| 摘要 |
<p>A pharmaceutical preparation, comprising a cell-cycle regulatory protein, and/or a nucleic acid encoding that protein, for immunization against carcinoma and its preliminary stages, is new. - ACTIVITY : Cytostatic. Peripheral blood lymphocytes were isolated from an HLA-A0201 positive sample by density centrifugation and T lymphocytes were separated from monocytes and B lymphocytes using antibodies coupled to magnetic beads (CD11, CD16, CD19, CD36 and CD56) (T cell isolation kit, Milteny, Germany). Peptides of p16 were identified using a software system (NIH bioinformation service) including the following peptides ValMetMetMetGlySerAlaArgVal, ValLeuHisArgAlaGlyAlaArgLeu, TheLeuThrArgProValHisAspAla, LeuLeuHisGlyAlaGluProAsnCys, SerMetGluProSerAlaAspMetLeu, MetMetGlySerAlaArgValAlaGluLeu, LeuLeuLeuHisGlyAlaGluProAsnCys, GlyValMetMetMetGlySerAlaArgVal. The isolated T cells were incubated with T2 cells and a mixture of the 9mer (a) or 10mer (b) peptides. The T cells were re-stimulated weekly over a six week period. In each case 10 7> T-cells were co-cultivated with 2x10 6> peptide-loaded T2 cells in 24 well plates. Reaction against the peptide-loaded T2 cells was determined weekly using an interferon (IFN)-lambda enzyme linked immunosorbent assay (ELISA). On day 28 reaction against mixture (a) was observed with the main reaction being against peptide ValMetMetMetGlySerAlaArgVal (1000 specific cells per million cells), and a weaker reaction against mixture (b) with peptide MetMetGlySerAlaArgValAlaGluLeu being the highest (600 specific cells per million cells). Thus it is clear that CD8 +> T-cells can be stimulated against p16. These activated T-cells were incubated with human leukocyte antigen (HLA) A0201+ Caski cervix carcinoma cells which over-express p16, and with colon carcinoma line SW480 which do not over-express p16. The Caski cells were lysed, whilst the controls were not. - MECHANISM OF ACTION : Vaccine; gene therapy.</p> |
