Generation of populations of antigen-specific cytotoxic T cells using DCs transfected with DNA construct encoding HER2/neu tumor antigen epitopes

BMC Immunology

Methodology Article

Julia Lopatnikova¹  Maria Kuznetsova¹  Julia Khantakova¹  Sergey Sennikov¹  Rinat Maksyutov²  Amir Maksyutov³ 

[1] Federal State Budgetary Scientific Institution “Research Institute of Fundamental and Clinical Immunology”, Yadrintsevskaya str., 14, 630099, Novosibirsk, Russia;State Research Center of Virology and Biotechnology “VECTOR”, 630559, Koltsovo, Novosibirsk Region, Russia;State Research Center of Virology and Biotechnology “VECTOR”, 630559, Koltsovo, Novosibirsk Region, Russia;Federal State Budgetary Scientific Institution “Research Institute of Fundamental and Clinical Immunology”, Yadrintsevskaya str., 14, 630099, Novosibirsk, Russia;

关键词: Cytotoxic T cells;    CTLs;    Antigen-specific cells;    Dendritic cells;    HER2/neu;    Tumor-associated antigen;    Antitumor immune response;   

DOI  :  10.1186/s12865-017-0219-7

 received in 2017-04-04, accepted in 2017-06-16,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundRecent fundamental and clinical studies have confirmed the effectiveness of utilizing the potential of the immune system to remove tumor cells disseminated in a patient’s body. Cytotoxic T lymphocytes (CTLs) are considered the main effectors in cell-mediated antitumor immunity. Approaches based on antigen presentation to CTLs by dendritic cells (DCs) are currently being intensively studied, because DCs are more efficient in tumor antigen presentation to T cells through their initiation of strong specific antitumor immune responses than other types of antigen-presenting cells. Today, it has become possible to isolate CTLs specific for certain antigenic determinants from heterogeneous populations of mononuclear cells. This enables direct and specific cell-mediated immune responses against cells carrying certain antigens. The aim of the present study was to develop an optimized protocol for generating CTL populations specific for epitopes of tumor-associated antigen HER2/neu, and to assess their cytotoxic effects against the HER2/neu-expressing MCF-7 tumor cell line.MethodsThe developed protocol included sequential stages of obtaining mature DCs from PBMCs from HLA A*02-positive healthy donors, magnet-assisted transfection of mature DCs with the pMax plasmid encoding immunogenic peptides HER2 p369–377 (E75 peptide) and HER2 p689–697 (E88 peptide), coculture of antigen-activated DCs with autologous lymphocytes, magnetic-activated sorting of CTLs specific to HER2 epitopes, and stimulation of isolated CTLs with cytokines (IL-2, IL-7, and IL-15).ResultsThe resulting CTL populations were characterized by high contents of CD8+ cells (71.5% in cultures of E88-specific T cells and 90.2% in cultures of E75-specific T cells) and displayed strong cytotoxic effects against the MCF-7 cell line (percentages of damaged tumor cells in samples under investigation were 60.2 and 65.7% for E88- and E75-specific T cells, respectively; level of spontaneous death of target cells was 17.9%).ConclusionsThe developed protocol improves the efficiency of obtaining HER2/neu-specific CTLs and can be further used to obtain cell-based vaccines for eradicating targeted tumor cells to prevent tumor recurrence after the major tumor burden has been eliminated and preventing metastasis in patients with HER2-overexpressing tumors.

【 授权许可】

CC BY   
© The Author(s). 2017