摘要
目的:了解肺组织内被严重急性呼吸综合征冠状病毒(severeacutere spiratorysyndromeas sociatedcoron avirus,SARS CoV)感染的靶细胞类型,并对SARS诱发的肺损伤发病机制进行探讨。方法:运用SARSCoV基因组序列合成的地高辛标记cDNA探针,对北京市7例及安徽省1例确诊的SARS死亡病例的肺组织进行原位杂交检测,在原位杂交基础上,进一步运用免疫组织化学方法显示SARSCoV感染的靶细胞类型,如Cytokeratin(CK)标记上皮细胞,CD34标记血管内皮细胞,CD68标记巨噬细胞,Vimentin标记纤维母细胞,CD3标记全T细胞。结果:原位杂交检测显示,8例患者肺组织中都表达SARSCoVRNA,阳性信号位于靶细胞胞浆内,呈紫蓝色(NBTBCIP)。原位杂交和免疫组化结果显示:支气管上皮细胞、Ⅱ型肺泡上皮细胞、血管内皮细胞、巨噬细胞、纤维母细胞及T淋巴细胞在所有SARS病例中都受到了病毒感染。原位杂交阳性(紫蓝色,NBTBCIP)和免疫组化阳性(红棕色,AEC)信号同时表达于靶细胞胞浆中而呈紫红色。结论:通过对SARS患者肺组织原位杂交和免疫组织化学双重标记研究表明,肺组织内支气管上皮细胞、Ⅱ型肺泡上皮细胞、血管内皮细胞、巨噬细胞及纤维母细胞等多种细胞成分广泛受到了SARSCoV攻击,肺组织内多种细胞成分弥漫性受损以及所释放的炎性介质在肺损伤的发病过程中起着重要作用。
Objective： To investigate the cell types infected by severe acute respiratory syndrome -associated coronavirus （SARS-CoV） in lung tissues and explore the machanism of lung injury in SARS. Methods： In-situ hybridization （ ISH ） and immunohistochemistry （IHC） double staining was applied to study the lung tissues.from 7 SARS cases of Beijing and one of Anhui province. According to SARS-CoV genome sequence, the cDNA probe was synthesized and labelled by digoxin. Immunohistochemically, antibodies of cytokeratin（CK） , CD34, CD68, Vimentin and CD3 were applied to demonstrate bronchial epithelial cells, type Ⅱ pnumocytes, endothelial cells, macrophages, fibroblasts and T cells respectively. Results： The positive results of in-situ hybridization showed that the lung tissues of all cases expressed SARS-CoV RNA, and positive signals displayed in cytoplasms （purple-blue, NBP-BCIP. ISH-IHC double staining showed that positive signals of both ISH （purple-blue NBT-BCIP and IHC （redbrown, AEC expressed in the cytoplasms（ purple and red）. The positive results of double staining indica- ted that bronchial epithelial cells, type Ⅱ pnumocytes, endothelial cells, macrophages,fibroblasts and T lymphocytes were diffusely infected by SARS-CoV. Conclusion： This study of ISH-IHC double staining in lung tissues of SARS patients showed that bronchial epithelial cells, type Ⅱ pnumocytes, endothelial cells, macrophages, T lymphocytes and fibroblasts were attacked diffusely in SARS lungs. Various types of cells damaged by SARS-CoV and inflammatory mediators released by those cells play an important role in the pathogenesis of lung injury in SARS.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2005年第5期453-457,共5页
Journal of Peking University:Health Sciences
基金
国家高技术研究发展计划专项经费(2003AA208105)
中华人民共和国科技部SARS专项基金(2003AA208107)资助~~
