Treatment diagrams are provided for each specific experiment. APD-1 treatments consisted of systemically (intraperitoneally) delivered antibody dosed as 0.1 mg (clone 10F.9G2, BE0101, BioXCell, Lebanon, New Hampshire, USA) diluted in PBS. Virotherapy treatments consisted of 1 x 108 viral particles (including equal amounts of Ad5-CMV-mIL2 and Ad5-CMV-mTNFa viruses, non-replicative in mice but historically used as model for the Ad5 man-based therapies which are replication competent in that host). The expression of TNFa and IL-2 after the use of the viruses in B16.OVA model has been studied before (23 (link)). In those experiments including intratumoral virus treatments, control groups not receiving viruses were injected intratumorally with an equivalent amount of PBS.
Clone 10f 9g2
Manufactured by BioXCell
Sourced in United States
Clone 10F.9G2 is a laboratory equipment product. It is a research tool designed for cellular and molecular biology applications. The core function of Clone 10F.9G2 is to facilitate cell culture and manipulation experiments.
Automatically generated - may contain errors
Lab products found in correlation
Clone ltf 2, by BioXCell (4 mentions)
Clone 2a3, by BioXCell (2 mentions)
Clone 9d9, by BioXCell (2 mentions)
Clone rmp1 14, by BioXCell (2 mentions)
Be0101, by BioXCell (1 mentions)
Matrigel, by BD (1 mentions)
Anti pd l1, by BioXCell (1 mentions)
Anti ccl5, by R&D Systems (1 mentions)
Panc02, by American Type Culture Collection (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Clone mp5 20f3, by BioXCell (1 mentions)
Ltf 2, by BioXCell (1 mentions)
Naive cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Anti pd l1 mab, by BioXCell (1 mentions)
Be0061, by BioXCell (1 mentions)
Be0090, by BioXCell (1 mentions)
Azd1775, by Selleck Chemicals (1 mentions)
Recombinant murine il 2, by Thermo Fisher Scientific (1 mentions)
Clone xt3, by BioXCell (1 mentions)
Nod shiltj, by Jackson ImmunoResearch (1 mentions)
25 protocols using clone 10f 9g2
1
Systemic and Intratumoral Immunotherapy Protocols
2
Immunotherapy Efficacy in Murine Tumor Models
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Mice were housed in specific pathogen-free (SPF) conditions, and animal protocols were authorized by the local ethics committee. For both the subcutaneous and orthotopic tumor models, mice were treated with isotype IgG (200 μg/mouse, q3d, clone LTF-2, BioXcell), anti-PD-L1 (200 μg/mouse, q3d, clone 10F.9G2, BioXcell) or combined with anti-CCL5 (20 μg/mouse, R&D, Minneapolis, MN) intraperitoneally q3d. The anti-CD8 antibody (No. 2.43, BioXCell, UK) for eradicating CD8+ T cells was i.p. injected at a dose of 250 µg on days 6, 9, 15, and 21. Tumor volumes were measured every three days with a caliper. For the orthotopic model, C57BL/6 mice were injected with 1 × 106 Pan02-pLV-control-luc or Pan02-pLV-FOXP3-luc cells in Matrigel (BD Biosciences) into the pancreatic tail. Tumor growth was analyzed by bioluminescent imaging. The survival time of each mouse was recorded. Further information is provided in the Supplementary Materials and Methods .
3
Iminothiolation of Anti-PD-L1 Antibody
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Iminothiolation of αPD‐L1 was conducted according to a previous study[20 ] and the manufacturer's protocol. One milligram of anti‐mouse PD‐L1 antibody (BioXcell Inc., clone 10F.9G2) and 40 µg of 2‐iminothiolane were diluted to a final volume of 0.5 mL in sodium phosphate buffer (pH 8). The mixture was incubated at 4 °C for 2 h in a rotating chamber, followed by repeated centrifugation at 4000 rpm and 4 °C for 30 min using an Ultra‐4 centrifugal filter unit (Amicon, Billerica, MA) to remove excess 2‐iminothiolane.
4
Murine Pancreatic Cancer Cell Lines
5
Therapeutic Vaccination and Immune Checkpoint Blockade
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Vaccinated mice were challenged 14 days after their last immunization by subcutaneous injection of 1 × 105 B16vIII cells into the right flank of recipient mice. Tumor growth was monitored every other day, and the tumor volume was calculated as follows: volume = length × width2 × 0.52. Animals were euthanized when the masses reached 1.5 cm in diameter, when tumor volume reached 2000 mm3, or when ulceration occurred. For the therapeutic tumor model, mice were inoculated with 1 × 105 B16vIII cells in 200 μl of 1× PBS into the right flank. Vaccinations were given 1 day after tumor inoculation, and booster immunizations were given 14 days after primary vaccination. For combinatorial immunotherapy, anti-mouse PD-L1 (100 μg per mouse, clone 10F.9G2, BioXCell) or IgG2b isotype control (100 μg per mouse, clone LTF-2, BioXCell) was administered intraperitoneally on indicated days, and anti-mouse CD47 (50 μg per mouse, clone MIAP301, BioXCell) or IgG2a isotype control (50 μg per mouse, clone 2A3, BioXCell) was administered intratumorally on indicated days. Tumor growth was determined as described above.
