Dear experts, This is not exactly a flow question but maybe someone can help me. I am trying to generate the CD8 cell lines specific for an antigenic peptide. I have put into culture fresh CD8+ cells from a healthy donor that I stimulate weekly with homologous PBMC (pulsed with a peptide (10ug) for 3 hours and irradiated) along with Il-2 and Il-7. I haven't seen the peptide-specific cytotoxic lymphocyte (checked with HLA tetramers) till the 6th week of culture. I would be grateful for any suggestion what I can change or any reliable protocol. Thanks. Michal -- Michal Abel, MD, PhD Laboratory of Clinical Immunology INSERM U25 Necker Hospital, 161 Rue de Sèvres. Paris tel: 33 1 42 19 28 87 fax: 33 1 44 49 53 74
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