TSA Mouse Breast Carcinoma Tumor Growth Study
Objective: Test whether fractionated radiotherapy regimens synergize with anti-CTLA-4 antibody to induce tumor growth delay and abscopal effects in syngeneic mouse carcinoma models
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Equipment1
Not specified • Not mentioned • Not mentioned • Not mentioned
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Protocol Steps
Tumor cell injection
TSA mouse breast carcinoma cells were injected subcutaneously into syngeneic mice at two separate sites
Note: One site designated as primary (to be irradiated), one as secondary (outside radiation field)
View evidence from paper
“TSA mouse breast carcinoma cells were injected s.c. into syngeneic mice at two separate sites, defined as a primary site that was irradiated, and a secondary site outside the radiotherapy field”
Tumor palpation and randomization
When both tumors became palpable, mice were randomly assigned to 8 treatment groups
Note: Randomization occurred when both primary and secondary tumors were palpable
View evidence from paper
“When both tumors were palpable mice were randomly assigned to 8 groups”
Treatment group assignment
Mice were assigned to 8 groups: control (no radiotherapy), three radiotherapy-only regimens, anti-CTLA-4 antibody alone, or combinations of radiotherapy with anti-CTLA-4 antibody
Note: 8 total groups tested different treatment combinations
View evidence from paper
“receiving no radiotherapy or 3 distinct regimens of radiotherapy (20 Gy × 1, 8 Gy × 3 or 6 Gy × 5 fractions in consecutive days) in combination or not with 9H10 mAb against CTLA-4”
Radiotherapy administration - single dose regimen
Primary tumors received single dose radiotherapy of 20 Gy in one fraction
Note: Single dose regimen tested as comparison to fractionated approaches
View evidence from paper
“20 Gy × 1”
Radiotherapy administration - fractionated regimen 1
Primary tumors received fractionated radiotherapy of 8 Gy per fraction for 3 fractions on consecutive days
Note: Fractionated regimen expected to synergize with anti-CTLA-4
View evidence from paper
“8 Gy × 3 or 6 Gy × 5 fractions in consecutive days”
Radiotherapy administration - fractionated regimen 2
Primary tumors received fractionated radiotherapy of 6 Gy per fraction for 5 fractions on consecutive days
Note: Fractionated regimen expected to synergize with anti-CTLA-4
View evidence from paper
“6 Gy × 5 fractions in consecutive days”
Anti-CTLA-4 antibody administration
9H10 monoclonal antibody against CTLA-4 administered to designated treatment groups
Note: Administered alone or in combination with radiotherapy regimens
View evidence from paper
“9H10 mAb against CTLA-4”
Tumor monitoring
Mice were followed for tumor growth and regression at both primary and secondary sites
Note: Primary tumors were in radiation field; secondary tumors were outside radiation field to assess abscopal effects
View evidence from paper
“Mice were followed for tumors growth/regression”
Immune analysis
Frequency of CD8+ T cells showing tumor-specific IFN-γ production was measured and correlated with secondary tumor inhibition
Note: Immune response measured to understand mechanism of abscopal effect
View evidence from paper
“Frequency of CD8+ T cells showing tumor-specific IFN-γ production was proportional to the inhibition of the secondary tumor”