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,pcDNA4/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。
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 HI.A-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 1.L-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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2009年第3期482-486,共5页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科技发展计划项目资助课题(200705357)
