| Objectives:To research the antigen-presenting cells'phenotype and functions expressed by human peripheral bloodγδT cells stimulated by Bacille Calmette-Guerin (BCG) and early secreted antigenic target-6 (ESAT-6).Methods:1.Centrifugalized mononuclear cells from human peripheral blood with the Ficoll density gradient centrifugation method, and the T cells were gained by the nylon wool column method.2.The T cells were treated and stimulated by BCG and ESAT-6 to increase the number ofγδT cells, and theγδT cells were separated and purified via Flow Cytometry.3.The antigen-presenting cells'phenotype ofγδT cells were detected by Flow Cytometry. The mononuclear cells had been separated from PBMC by adherent methods, and induced to differentiate into mature dendritic cells (DC), then the maturity of DCs were estimated by Flow Cytometry. 4.The T cells marked with CFSE were cultured in four groups:①The secreted antigen of Mycobacterium tuberculosis (Mtb-Sag);②theγδT cells activated by BCG and been incubated together with Mtb-Sag for 2 hours;③theγδT cells activated by Esat-6 and been incubated together with Mtb-Sag for 2 hours;④the mature dendritic cells which had been incubated together with Mtb-Sag for 2 hours.At last detected the proliferation of the T cells and the CD4 + T cells.Results:1.The CD80, CD86 and HLA-DR expression levels of theγδT cells activated by BCG and ESAT-6 had increased, but the levels of theγδT activated by Esat-6 were significantly higher than those ofγδT activated by BCG.2.In both of the two groups, the total number of T cells and CD4 + T cells had increased, however the number of Esat-6 group was much higher, as has been shown when the T cells were cultured with mature dendritic cells.Conclusions:The human peripheral bloodγδT cells activated by BCG and ESAT-6 have the antigen-presenting cells'phenotype and antigen presentation functions, and the antigen presentation functions are much stronger in the Esat-6 group. |
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