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Updated: Aug 17, 2026
Preparation of CD4+ T Cells for Analysis of GD3 and GD2 Ganglioside Membrane Expression by Microscopy
Published on: November 8, 2016
[Exploration of ganglioside GM1 as an activation marker on T cells]
1Department of Immunology, Bengbu Medical College, Bengbu 233003, China.
Aim:
To compare and analyze the patterns of ganglioside GM1 and CD69 expressions on activated gammadeltaT cells and CD3(+) T cells from human peripheral blood.
Methods:
PBMCs were stimulated in vitro with anti-CD3 mAb or Mycobacterium tuberculosis antigen (Mtb-Ag). In some experimental groups, PBMCs were pretreated with different inhibitors of signal transduction pathway before stimulation. The expressions of GM1 or CD69 on gammadeltaT and CD3(+) T cells at different time points after stimulation were measured by flow cytometry (FCM).
Results:
GM1 appeared on gammadeltaT cells at 30 min after stimulation with Mtb-Ag and retained a higher level with the lapse of time; whereas CD69 expression appeared at 3 h after stimulation and reached peak at 24 h, and then decreased gradually to the resting level on day 6 after stimulation. The expressions of CD69 and GM1 on CD3(+) T cells stimulated with anti-CD3 mAb were similar to those on gammadeltaT cells. The expression of GM1 on gammadeltaT cells was inhibited by pretreatment with PD98059 or LY294002. PMA increased the expression of GM1.
Conclusion:
GM1 can serve as a novel marker of activated T cells. During the activation of T cells, the dynamic expression of GM1 was different from that of CD69, a very early activation antigen. The Ras-Erk signal transduction pathway and PI3K and PKC might be involved in the expression of GM1.
