?d T Cells Provide Protective Function in Highly Pathogenic Avian H5N1 Influenza A Virus Infection

Dong, Peng; Ju, Xiangwu; Yan, Yiwu; Zhang, Siya; Cai, Menghua; Wang, Huaishan; Chen, Hui; Hu, Yu; Cui, Lianxian; Zhang, Jianmin; He, Wei

Expansion and Isolation of γδ T Cells γδ T cells were expanded from human peripheral blood monocytes (PBMCs) as described previously (20) . Briefly, fresh PBMCs separated from the peripheral blood of healthy donors by Ficoll-Hypaque (Pharmacia, Sweden) density gradient centrifugation were grown in RPMI-1640 medium (Gibco BRL, USA) supplemented with 10% FCS and IL-2 (200 UI/ml) in 24well culture plates coated with an immobilized anti-pan-TCR γδ mAb (0.05 mg/ml, Immunotech, France). After 10 to 12 days of culture, the purity of the γδ T cells was more than 85%, as assessed by flow cytometric analysis. γδ T cells were further purified and concentrated to a purity of up to 98% using the TCRγ/δ+ T Cell Isolation Kit (Miltenyi Biotec, Germany) in accordance with the manufacturer's instructions.