Figure 7
Enumeration and reactivity detection of HBc18–27/HBs183–191-specific CD8+ T cells by AAPC-microplate. PBMCs from all subjects were detected by the AAPC-microplate method and traditional HLA-A2/HBc18–27/HBs183–191 dimers staining plus flow cytometry. (a) Magnetic separation of HBc18–27/HBs183–191-specific CD8+ T cells in the AAPC-microplate. Magnification is 400 × for each image. (b) The correlation coefficient between the AAPC-microplate method and flow cytometry as analyzed by two-tailed Pearson correlation. (c) The frequencies of HBc18–27/HBs183–191-specific CD8+ T cells in the PBMCs from all subjects as detected by the AAPC-microplate method and flow cytometry (FACS). (d) The reactivity of HBc18–27/HBs183–191-specific CD8+ T cells (AST) and whole T cells from the HLA-A2-positve patients with chronic Hepatitis B. HBc18–27/HBs183–191-specific CD8+ T cells retained in micro well after AAPC-bead sorting were further co-incubated with PHA for 24 hrs in the AAPC-microplate. Meanwhile, the PBMC samples from the patients were also co-incubated with PHA for 24 hrs without AAPC-bead sorting. IFN-γ local detection was performed by ELISPOT assay as described and the percentages of IFN-γ-secreting cells in the sorted AST cells and CD3+ T cell populations were calculated. Data are presented as mean ± SD.
