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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) HER2-specific T cell responses in vaccinated mice. Pooled splenocytes obtained from C57BL/6 mice (n=8 per treatment group) 14 days after injection of Ad-HER2-ki, Ad-LacZ, or saline, were tested in an interferon-gamma ELISPOT and the number of responding T cells (+/− std deviation) was determined. The number of responding cells per 500,000 splenocytes in response to the antigen stimulus (x-axis) is shown. (b) HER2-specific T cell responses in vaccinated mice. Serum obtained from the same mice as in A at 14 days post injection of Ad-HER2-ki or Ad-LacZ was mixed with the listed cell lines shown on the x-axis. Binding of anti-HER2 antibodies was detected by ELISA. Absorbance was measured at 660 nm. (c) Polyclonal anti-HER2 Vaccine Induced Antibodies (Ad-HER2-VIA) mediates CDC in vitro. Pooled serum (8 mice per group) from C57BL/6 mice immunized with Ad-HER2-ki or Ad-LacZ, or, trastuzumab (10 μg/ml), was mixed with HER2 overexpressing cell lines (BT474 and SKBR3) or a HER2 negative cell line (MDA-231) and then rabbit complement was added. The percentage lysis of the cells (+/− std dev) was determined by Chromium release assay. (d) Polyclonal anti-HER2 Vaccine Induced Antibodies (Ad-HER2-VIA) mediates ADCC in vitro. NK cells derived from C57BL/6 mice were cultured with the anti-HER2 vaccine induced antibodies, anti-LacZ vaccine induced antibodies, or trastuzumab, and the cell line 4TI-HER2. The percentage lysis of the cells (+/− std dev) was determined by Chromium release assay. (e) Anti-proliferative effect of HER2-vaccine induced antibodies on HER2+ SKBR3 proliferation. HER2+ SKBR3 cells were cultured with HER2-vaccine induced antibodies, LacZ-vaccine induced antibodies, Trastuzumab (10 μg/ml), or saline and an MTT assay was used to determine proliferation. Statistical analysis comparing samples to the vehicle alone treatment: * trastuzumab p= 0.010; ** HER2-VIA p> 0.0001.

(a) HER2-specific T cell responses in vaccinated mice. Pooled splenocytes obtained from C57BL/6 mice (n=8 per treatment group) 14 days after injection of Ad-HER2-ki, Ad-LacZ, or saline, were tested in an interferon-gamma ELISPOT and the number of responding T cells (+/− std deviation) was determined. The number of responding cells per 500,000 splenocytes in response to the antigen stimulus (x-axis) is shown. (b) HER2-specific T cell responses in vaccinated mice. Serum obtained from the same mice as in A at 14 days post injection of Ad-HER2-ki or Ad-LacZ was mixed with the listed cell lines shown on the x-axis. Binding of anti-HER2 antibodies was detected by ELISA. Absorbance was measured at 660 nm. (c) Polyclonal anti-HER2 Vaccine Induced Antibodies (Ad-HER2-VIA) mediates CDC in vitro. Pooled serum (8 mice per group) from C57BL/6 mice immunized with Ad-HER2-ki or Ad-LacZ, or, trastuzumab (10 μg/ml), was mixed with HER2 overexpressing cell lines (BT474 and SKBR3) or a HER2 negative cell line (MDA-231) and then rabbit complement was added. The percentage lysis of the cells (+/− std dev) was determined by Chromium release assay. (d) Polyclonal anti-HER2 Vaccine Induced Antibodies (Ad-HER2-VIA) mediates ADCC in vitro. NK cells derived from C57BL/6 mice were cultured with the anti-HER2 vaccine induced antibodies, anti-LacZ vaccine induced antibodies, or trastuzumab, and the cell line 4TI-HER2. The percentage lysis of the cells (+/− std dev) was determined by Chromium release assay. (e) Anti-proliferative effect of HER2-vaccine induced antibodies on HER2+ SKBR3 proliferation. HER2+ SKBR3 cells were cultured with HER2-vaccine induced antibodies, LacZ-vaccine induced antibodies, Trastuzumab (10 μg/ml), or saline and an MTT assay was used to determine proliferation. Statistical analysis comparing samples to the vehicle alone treatment: * trastuzumab p= 0.010; ** HER2-VIA p> 0.0001.