The Influence of three different activators on CD69 expression of CD3+ lymphocytes

Objectives To study the influence of three different activators on CD69 expression of CD3+ lymphocytes. Methods Peripheral blood mononuclear cells(PBMCs) were separated from the blood of healthy contributors and stimulated with CD3 monoclonal antibodies (CD3mAb, group A), phorbol 12-myristate 13-acetate plus ionomycin (PMA+ IM, group B), or low molecular peptide antigen of mycobacterium tuberculosis (Mtb-Ag, group C). The dynamic expression of CD69 molecule on CD3+ T cells and γδΤ cells was detected by flow cytometric analysis at 0, 6, 12, 24, 48, and 72 hours after stimulation. Results The level of CD69 expression on CD3+ T cells in group A reaches the summit of 56.2% at 24 h. It reaches the summit of 99.5% at 6 h in group B and 16% at 24 h in group C respectively (F=322.528, P<0.001, pairwise comparison: P<0.001). The level of CD69 expression on γδΤ cells in group A reaches the summit of 55.1% at 24 h. It reaches the summit of 99.9% at 6 h in group B and 75.2% at 24h in group C separately(F=21.170, P<0.001, pairwise comparison: P<0.001 when A was compared with B and B compared with C, P=0.646 when C was compared with A). Conclusions Among the three activators, PMA+ IM is most efficient in activating CD3+ T cells and γδΤ cells. It can induce the highest level of CD69 expression in the shortest time period. Key words: CD3mAb; PMA; IM; Mtb-Ag; CD3+ T cell; CD69 molecule