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. Author manuscript; available in PMC: 2011 Jun 15.

Published in final edited form as: Int J Cancer. 2010 Jun 15;126(12):2893–2903. doi: 10.1002/ijc.24995

(a) Wild type C57BL/6 mice, treated with lapatinib (75mg/kg/d) or vehicle (PBS), were vaccinated with Ad-HER2-ki, Ad-LacZ, or vehicle (PBS). The magnitude of the immune response to HER2 was measured by IFNγ ELISPOT. The mean number of IFNγ producing cells per 500,000 splenocytes is represented +/− SD. (b) Mouse 4T1-HER2 (HER2+) and 4T1 (HER2−) were incubated with diluted HER2-vaccine induced antibodies or LacZ-vaccine induced antibodies, stained with PE-conjugated anti-mouse IgG and samples analyzed by flow cytometer. The histogram representing the mean fluorescent intensity (MFI) for each treatment is shown. (c) 4T1 or 4T1-HER2 cells were incubated mouse HER2-vaccine induced antibodies (from mice treated with or without lapatinib as in figure 1A) or LacZ-vaccine induced antibodies (1:50) along with rabbit serum as a source of complement. Supernatant from the cultures were taken for the CDC assay. The percentage of lysis was calculated by the following formula: lysis(%)=(experimental−target spontaneous)/(target maximum−target spontaneous)*100%. (d) NK cells derived from C57BL6 mice were cultured with the cell line SKBR3 and serum from mice treated with Lapatinib, Ad-LacZ, Ad-LacZ plus lapatinib, Ad-HER2-ki, Ad-HER2-ki plus lapatinib. Trastuzumab was used as a control for ADCC activity. The percentage lysis of the tumor cells (+/− std dev) was determined by Chromium release assay.

(a) Wild type C57BL/6 mice, treated with lapatinib (75mg/kg/d) or vehicle (PBS), were vaccinated with Ad-HER2-ki, Ad-LacZ, or vehicle (PBS). The magnitude of the immune response to HER2 was measured by IFNγ ELISPOT. The mean number of IFNγ producing cells per 500,000 splenocytes is represented +/− SD. (b) Mouse 4T1-HER2 (HER2+) and 4T1 (HER2−) were incubated with diluted HER2-vaccine induced antibodies or LacZ-vaccine induced antibodies, stained with PE-conjugated anti-mouse IgG and samples analyzed by flow cytometer. The histogram representing the mean fluorescent intensity (MFI) for each treatment is shown. (c) 4T1 or 4T1-HER2 cells were incubated mouse HER2-vaccine induced antibodies (from mice treated with or without lapatinib as in figure 1A) or LacZ-vaccine induced antibodies (1:50) along with rabbit serum as a source of complement. Supernatant from the cultures were taken for the CDC assay. The percentage of lysis was calculated by the following formula: lysis(%)=(experimental−target spontaneous)/(target maximum−target spontaneous)*100%. (d) NK cells derived from C57BL6 mice were cultured with the cell line SKBR3 and serum from mice treated with Lapatinib, Ad-LacZ, Ad-LacZ plus lapatinib, Ad-HER2-ki, Ad-HER2-ki plus lapatinib. Trastuzumab was used as a control for ADCC activity. The percentage lysis of the tumor cells (+/− std dev) was determined by Chromium release assay.