真核表达hCAP-18/LL-37成熟肽对树突状细胞表面分子表达的影响

J4 ›› 2009, Vol. 35 ›› Issue (3): 482-486.

真核表达hCAP-18/LL-37成熟肽对树突状细胞表面分子表达的影响

袁红艳¹, 王凤丽¹, 许娜^1|2, 常雅萍¹

1．吉林大学基础医学院免疫学教研室|吉林长春130021；2．吉林医药学院|吉林吉林 132013

收稿日期:2008-10-29 出版日期:2009-05-28 发布日期:2009-08-14
通讯作者: 常雅萍 E-mail:cyping823@163.com
作者简介:袁红艳（1976-）|女|吉林省桦甸市人|讲师|医学博士|主要从事细胞与分子免疫学研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题（200705357）

Effects of eukaryotic expressive mature peptide of hCAP-18/LL-37 on expressions of membrane molecules on dendritic cells

YUAN Gong-Yan¹, WANG Feng-Li¹, HU Na^1|2, CHANG Ya-Ping¹

1. Department of Immunology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;2.Jilin Medical College,Jilin 132013,China

Received:2008-10-29 Online:2009-05-28 Published:2009-08-14

摘要/Abstract

摘要：

目的：研究真核表达的人源阳离子抗菌肽-18(hCAP-18)成熟肽LL-37对树突状细胞(DCs)表面分子表达的影响。方法：通过基因克隆方法构建hCAP-18成熟肽LL-37真核表达载体pcDNA4/Myc-His-LL-37，转染人胚肾HEK293细胞株，转染48 h后收获培养上清，与体外以rh-GM-CSF、rh-IL-4联合培养定向诱导分化的不成熟DCs共培养48 h，收获DCs；流式细胞仪检测DCs表面分子CD40及HLA-DR的表达。结果：成功构建成熟肽真核表达载体pcDNA4/Myc-His-LL-37，并实现在其真核细胞HEK293中的表达；流式细胞仪结果显示，pDNA4/Myc-His-LL-37基因转染HEK293细胞培养上清刺激组DCs中CD40及HLA-DR的表达阳性率与对照组比较均明显升高(P<0.05)。结论：构建了真核表达的hCAP-18成熟肽LL-37，其可上调DCs表面分子CD40及HLA-DR的表达，促进DCs功能的成熟。

关键词: hCAP-18/LL-37；成熟肽；树突状细胞；表面分子

Abstract:

Abstract:Objective To study the effects of eukaryotic expressive mature peptide LL-37 of human cationic antimicrobial peptide (hCAP-18) on the expressions of membrane molecules on dendritic cells (DCs). Methods By gene cloning,the eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 for the mature peptide LL-37 of hCAP-18 was constructed.Then they were transfected into HEK293 cell lines.After the cell lines were cultivated for 48 h，the supernatant was collected.Then the DCs from peripheral blood mononuclear cells (PBMCs) induced by rh-GM-CSF and rh-IL-4 were cultivated with the supernatant for 48h.The expressions of membrane molecules CD40 and HLA-DR on DCs were detected by flow cytometry (FCM).
Results The eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 was constructed successfully and it expressed in eukaryotic cells HEK293.FCM results indicated that the expressions of CD40 and HLA-DR on the membrane of DCs which were stimulated by the supernatant produced by pcDNA4/Myc-His-LL-37 were higher than those in control group(P<0.05). Conclusion The mature peptide LL-37 of hCAP-18 is constructed successfully and it can increase the expressions of the membrane molecules on DCs such as CD40 and HLA-DR.

Key words: hCAP-18/LL-37;mature peptide;dendritic cells;membrane molecules

中图分类号:

Q516

袁红艳, 王凤丽, 许娜, 常雅萍. 真核表达hCAP-18/LL-37成熟肽对树突状细胞表面分子表达的影响[J]. J4, 2009, 35(3): 482-486.

YUAN Gong-Yan, WANG Feng-Li, HU Na, CHANG Ya-Ping. Effects of eukaryotic expressive mature peptide of hCAP-18/LL-37 on expressions of membrane molecules on dendritic cells[J]. J4, 2009, 35(3): 482-486.