLA-4 plus MMPI). Mice were treated with 100 µg anti-CTLA-4 antibody (clone, 9H10; BioXCell, West Lebanon, NH, USA) by intraperitoneal (i.p.) injection once every two day and/or with 0.1 MMPI (1 mg/kg) by subcutaneous injection once every two days. The MMPI potassium ferricyanide {K₃[Fe(CN)₆ (10 (link),11 (link)), was kindly provided by Professor Xuexun Fang (Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, China). Mice in the vehicle group were treated with an equal volume of normal saline and PBS. The dosages used in the combined treatment group were equal to those used in the monotherapy groups. The longest (a) and shortest (b) diameters of the tumor were measured with calipers every 3 days, and tumor volume was estimated by the formula V = ab²/2. On days 7, 21 and 35, tumors were measured using an Ultrasound Biomicroscopy InviVue instrument (PanoView β1500; Taiwan).