Aim: To study the activation effect of BCR/ABL antigen on T cells from CML patients mediated by protein transduction domain (PTD).
Methods: The fused plasmid containing PTD gene and b3a2 bcr/abl gene of CML was constructed by genetic engineering technique and was expressed in E.coli. The PBMCs from CML patients were stimulated in-vitro with purified PTD-BCR/ABL antigen and then expression of the activation antigen CD25 on CD8(+) and CD4(+) T cells after stimulation was detected by flow cytometry (FCM).
Results: After stimulation with 100 mg/L of PTD-BCR/ABL antigen (final concentration) for 4 days in-vitro, CD8(+) T cells were activated in 5 of 10 CML patients and CD4(+) T cells were activated in 2 of 10 patients. Both CD8(+) and CD4(+) T cells were activated simultaneously in one of them. However, neither CD4(+) nor CD8(+) T cells was activated in BCR/ABL antigen stimulation group as control.
Conclusion: Using a PTD-mediated antigen transduction system, exogenous BCR/ABL antigen can be transferred into APCs and be processed and presented onto surface of APCs to activate Ag-specific CD8(+) and CD4(+) T cells in-vitro. The strategy outlined in this paper may provide a new approach for priming Ag-specific CD8(+) and CD4(+) T cells in-vitro and immunotherapy of CML.