title: [Role of cytokines and gene expression characteristics in cultured lymphocytes ex vivo for adoptive immunotherapy].
abstract: Different cytokines are needed in the course of culturing cells to do adoptive immunotherapy. This study was aimed to investigate the differentiation directions of lymphocytes and related gene expression characteristics after combined stimulation of lymphocytes by different cytokines or EBV antigen peptide combined with cytokines. The experiment was divided into 4 groups. The levels of total T lymphocytes (CD3(+)), T helper lymphocytes (CD3(+)CD4(+)), cytotoxic T-lymphocyte (CD3(+)CD8(+)), memory T cells (CD3(+)CD8(+)CD45RO(+)), naive T cells (CD3(+)CD8(+)CD45RA(+)), Th2 cells (CD3(+)CD30(+)), B cells (CD19(+)), NK cells (CD56(+)), naive T regulatory cells (CD4(+)CD25(+)), precise T regulatory cells (CD4(+)CD25(+)FOXP3(+)) were detected by flow cytometry. The expression levels of house-keeping gene (mad1, pten), T helper cells transcriptional regulatory gene t-bet (Th1), gata3 (Th2), cytokine IFN-γ(Th1), IL-4(Th2) were detected by using RT-PCR. The results showed that CTL in EBV polypeptide group were dominant cells with certain clinical effects. Comparison of result of EBV polypeptide group with other 3 different cytokine stimulating groups demonstrated that EBV antigen peptide had much more effects on stimulating CTL generation. The expression of IFN-γ gene was significantly increased; the T helper differentiation-related gene t-bet, gata3 also increased evidently, while expression change of house-keeping gene mad1 and pten were not evident. Addition of different cytokines and antigen peptides in culture may be much more effective on stimulating CTL generation. It is concluded that specific CTL can be obtained by using the lymphocytes co-cultured with EBV and cytokines, and the different cytokines play different roles in cell differentiation.
authors.affiliation: Department of Hematology, Peking University First Hospital, Beijing 100034, China.
authors.forename: Gai-Ling
authors.initials: GL
authors.lastname: Zhang
authors.forename: Xiu-Yan
authors.initials: XY
authors.lastname: Tao
authors.forename: Zhao-Li
authors.initials: ZL
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Hong-Xing
authors.initials: HX
authors.lastname: Liu
authors.forename: Peng
authors.initials: P
authors.lastname: Cai
authors.forename: Ding-Fang
authors.initials: DF
authors.lastname: Bu
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
mesh_terms: D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive; D018655:Lymphocyte Count; D054391:Lymphoma, Extranodal NK-T-Cell / Q000235:genetics* / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D016207:Cytokines; D019309:Epstein-Barr Virus Nuclear Antigens