Functional molecular expression and frequency of CD8+ T lymphocytes

The frequency of CD8+ T lymphocytes of all patients in leukomonocytes, CD69+CD8+ T lymphocytes, CD178+CD8+ T lymphocytes, and the counts of CD69 and CD178 expressed on CD8+ T lymphocytes were detected by flow cytometry (FACS Caliburflow Cytometer, USA). It was performed as follow steps: 1) using monoclonal Antibodies (mAbs) of CD3-peridinin chlorophyll protein (PerCP), CD8-human antigen presenting cells (APC), CD69-phycoerythrin (PE), and CD178-PE (BD Biosciences, Cowley, UK) incubated 100μl of whole peripheral blood samples in the dark for 20 minutes; 2) incubated with 2ml fluorescently activated cell classifier (FACS) lysate for 5 mins. 3) after centrifuged at 300×g 5 minutes and aspirated the supernatant, 2ml phosphate buffered saline (PBS) was added and vortex lightly; 4) after centrifuged at 300×g 5 minutes and aspirated the supernatant, the sample was re-suspended with 200μl of PBS and finally analyzed by FACS flow cytometer. CD8+T lymphocytes were identified as monocytes which were in the subsequent showing CD3+/CD8+. CD3+CD8+CD69+ was considered to be the activated CD8+T lymphocytes, CD3+CD8+CD178+ was considered to be the apoptosis-inducing CD8+T lymphocytes.