抗人DC和抗大鼠DC单抗杂交瘤的细胞特异性鉴定

Primary identification of the anti-human dendritic cells and antirat dendritic cells monoclonal antibody by flow cytometer

目的:进一步鉴定在以往研究中所获得的抗人树突状细胞(dendritic cells,DCs)单抗和抗大鼠单抗。方法:通过对GM-CSF和IL-4诱导人外周血获得人的DC和各种来源的造血系统肿瘤细胞系的直接和间接荧光染色,采用流式细胞仪分析这些单抗所识别的表位在人DC上的表达情况以及与其他细胞表达表位的交叉情况。结果:D5B10所识别的表位仅与K562细胞有较大的交叉性,而与其他来源的白血病细胞均无明显的交叉。72%左右的人外周血成熟DC存在D5B10和H1E11识别的表位;WZD3识别的表位与人DC交叉较少,仅24.44%。三种单抗所识别的表位在MTDC上均有,大约有一半的MTDC表达三种单抗所识别的表位。结论:本实验室所制备的抗人DC单抗和抗大鼠DC单抗所识别的表位在人和小鼠间存在着交叉,这意味着这些单抗在小鼠DC的研究中也可使用。此外抗大鼠DC单抗所识别的表位与人DCs间存在着交叉。由于以往在制备抗人DCs单抗时是采用外周血分离的DCs,因此本次研究的结果也表明外周血分离的DC和由GM-CSF,IL-4诱导的DC可能在成熟程度和表达的各类分子水平存在区别。

Objective： We have established some hybridomas to anti-human dendritic cells（D5B10、H1E11） and anti-rat dendritic cells（WZD3） successfully.In order to use these monoclonal antibodies for the further research work we used the flow cytometer to identify them.Methods： Using flow cytometer we detected epitope on the human DCs（induced from human peripheral blood monocytes by GM-CSF and IL-4）,mouse thymic dendritic cell line（MTDC） and some blood cancer cell lines. Results： We found that about 50% K562 cells had the epitope identified by D5B10,but few on the other blood cancer cell lines.About 72% mature DCs from human peripheral monocytes were H1E11and D5B10 positive,the lowest positive were WZD3 only 24.44%,and about half MTDC were stained by these three monoclonal antibodies. Conclusion： The results expressed that there were some common epitopes between the human peripheral blood DCs and mouse MTDC.It also declared that DCs isolated human peripheral blood were different from DCs induced by GM-CSF and IL-4;for the preparation of D5B10 and H1E11 isolated human peripheral blood DCs were used.