Zhang Xueyan, Zhang Yanwei, Xu Jianlin, Wang Huimin, Zheng Xiaoxuan, Lou Yuqing, Han Baohui
Department of Pulmonary, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai 200030, China.
J Cancer. 2018 Jan 1;9(2):367-374. doi: 10.7150/jca.20821. eCollection 2018.
The present study investigated whether antigen presentation of the Oct4 and Sox2 peptides by CD154-activated B lymphocytes can enhance the killing effect of CD8 cytotoxic T lymphocytes (CTLs) on lung stem-like cancer cells (SLCCs). The CTLs were generated using an accelerated co-cultured dendritic cells (DC) (acDC) assay by incubating human peripheral blood mononuclear cells (PBMCs) from non-small-cell lung cancer patients with antigen peptides of Oct4 and Sox2 in the presence of several DC-activating agents. CD154 NIH3T3 cells prepared by CD154 lentiviral transfection were used as feeder layer to activate primary B cells (CD19) obtained from PBMCs. Activated B cells were co-cultured with CTLs to present antigen peptides of Oct4 and Sox2. CTLs co-cultured with activated B cells were evaluated for the levels of secreted inflammatory cytokines using ELISA. In addition, the killing effect of the CTLs on SLCCs derived from cisplatin-resistant strain of human lung cancer cell line PC9 was evaluated by flow cytometry using CFSE labeling of the target cells. After the acDC assay, the PBMCs exhibited a significant (p<0.01) increase in the population of CD8/CD3 cells, indicating successful preparation of CTLs. The primary B cells cultured on the CD154 NIH3T3 feeder layer resulted in significant (p<0.01) increase in the proportions of population expressing CD80, CD86, or HLA-A, indicating successful activation of the B cells. The co-culture of CTLs with CD154-activated B cells presenting the Oct4 and Sox2 peptides caused significant increase in the levels of secretory inflammatory cytokines and exhibited enhanced killing of the SLCCs derived from cisplatin-resistant PC9 cells. Antigen presentation of the Oct4 and Sox2 peptides by CD154-activated B cells can enhance the killing effect of CTLs towards lung SLCCs.
本研究调查了CD154激活的B淋巴细胞呈递Oct4和Sox2肽的抗原是否能增强CD8细胞毒性T淋巴细胞（CTL）对肺干细胞样癌细胞（SLCC）的杀伤作用。通过将非小细胞肺癌患者的人外周血单个核细胞（PBMC）与Oct4和Sox2的抗原肽在几种DC激活剂存在下孵育，使用加速共培养树突状细胞（DC）（acDC）试验生成CTL。通过CD154慢病毒转染制备的CD154 NIH3T3细胞用作饲养层，以激活从PBMC获得的原代B细胞（CD19）。将激活的B细胞与CTL共培养以呈递Oct4和Sox2的抗原肽。使用ELISA评估与激活的B细胞共培养的CTL分泌的炎性细胞因子水平。此外，通过使用CFSE标记靶细胞的流式细胞术评估CTL对源自人肺癌细胞系PC9顺铂耐药株的SLCC的杀伤作用。在acDC试验后，PBMC中CD8/CD3细胞群体显著（p<0.01）增加，表明成功制备了CTL。在CD154 NIH3T3饲养层上培养的原代B细胞导致表达CD80、CD86或HLA-A的群体比例显著（p<0.01）增加，表明B细胞成功激活。CTL与呈递Oct4和Sox2肽的CD154激活的B细胞共培养导致分泌性炎性细胞因子水平显著增加，并表现出对源自顺铂耐药PC9细胞的SLCC的杀伤增强。CD154激活的B细胞呈递Oct4和Sox2肽的抗原可增强CTL对肺SLCC的杀伤作用。
