Journal: Journal of the National Cancer Institute

Article Title: Response of established human breast tumors to vaccination with mammaglobin-A cDNA.

doi: 10.1093/jnci/djh261

Figure Lengend Snippet: Fig. 1. Membrane stabilization of the HLA-A2 molecule by mammaglobin-A– derived candidate epitopes. Computer-assisted predicted HLA-A2–binding af- finities of the candidate epitopes were confirmed by membrane stabilization of the HLA-A2 molecule in T2 cells, a human T-B lymphoblastoid hybrid cell line deficient in transporter associated with antigen-processing proteins. T2 cells were incubated in the presence of each peptide in complete medium (see “Subjects and Methods”). After 24 hours, HLA-A2 expression was determined with flow cytometric analysis using the BB7.2 anti–HLA-A2 monoclonal anti- body (solid bars). T2 cells incubated in the presence of the GILGFVFTL influenza–derived epitope (Flu) were used as a positive control, and T2 cells incubated in the presence of HLA-A3–restricted PLLENVISK (Mam-A3.1) mammaglobin-A–derived epitope were used as negative control. Results are expressed as the mean fluorescence shift, which represents the difference be- tween the mean fluorescence obtained with T2 cells cultured in the presence of peptides and the mean fluorescence obtained with T2 cells cultured in the absence of peptides. Results are presented as the mean of four independent experiments performed in quadruplicate. Error bars represent upper 95% con- fidence intervals. Fig. 2. Epitope immunodominance of CD8 cytotoxic T lymphocytes (CTLs) from mammaglobin-A cDNA–vaccinated HLA-A2 /hCD8 mice. Five HLA- A2 /hCD8 mice were each vaccinated with either mammaglobin-A cDNA (solid bars) or empty vector (open bars). A) Immunodominance of the CD8

Article Snippet: The HLA-A2–binding ability of the peptides was confirmed by cell membrane stabilization of the HLA-A*0201 molecule in transporter associated with antigen processing (TAP)–deficient T2 cells, a human T-B lymphoblastoid hybrid cell line obtained from the American Type Culture Collection.

Techniques: Membrane, Derivative Assay, Binding Assay, Incubation, Expressing, Positive Control, Negative Control, Cell Culture, Plasmid Preparation