Aim: To explore the method of selective culture of gamma delta T cells in peripheral blood and bronchoalveolar lavage fluid(BALF) for obtaining pure gamma delta T cell subset.
Methods: A novel attack-panning method for selectively culturing gamma delta T cells was set up. The peripheral blood mononuclear cells (PBMCs) and BALF were isolated by Ficoll-hypaque density gradient centrifugation (n=10). alpha beta T cells in PBMCs were depleted by complement-dependent-cytotoxicity(CDC) after the monocytes/macrophages were removed by adherence. The gamma delta T cells in PBMCs were cultivated selectively using anti-TCR gamma delta monoclonal antibody and IL-2. The proliferation of gamma delta T cells were observed by plotting growth curve. The purity of gamma delta T cells were detected by flow cytometry and immunohistochemistry.
Results: Stimulated by anti-TCR gamma delta mAb and IL-2, gamma delta T cells of peripheral blood and BALF could proliferate for a few days. The purity of gamma delta T cells obtained by Attack-panning method was 81%-99%.
Conclusion: The attack-panning method can get pure gamma delta T cells from peripheral blood and BALF.