6
Activation of 2D2 CD4+ T Cells by DCs
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Naive CD4+ T cells from spleens of 2D2 mice were isolated using magnetic beads (Naive CD4+ T cell isolation kit, Miltenyi Biotec, CA, USA). 2 × 105 naive CD4+ T cells were added to each well of the cell culture plate containing moDCs, cDC1, or BMDCs (ratio of 1 DC: 10 T cells) and plates were incubated at 37°C in the presence of MOG35−55 peptide (20 μg/mL) and anti-PD-L1 MAb (1 μg/mL; clone 10F.9G2, BioXCell). Cells were collected after 72 h and analyzed by flow cytometry, while cytokine concentrations in culture supernatants were measured by ELISA.
7
Anti-PD-L1 Therapy in MMTV-PyMT Tumor Model
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Tumor cells from MMTV-PyMT;Ccr2+/+ or MMTV-PyMT;Ccr2−/− mice were transplanted as described above. For anti–PD-L1 treatment, on days 15, 18, 21, and 24 after transplantation (after tumors had formed), mice received 200 µg of anti–PD-L1 antibody by intraperitoneal injection (Clone 10F.9G2; Bio X Cell) or control rat IgG2b antibody (Bio X Cell; Winograd et al., 2015 (link)). For CD8+ T cell depletion, mice were injected intraperitoneally with 200 µg of anti-CD8a antibody (Clone 2.43; BE0061; Bio X Cell) or control rat IgG2b antibody (Clone LTF-2; BE0090; Bio X Cell) every 3 d starting from day 0 and until day 30. Tumor size was measured biweekly, and mice were sacrificed at the IACUC-approved endpoint.
8
Targeting PD-L1 and CTLA-4 Pathways
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JQ1 (Sigma-Aldrich), an inhibitor of PD-L1 expression was used in culture as described (24 (link)). Blocking anti-mouse PD-L1, clone 10F.9G2 and anti-mouse CTLA-4 (CD152), clone 9H10 (BioXCell) were used as described (25 (link)).
9
Hepatocellular Carcinoma Induction and Anti-PD-L1 Immunotherapy
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FVB mice were acquired from the Institute of Zoology, Chinese Academy of Sciences (Beijing, China), and were housed under specific pathogen-free conditions, with free access to food and water. All animal experiments were conducted in accordance with the guidelines approved by the Animal Ethics Committee of our institution. After a standardized acclimatization period, hepatocellular carcinoma (HCC) induction commenced. To induce HCC, mice were subjected to a carefully calibrated dose regimen of diethylnitrosamine (DEN, Sigma-Aldrich, St. Louis, MO, USA), a potent hepatocarcinogen. DEN was administered through intraperitoneal injection starting with a dose of 20 mg/kg body weight when the mice were 15 days old, followed by a dose of 30 mg/kg in the third week, and then 50 mg/kg for the last 6 weeks. For the administration of immunotherapy, the anti-PD-L1 monoclonal antibody Clone 10F.9G2, Bio X Cell, West Lebanon, NH, USA was selected, with intraperitoneal injections of 100 µg per mouse administered twice a week.
10
Synergistic Cancer Immunotherapy Protocol
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CT26 cells (5 × 105) were subcutaneously injected into the lower abdomen of male mice to establish the CT26 tumor model. Ten days after inoculation, CT26 tumor-bearing mice were randomly divided into 5 groups: Vehicle, Vehicle+RT, aAGd-NWs+RT, aAGd-NWs+RT+αCD8a and aAGd-NWs+RT + αPD-L1 ([Gd] = 15.7 mg kg−1, ([ara-AMP] = 34.7 mg kg−1), [αCD8a] = 10 mg kg−1 and [αPD-L1] = 10 mg kg−1). Treatments were performed on day 0 and day 6, respectively, and followed by radiotherapy (5 Gy × 2) 6 h later. 6 h post X-ray irradiation, mice were intraperitoneally injected with anti-PD-L1 antibody (10 mg kg−1 × 4 with fractions delivered 3 days apart, Clone: 10 F.9G2, BioXcell, America) and anti-CD8a antibody (10 mg kg−1 × 4 with fractions delivered 3 days apart, Clone: 53-6.7, BioLegend, America), respectively.
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