干旱区生态环境知识资源中心(Arid): 弱酸洗脱肽负载的树突状细胞激活T细胞的实验研究

Arid

	弱酸洗脱肽负载的树突状细胞激活T细胞的实验研究
其他题名	Activation of T lymphocytes in vitro by dendritic cells loaded with mild acid-eluted peptides
	邹文燕 1; 徐军 1; 钱志红 2; 王治洁 1; 孙梅芳 1; 石礼红 1; 林晓 1
ISSN	0253-9934
出版年	2007
卷号	30 期号:5 页码:358-361
中文摘要	目的采用弱酸洗脱提取人卵巢癌细胞膜表面的肿瘤抗原肽,用于负载脐血树突状细胞(DCS),观察其对混合T淋巴细胞的体外激活效应.方法以淋巴细胞分离液分离脐血中的单个核细胞(CBMNC),将其中贴壁的细胞在细胞因子重组人粒巨噬细胞集落刺激因子(rhGM-CSF)、重组白介素(rhIL)-4及重组肿瘤坏死因子(rhTNF-alpha)的配伍作用下诱导分化为DCs.室温下以pH为3.3的枸橼酸一磷酸盐缓冲液处理人卵巢癌细胞,洗脱液经Oasis HLB小柱提取纯化后负载DCs.以~3H-TdR掺人法测定负载抗原的DCs对混合T淋巴细胞的体外活化效应.结果贴壁的CBMNC在细胞因子2000 U/mL rhGM-CSF、500U/mL rhIL-4及100 U/mL rhTNF-alpha的配伍作用下体外诱导分化为具有典型形态和表型的DCs.人卵巢癌细胞弱酸洗脱肽负载的DCs与混合T淋巴细胞体外共培养后,T细胞增殖加快.结论 CBMNC可以在GM-CSF、IL-4和TNF-alpha的作用下体外诱导分化为功能性DCs.pH为3.3的枸橼酸-磷酸盐缓冲液洗脱法可得到卵巢癌细胞的弱酸洗脱肽,用其负载脐血DCs,体外可激活细胞毒T淋巴细胞.
英文摘要	Objective To observe the in vitro activation effect of cord blood-derived dendritic cells (DCs) loaded with mild acid-eluted peptides of HO-8910 tumor cells on T lymphocyte. Methods Cord blood was collected from healthy term-delivery mother. The cord blood mononuclear cells(CBMNCs) were separated by density gradient centrifugation(Ficoll-Hypaque). The adherent fraction of CBMNCs was cultured in presence of relevant cyto-kines(rhGM-CSF 2 000 U/mL, rhIL-4 500 U/mL and rhTNF-alpha100 U/mL) to generate DCs. The tumor cells of human ovarian carcinoma cell line HO-8910 were treated with citrate-phosphate buffer(pH 3. 3) and the washing fluid was subjected to an Oasis HLB solid-phase extraction cartridge for mild acid-eluted peptides. As tumor antigens, mild-acid eluted peptides were used to prime the DCs in vitro. The stimulation ability of DCs on the T lymphocyte proliferation was reflected by mixed T lymphocyte reaction using 3H-TdR incorporation. Results DCs with typical morphology and phenotype were harvested from the adherent fraction of CBMNCs in presence of rhGM-CSF 2 000 U/mL, rhIL-4 500 U/mL and rhTNF-a 100 U/mL in vitro. Loaded with the mild acid-eluted peptides, DCs were able to stimulate the amplification of allogeneic mixed T lymphocytes. The stimulate index(SI) of the loaded groups were higher than the unloading group at an effector : target ratios of 1 : 10, 1 : 50 and 1: 100. Conclusion Functional DCs with typical morphology and phenotype had been successfully harvested from the adherent fraction of CBMNCs in presence of rhGM-CSF, rhIL-4 and rhTNF-alphain vitro. After loading with mild acide-luted peptides of ovarian carcinoma cells, the cord blood-derived DCs can activate CTL in vitro.
中文关键词	树突状细胞 ; 卵巢癌 ; 弱酸洗脱肽 ; 脐血
英文关键词	Dendritic cells Ovarian carcinoma Mild acid-eluted peptides Cord blood
语种	中文
国家	中国
收录类别	CSCD
WOS类目	PHARMACOLOGY PHARMACY
WOS研究方向	Pharmacology & Pharmacy
CSCD记录号	CSCD:2822121
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/221297
作者单位	1.上海市第八人民医院妇产科, 200235; 2.苏州大学附属二院妇产科
推荐引用方式 GB/T 7714	邹文燕,徐军,钱志红,等. 弱酸洗脱肽负载的树突状细胞激活T细胞的实验研究[J],2007,30(5):358-361.
APA	邹文燕.,徐军.,钱志红.,王治洁.,孙梅芳.,...&林晓.(2007).弱酸洗脱肽负载的树突状细胞激活T细胞的实验研究.,30(5),358-361.
MLA	邹文燕,et al."弱酸洗脱肽负载的树突状细胞激活T细胞的实验研究".30.5(2007):358-361.