Abstract
A direct positive panning technique was developed in order to achieve highly purified CD4+ and CD8+ cells. Fresh peripheral blood mononuclear cells or tumor-infiltrating lymphocytes derived from bulk cultures were applied to modified polystyrene surfaces to which anti-CD4 or anti-CD8 monoclonal antibodies were covalently attached. Adherent cells were allowed to grow in the original flask and were then harvested and cultured in IL-2-containing medium. This positive immunoselection technique resulted in CD4+ and CD8+ cell subsets with high cell viability and a high degree of purity. In several samples, the isolated cell subsets were subsequently subjected to a negative immunomagnetic bead selection in order to remove reciprocal cell subset contamination or double-positive CD4+/CD8+ cells. The isolated cells maintained their ability to proliferate, kept their phenotypic profiles, and remained functionally intact after long-term growth in culture without the further addition of mitogenic or allogeneic cell stimulation. This approach is a simple, rapid, and reproducible technique that might be useful on a large scale to isolate and to grow T-cell subsets for research and for clinical use.
Original language | English (US) |
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Pages (from-to) | 463-474 |
Number of pages | 12 |
Journal | Journal of Biological Response Modifiers |
Volume | 9 |
Issue number | 5 |
State | Published - Oct 1990 |
Keywords
- CD4 and CD8 cells
- Negative immunomagnetic selection
- Positive panning
- T-cell subsets
ASJC Scopus subject areas
- Immunology
- Pharmacology
- Cancer Research