Development of a lung cancer vaccine by transfecting dendritic cells with rAAV/CEA
10.3969/j.issn.1673-4254.2014.04.10
- VernacularTitle:rAAV/CEA转染树突状细胞制备肺癌疫苗
- Author:
Changxuan YOU
1
;
Xiaotao QIAN
;
Yuan HE
;
Liu YONG
;
Hermonat L PAUL
Author Information
1. 南方医科大学南方医院肿瘤科
- Keywords:
carcino-embryonic antigen;
adeno-associated virus;
lung cancer;
vaccine
- From:
Journal of Southern Medical University
2014;(4):487-491
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells (DCs) with adeno-associated virus vector carrying carcino-embryonic antigen gene (rAAV/CEA). Methods Adherent cells (monocytes) isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide (control). The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with 3H-thymidine uptake, and the expression of IL-4, IFN-γ, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes (CTL) was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by 51Cr releasing assay. Results The DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro. Conclusion It is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/CEA.
