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In Vitro Immue Response of Cytotoxic T Lymphocyte Induced by Dendritic Cells Pulsed with Tumor Associated Antigen MUC4

Author: MengKai
Tutor: MiaoYi
School: Nanjing Medical University
Course: Department of General Surgery
Keywords: MUC4 Dendritic cells Cytotoxic T lymphocyte ELISPOT
CLC: R392
Type: Master's thesis
Year: 2008
Downloads: 42
Quote: 0
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Abstract


Objective: To investigate the tumor antigen of MUC4 HLA - A * 0201 - restricted CTL epitope peptide load dendritic cells induced by cytotoxic T lymphocytes (CTL) in vitro immune effector . Methods: induction of dendritic cells ( DC ) from peripheral blood mononuclear cells (PBMC) . Epitope prediction method prior to use to predict and synthesize five peptide molecules were pulsed mature DC, and to stimulate the HLA -A * 0201 to healthy human autologous CD8 ~~ T cells become CTLs . Week enzyme-linked immunosorbent spot assay (ELISPOT), to detect CTL IFN-γ secretion . Lactate dehydrogenase (LDH) release assay its killing effect on target cells . Results: The human PBMC in a variety of cytokines are induced to become DC . CTL induced by peptide P01204 different effector to target than the the specific killing P01204 pulse T2 cells ( 5:1 , 9.03 % ± 0.24 % , 19.62% ± 0.44 % in 10:1 , 20:1 , 27.93 % ± 2.22%) and MUC4 ~, HLA-A2 ~ HCT-116 cells ( 5:1 , 7.26% ± 0.18 % , 16.83 % ± 0.40 % in 10:1 , 20:1 and 25.23% ± 1.35 %), are higher than the negative control group ( P <0.05 ) , the number of cells able to produce IFN-γ (130.33 ± 6.58) was significantly higher than the negative control group ( P <0.05 ) . Conclusion: The tumor associated antigen MUC4 HLA - A * 0201 - restricted CTL epitope P01204 , could induce CTL response of the human peripheral blood mononuclear cells , the activation of CTL specific target cells induced apoptosis can secrete immunologically reactive substance .

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