[Expression of SARS-CoV in various types of cells in lung tissues].
OBJECTIVE
To investigate the cell types infected by severe acute respiratory syndrome-associated coronavirus (SARS-CoV) in lung tissues and explore the mechanism 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 pneumocytes, 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 (red-brown, AEC expressed in the cytoplasms (purple and red). The positive results of double staining indicated that bronchial epithelial cells, type II pneumocytes, endothelial cells, macrophages, fibroblasts and T lymphocytes were diffusely infected by SARS-CoV.


CONCLUSIONS
This study of ISH-IHC double staining in lung tissues of SARS patients showed that bronchial epithelial cells, type II pneumocytes, 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.
Lu
Yuan
Y
Department of Pathology, Peking University School of Basic Medical Sciences, Beijing 100083, China.
Gong
En-cong
EC
Zhang
Qian-ying
QY
Gu
Jiang
J
Li
Xing-wang
XW
Zhang
Bo
B
Hou
Lin
L
Shao
Hong-quan
HQ
Gao
Zi-fen
ZF
Zheng
Jie
J
Fang
Wei-gang
WG
Zhong
Yan-feng
YF
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D017252:CD3 Complex / Q000032:analysis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007633:Keratins / Q000032:analysis; D008168:Lung / Q000737:chemistry / Q000473:pathology / Q000821:virology*; D016676:Macrophages, Alveolar / Q000737:chemistry / Q000473:pathology / Q000821:virology; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000032:analysis; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D045169:Severe Acute Respiratory Syndrome / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D013601:T-Lymphocytes / Q000737:chemistry / Q000473:pathology / Q000821:virology; D014746:Vimentin / Q000032:analysis; D000097382:CD68 Molecule
D015703:Antigens, CD; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D017252:CD3 Complex; D007633:Keratins; D012367:RNA, Viral; D014746:Vimentin; C067980:CD68 antigen, human; D000097382:CD68 Molecule