Activating effects of protein transduction domain mediated BCR/ABL protein on CML T cells.
- Author:
Qiang LIU
1
;
Ying-min LIANG
;
Jin ZHENG
;
Xue-yi LI
;
Miao-wang HAO
;
Li LIU
;
Ren-an CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Adolescent; Adult; Aged; Amino Acid Sequence; Antigens, CD; analysis; Antigens, Differentiation, T-Lymphocyte; analysis; CD4 Antigens; analysis; CD8 Antigens; analysis; Dose-Response Relationship, Drug; Female; Flow Cytometry; Fusion Proteins, bcr-abl; genetics; metabolism; Gene Products, tat; genetics; metabolism; Humans; Lectins, C-Type; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; blood; Male; Middle Aged; Recombinant Fusion Proteins; immunology; metabolism; pharmacology; T-Lymphocytes; drug effects; immunology
- From: Chinese Journal of Hematology 2003;24(12):644-647
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the activating effect of protein transduction domain (PTD) mediated BCR/ABL protein on T cells from CML patients.
METHODSThe plasmid containing PTD and b3a2 bcr/abl of CML was constructed by genetic engineering and expressed in E. coli. The peripheral blood mononuclear cells from CML patients were stimulated in vitro with purified PTD-BCR/ABL protein and the expression of the early activation antigen CD(69) on CD(8)(+) and CD(4)(+) T cells was detected by flow cytometry (FCM).
RESULTSThe optimal concentration of PTD-BCR/ABL protein for activating CD(8)(+) T cells in vitro was 100 micro g/ml, CD(69) expression peaked in three days stimulation. CD(8)(+) T cells were activated in 10 of 15 CML patients, the expression rate of CD(69) was (15.01 +/- 3.75)%. CD(4)(+) T cells were activated in 4 of 15 patients, the expression rate of CD(69) was (10.32 +/- 3.08)%. Both CD(8)(+) and CD(4)(+) T cells were activated simultaneously in 3 of them. However, neither CD(4)(+) nor CD(8)(+) T cells was activated by stimulation with BCR/ABL protein in all 15 specimens, the expression rate of CD(69) on CD(8)(+) and CD(4)(+) T cells was (1.36 +/- 0.31)% and (1.41 +/- 0.43)%, respectively. There was no difference compared with that of PBS control group (P > 0.05).
CONCLUSIONBy using a PTD-mediated antigen delivering system, exogenous BCR/ABL protein can be delivered into APC, processed and presented onto surface of APC to activate Ag-specific CD(8)(+) and CD(4)(+) T cells in vitro.
