Research Of The Effect Of Tuberculin Pure Protein Derivative On ΓΔT Cell

Objectives:To investigate the effect of tuberculin pure protein derivative(PPD) onγδT cell.Methods:1. Mononuclear cells were centrifugalized by Ficoll density gradient centrifugation method from human peripheral blood, and T cells were obtained with nylon wool column method.2.The T cells were treated and stimulated by BCG and PPD to increase the number ofγδT cells, and theγδT cells were separated and purified via Flow Cytometry.3. The phenotype ofγδT cells as antigen-presenting cells was measured by Flow Cytometry. After separated from PBMC with adherent method, the mononuclear cells were induced to differentiate to mature dendritic cells (DC) which were detected 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 PPD and been incubated together with Mtb-Sag for 2 hours;④the mature dendritic cells incubated 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 PPD had increased, the levels of theγδT cells activated by PPD were similar to those ofγδT cells activated by BCG.2.In both of the two groups, the total number of T cells and CD4~+ T cells had increased, the number of PPD group was similar to BCG group,and lower than DC group.Conclusions:PPD could stimulate the expression of CD80,CD86 and HLA-DR of human peripheral bloodγδT cells, which indicated that PPD involved in the activation ofγδT cell.