For mammary tumor models, 5 × 104 EMT6 or 1 × 105 E0771 cells were orthotopically injected into the fourth right mammary fat pad of female BALB/c (EMT6) or C57BL/6J (E0771) mice. Following the establishment of tumors (∼100–200 mm3) mice were stratified prior to therapy administration. The mice were treated via intraperitoneal (i.p.) injection with isotype IgG2a control (Bio X Cell, clone BE0089), anti-NKG2A (Bio X Cell, clone 20D5), or anti–PD-L1 (Genentech, clone 6E11) dosing at 200 μg on day 7 and 100 μg on days 14, 21, 28, and 35. Mice used for each treatment arm are defined in respective figure legends. For tumor growth analysis, tumors were measured 3 times weekly with calipers, and volume was calculated in mm3 using the formula (length × width × width/2). Mice were humanely euthanized at defined end points or when the tumor volume reached 2 cm3 or tumor ulceration. For the pINDUCER tumor model, mice were primed with doxycycline (2 mg/mL in 5% sucrose, ad libitum) or 5% sucrose (control) for 2 days before injection. Doxycycline was continually administered in drinking water before sacrifice.
Clone 6e11
Manufactured by Genentech
Clone 6E11 is a laboratory equipment product. It is designed for the isolation and purification of specific proteins or macromolecules from biological samples. The core function of Clone 6E11 is to facilitate the cloning and expression of target DNA sequences.
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Lab products found in correlation
4 protocols using clone 6e11
1
Immunotherapy Testing in Mammary Tumor Models
2
Intracranial Tumor Implantation and Anti-PD-L1 Treatment in Mice
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Wild-type C57BL/6 mice were purchased from Janvier Labs (Le Genest-Saint-Isle, France). RAG−/− mice were bred at the University of Zurich in pathogen-free facilities. The mice underwent surgery at age 6-12 weeks. Stereotactic intracranial tumor cell implantation was made into the right striatum; 80,000 cells were implanted for CT-2A, 100,000 cells were implanted for GL-261, and 20,000 cells were implanted for SMA-560. Mice were regularly monitored and euthanized as indicated or when developing neurological symptoms grade 2; weight loss above 15% compared with weight at day of tumor cell implantation, moderate signs of pain, slight paralysis of left leg or no/decreased activity. Where indicated, mice received anti-PD-L1 (Genentech Clone 6E11) twice weekly, 5 mg/kg intra-peritoneally. Treatments were started 5 days after tumor cell implantation. Anti-PD-L1 treatment was given for 3 consecutive weeks. All animal study procedures were approved by Swiss Cantonal Veterinary office under animal licenses ZH098/2018 and ZH109/2020.
3
Combination Immunotherapy with Paclitaxel, Sunitinib, and Anti-PD-L1 Antibodies
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Paclitaxel (Accord Healthcare Inc., DIN: 02391465) was administered at 30 mg/kg, every 2 weeks, intraperitoneally (IP); phosphate-buffered saline (PBS) served as its control. Sunitinib malate (LC Laboratories; administered at 60 mg/kg daily) and its vehicle, formulated as previously described,42 (link) were administered by oral gavage (PO). The therapeutic anti-VEGF agent—a mouse IgG2a antibody against mouse/human VEGF (Genentech, clone B20-4.1.1)—was administered at 5 mg/kg, 2×/week, IP. The two therapeutic antibodies to mouse PD-L1 (also known as CD274 or B7-H1) were a mouse IgG1 raised in PD-L1 knockout mice with a C57BL/6 genetic background (Genentech, clone 6E11; administered at 5 mg/kg, 2×/week, IP) and a rat IgG2b monoclonal antibody (BXCell InVivoMAb #BE0101, clone 10F.9G2; administered at 100 μg/dose, 2×/week, IP)—both antibodies block PD-L1:PD-1 and PD-L1:B7.1 (CD80) interactions. The corresponding InVivoMAb controls from BioXCell were: mouse IgG2a isotype control (#BE0085, clone C1.18.4, unknown specificity), mouse IgG1 isotype control (#BE0083, clone MOPC-21, unknown specificity), and rat IgG2b isotype control (#BE0090, clone LTF-2, anti-keyhole limpet haemocyanin).
4
Mammary Tumor Model and Immunotherapy
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For mammary tumor models, 5 × 104 EMT6 were orthotopically injected into the fourth left mammary fat pad of female BALB/c mice. Cells were tested for Mycoplasma contamination prior to each experiment using the e-Myco Mycoplasma PCR Detection Kit (LiliF Diagnostics). Following the establishment of tumors (∼100–200 mm3), mice were stratified prior to therapy administration. Mice were treated via intraperitoneal injection with isotype IgG1 control (BioXcell, clone BE0083) or anti-PD-L1 (Genentech, clone 6E11) dosing at 200 µg for the first treatment and 100 µg for two subsequent treatments at 1-week intervals. For tumor growth analysis, tumors were measured two to three times weekly with calipers, and volume was calculated in mm3 using the formula (length × width × width/2). Mice were humanely euthanized at defined endpoints or when the tumor volume reached 2,000 mm3 or tumor ulceration.
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