蛛网膜细胞与血性CSF共同培养后HLA-DR的表达变化

The alteration of expression of HLA-DR after coculturing arachnoid cells in vitro with hematic CSF

目的研究蛛网膜细胞（Ar细胞）与血性脑脊液（CSF）共同培养后人类白细胞抗原- DR（HLA-DR）的表达变化。方法（1）Ar细胞的培养、鉴定和HLA-DR基础表达的观察。（2）采用Ar细胞和血性CSF共同培养的方法,用流式细胞仪（FCM）和免疫荧光染色观察Ar细胞HLA-DR的表达改变。结果95%以上培养的Ar细胞均有HLA-DR的基础表达。在血性CSF刺激下,Ar细胞HLA-DR的表达明显增强,免疫荧光法和FCM检测出了一致的时间表达变化模式,在1-5d呈基础表达状态;6d左右表达开始升高;7d、8d表达显著增强,此时荧光颗粒数量和强度最高,阳性细胞数可达61．2%;第9天后表达逐渐降低,阳性细胞数减少,第10天时减低至12．5%,荧光数量和强度亦明显降低。结论在血性CSF刺激下,Ar细胞通过HLA-DR表达增高,可以发挥抗体提呈细胞（APC）的功能,提示其可能与SAH并发症尤其是迟发性脑缺血（DCI）的发生机制相关。

Objective To study the alteration of the expression of HLA-DR after cocuhuring arachnoid cells in vitro with hematic cerebrospinal flood （CSF）. Methods （ 1 ） The arachnoid cells were cultured, characterized and examined by immunofluorescence for the basal expression of HLA-DR. （2）The alteration of the expression of HLA-DR after cocuhuring arachnoid cells in vitro with hematic cerebrospinal flood （CSF） was investigated by immunofluorescence and FACS. Results （1）Arachnoid cells were successfully cultured for many passages. Immunofluorescence stained the HLA-DR of over 95% of the human arachnoid cells. The punctate HLA-DR was distributed in cytoplasm and not in karyon. （2） After cocuhure arachnoid cells in vitro with hematic CSF, many particles with strong fluorescence appeared in cytoplasm on the 6th day. On the 8th day, the quantity of particles and their fluorescence intensity arrived at the vertex. FACS showed that the percentage of HLA-DR expressing cells maintained aronud 2.5% in the early 6 days and increased suddenly to 61.2% on the 7th day. The finding of HLA-DR expression of immumofluorescence and FCM were consistent on time mode. Conclusion After stimulated by hematic CSF arachnoid cells have the potential to serve as antigen presenting cells, which indicates that arachnoid cells are involved in the occurrence of DCI after SAH.