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. 2005 Apr 5;92(8):1421–1429. doi: 10.1038/sj.bjc.6602526

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Copyright © 2005 Cancer Research UK

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Evaluation of vaccine-induced immune responses. BALB/c mice were immunised in separate experiments by s.c. injection of Tc-ErbB2/Th-HA liposomes (white bars), Tc-ErbB2 liposomes (black bars), or peptide-free liposome carrier (A) or PBS (B) (hatched bars) on days 0 and 14. (A) To investigate induction of ErbB2-specific CTLs, 5 days after vaccination splenocytes were isolated and stimulated with synthetic ErbB2 peptide before flow cytometric analysis with anti-CD8α and anti-IFN-γ antibodies. Absolute numbers of CD8⁺ IFN-γ⁺ splenocytes are indicated (mean values from five animals per group). (B) Sera taken at day 21 after the first vaccination were tested for the presence of peptide epitope-specific antibodies by ELISA. ErbB2-derived Tc peptide was coated on ELISA plates, before sera were added. Peptide-specific antibodies were detected by HRP-conjugated secondary anti-mouse IgG, followed by addition of HRP-substrate (ABTS) and measuring of the absorbance at 405 nm in an ELISA reader.