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Effect of Immune Response Induced by APL Cells and PML-RARα Vaccine in Vitro and in Vivo

Author: ZuoDongZhi
Tutor: LiYangQiu
School: Jinan University
Course: Hematological Diseases
Keywords: Acute promyelocytic leukemia PML-RARα T cell receptor (TCR) Gene vaccine
CLC: R733.71
Type: Master's thesis
Year: 2008
Downloads: 32
Quote: 0
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Abstract


Objective: To analyze the acute promyelocytic leukemia (APL)-associated antigen-specific T cells induced immune response; understanding of the PML-RARα fusion gene vaccine in vivo immune effects, provided the experimental basis for conducting the DNA vaccine in the treatment of the APL. Methods: (1) the use of RT-PCR amplification 9 APL patients 29 TCR Vα subfamily genes complementarity determining region 3 (CDR3), to understand the distribution of TCR Vα subfamily T cells; positive products were further Genescan analysis CDR3 length clones. Peripheral blood of 10 normal subjects as controls. (2) using the mixed lymphocyte tumor cell culture (MLTC) method, the use of APL cells in vitro human umbilical cord blood T-cell proliferation, proliferation characteristics analysis of umbilical cord blood TCR Vα and Vβ subfamily application LDH method analysis of induced proliferation of T cells APL cell specific cytotoxicity. (3) successfully constructed recombinant plasmid (pIRES-PML-RARα-hIL-2) in mice immunized with separate injection site muscle, using RT-PCR and immunohistochemical staining for detection of target gene transcription in mouse skeletal muscle ELISA assay serum INF-γ production and expression;; LDH release assay detection of mouse spleen cells specific CTL killing rate for APL cell line NB4; indirect immunofluorescence assay serum anti-PML-RARα antibody ; probe method is real-time quantitative PCR detection of T-cell receptor excision DNA circles (TRECs) to understand the level of mice the thymus naiveT cells. Results: (1) most APL blood T cells can be detected to seven the TCR Vα Asian family, a higher frequency of occurrence to Vα3 and Vα19; clonal proliferation of T cells can be detected in 8 patients, to Vα3, Vα26 and Vα27 clonal T-cell frequency (3/8). (2) after the APL cells stimulated proliferation of cord blood T cells to express the the part Vα and Vβ the pedigree gene, and appear in of Vα10, Vβ5 and Vβ15 clonal proliferation trend after the induction of T cell destruction of APL cells are higher than induced group. (3) the mouse injection site muscle detected PML-RARα and hIL-2 gene transcription can be detected hIL-2 protein expression. Seven weeks after immunization, the serum content of INF-γ, anti-PML-RARα antibody of TRECs and spleen CTL cells in for a NB4 killing rate higher than that of the control group. Conclusion: APL cell associated antigens can induce T cell-specific immune response, performance utilization and clonal proliferation of TCR Vα and Vβ subfamily advantage, and APL cell specific cytotoxic. PML-RARα series constructed recombinant plasmid and induced humoral and cellular immune responses, with the basic characteristics of the DNA vaccine.

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia > Acute leukemia
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