Killing effect of T-lymphocytes after activated by dendritic cells loaded with MGC-803 cells with overexpressed cell cycle related transcription factor 1

Abstract

Objective To study the methods of induction of dendritic cells (DC) which derived from the peripheral blood of health adults and further to study the anti-tumor of kidney effect activated by DC.Methods With 30 μg/L recombinant human interleukin-4 (rhlL-4),100 pg/L recombined human GM-CSF (rhGM-CSF),50 μg/L recombinant human tumor necrosis factor-α (rhTNF-α) andl00 μg/L lipopolysaccharide (LPS) make nuclear cell become the DC,Through the morphological and flow cytometric identification of DC,cell cycle related transcription factor 1 (E2F-1) MGC-803 made antigen sensitization stable strains after DC with cell counting kit-8 (CCK-8) method to detect T lymphocyte proliferation in DC:T (1:5 ; 1:10 ; 1:20 ; 1:40).The killing effect of cytotoxic T lymphocyte (CTL) by DC loaded detect the Absorbance of different group by methyl thiazol tetrazolium (MTT) assay and calculate T lymphocytes stimulating index in T cell:MGC-803 (10:1,20:1,50:1).Results Morphology and Flow cytometric show the induction of DC cells mature features obvious and overexpressed E2F-1 antigen sensitization DC can promote T lymphocyte proliferation and express E2F-1 group T lymphocytes SI value (4.42 ± 0.23)was significantly higher than the two other groups [empty carrier group (2.95 ± 0.53) and blank control group (3.01 ±0.68),P ＜0.05].A E2F-1 antigen expression sensitization of DC induction to cancer of the stomach CTL MGC-803 cell damage rate (65.00 ± 8.24) ％ was significantly higher than other groups,[DC-CTL (45.25 ±7.8) ％,MGC-803 group (41.83 ±4.79)％,empty carrier group (31.16 ±11.96) ％,blank control group (33.25 ± 8.38) ％,P ＜ 0.05],express E2F-1 antigen group T lymphocytes secretion interferon (IFN)-γ amount (571.03 ± 5.96) ng/L is much higher than in other groups [(382.85 ±4.942) ng/L,(390.13 ±7.06) ng/L,(356.03 ±5.36) ng/L,P＜0.05].Conclusion Over expression E2F-1 MGC-803 antigen sensitization to DC in vitro can increase T lymphocytes added value and strengthen the T lymphocytes to kill MGC-803 cell. Key words: Dendritic cells; Transcription factor; T-lyphocytes; Immune killer; Interferon-γ