Female 6 week old BALB/c or C57BL/6 mice were inoculated s.c. in the flank on day ~−7 with 5×105 CT26 colon carcinoma or 1×106 B16F10 melanoma cells, respectively. Intra-tumoral (i.t.) injections of mouse CD80-Fc (20μg/50μl/mouse, R&D Systems), rat IgG2a clone 2A3 (20μg/50μl/mouse, BioXCell), or CpG ODN 1555 (100μg/50μl/mouse) were started on day 0 when tumors were approximately 5mm in diameter and administered twice a week for three weeks (7 (link)). For some experiments mice were inoculated on ~day −7 with B16F10 cells on both flanks and tumors on only one flank were treated. Intraperitoneal (i.p.) injections of mouse CD80-Fc (200ug/mouse, R&D Systems), rat IgG2a clone 2A3 (200ug/mouse BioXCell), or anti-mouse PDL1 clone 10F.9G2 (200ug/mouse BioXCell) were administered on days 3, 6, 9, and 22 following tumor inoculation. Mice were sacrificed when they became moribund and their tumors were cryopreserved and analyzed by immunohistochemistry. Tumors were measured every 2–3 days in two perpendicular diameters. Tumor volume = 4/3πr3 where r = (diameter 1 + diameter 2)/4.
Protocol detail
Tumor-directed immune modulation therapy
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Anti pdl1
C57bl mice
Cells
Clone
Colon carcinoma
Cpg odn 1555
Female
Igg2a
Immunohistochemistry
Melanoma
Mice
Tumor inoculation
Tumors
Corresponding Organization : University of Maryland, Baltimore County
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Variable analysis
independent variables
- Intra-tumoral (i.t.) injections of mouse CD80-Fc (20μg/50μl/mouse, R&D Systems)
- Intra-tumoral (i.t.) injections of rat IgG2a clone 2A3 (20μg/50μl/mouse, BioXCell)
- Intra-tumoral (i.t.) injections of CpG ODN 1555 (100μg/50μl/mouse)
- Intraperitoneal (i.p.) injections of mouse CD80-Fc (200ug/mouse, R&D Systems)
- Intraperitoneal (i.p.) injections of rat IgG2a clone 2A3 (200ug/mouse BioXCell)
- Intraperitoneal (i.p.) injections of anti-mouse PDL1 clone 10F.9G2 (200ug/mouse BioXCell)
- Tumor volume (calculated as 4/3πr3 where r = (diameter 1 + diameter 2)/4)
- Tumor cryopreservation and analysis by immunohistochemistry
- Female 6 week old BALB/c or C57BL/6 mice
- Subcutaneous (s.c.) inoculation of 5×105 CT26 colon carcinoma or 1×106 B16F10 melanoma cells on day ~−7
- Tumors were approximately 5mm in diameter on day 0 when treatments started
- Treatments were administered twice a week for three weeks
- Positive control: Intra-tumoral (i.t.) injections of rat IgG2a clone 2A3 (20μg/50μl/mouse, BioXCell)
- Negative control: Not explicitly mentioned
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