[Activating effects of protein transduction domain mediated BCR/ABL protein on CML T cells].
OBJECTIVE
To study the activating effect of protein transduction domain (PTD) mediated BCR/ABL protein on T cells from CML patients.


METHODS
The 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).


RESULTS
The 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).


CONCLUSIONS
By 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.
Liu
Qiang
Q
Department of Hematology of Tangdu Hospital, Fourth Military Medical University, Xi-an 710038, China.
Liang
Ying-min
YM
Zheng
Jin
J
Li
Xue-yi
XY
Hao
Miao-wang
MW
Liu
Li
L
Chen
Ren-an
RA
D000293:Adolescent; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D005260:Female; D005434:Flow Cytometry; D016044:Fusion Proteins, bcr-abl / Q000235:genetics* / Q000378:metabolism; D015696:Gene Products, tat / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood; D008297:Male; D008875:Middle Aged; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D015704:CD4 Antigens; D016827:CD8 Antigens; D016044:Fusion Proteins, bcr-abl; D015696:Gene Products, tat; D037181:Lectins, C-Type; D011993:Recombinant Fusion Proteins; D000099192:CD69 Antigens