Quantitation, Phenotypic And Functional Analyses Of HCMV Specific Cytotoxic T Lymphocytes In Peripheral Blood Monocyte Lymphocytes From Healthy Chinese HLA-A2～+ Donors

AIM: To optimize tetramer staining condition using HLA-A*0201 tetramer(A2-NLV tetramer) loaded with NLV peptide (pp65_495-503 derived from structural protein pp65 of human cytomegalovirus,and to investigate the phenotypic and functional characteristics and TCR Vβ usage of HCMV-specific CTL cells in healthy young and elderly Chinese donors.METHODS:1. Peripheral blood from HLA-A2⁺ donors was first stained with A2-NLV tetramer/PE under different conditions and then labeled with anti-CD3-FITC and anti-CD8-APC. The stained samples were analyzed with flow cytometry to find out the optimized staining condition.2. Whole blood from healthy HLA-A2⁺ young and elderly Chinese donors were stained with a combination of the tetramers and antibodies for CD28 and CD57 and were analyzed by three-color flow cytometry.3. Perforin expression was analyzed with flow cytometry by using A2-NLV tetramer staining together with intracellular staining without stimulation, while γ-interferon...