SARS肺间质血管病变的病理观察

目的 观察分析严重急性呼吸综合征(severe acute respiratory syndrome，SARS)肺间质血管病变的病理形态学特征。方法 对3例行尸体解剖的SARS病例肺组织进行常规HE染色、组织化学染色和免疫组织化学染色，并探讨其病理形态学特征。结果 死亡的SARS病人肺部病变除弥漫性的肺部损伤(肺泡腔内可见细颗粒样或泡状水肿液、脱落的肺泡上皮细胞；肺泡间隔内表面可见透明膜覆盖，有的区域肺泡腔内出现机化性改变等)外，肺间质内的毛细血管簇状增生及血管壁出现明显的损伤性改变。结论 SARS的肺部血管病变是一个特征性病理形态学变化，可能与SARS病毒的蛋白导致的剧烈免疫反应有关。     

SARS尸检组织的病理变化和超微结构观察

目的 研究严重急性呼吸综合征(SARS)尸检组织的临床病理和超微结构特征。方法 对1例SARS死亡患者做即刻肺穿刺和12h后尸检,进行病理形态和超微结构的观察;用Macchiavello法做病毒包涵体染色;并对淋巴结、脾脏、结肠、小肠及骨髓组织行CD20、CD45RO(UCHL-1)、CD4、CD8、CD68、CD34免疫组织化学标记。结果 SARS肺的主要病变为急性弥漫性全小叶性间质性炎,可见肺泡腔内透明膜形成和增生及脱落的肺泡上皮,偶见胞质内病毒包涵体样结构,病毒包涵体染色阳性,肺内小血管增生、扩张,呈血管炎性改变。淋巴结、脾脏结构破坏,淋巴滤泡消失,脾小体萎缩,淋巴细胞明显减少,组织细胞增生;结肠、小肠孤立和集合淋巴结淋巴滤泡消失;骨髓增生减低,巨核细胞增多。免疫组织化学染色：淋巴结、脾脏B细胞CD20弥漫散在阳性,CD45RO(UCHL-1)散在阳性,CD4辅助T细胞显著减少,CD8毒性T细胞稍增加,CD4／CD8比例明显小于0．5。电镜观察：肺泡内的单核巨噬细胞、肺泡上皮胞质内可见病毒样颗粒,大小80～160nm,有光晕或花环状包膜。结论 肺部明显急性弥漫性全小叶性间质性炎,肺泡腔透明膜形成,肺外淋巴造血系统明显损害,尤T细胞明显;内脏器官出血、坏死和血管炎改变等为急性SARS的形态特征;肺内所见病毒样颗粒可能为新型冠状病毒,推测其为此次SARS流行的主要病原体。     

肺泡蛋白沉积症的病理学特点与诊断

目的探讨肺泡蛋白沉积症（PAP）的病理学特点及其诊断方法。方法对39例PAP进行了常规的光镜观察,并用淀粉酶消化后过碘酸雪夫染色（D—PAS）、淀粉酶消化后黏液卡红（D-黏卡）、奥新兰（AB）进行组织化学染色。结果肺活检及尸检肺组织的病理改变有如下特点：（1）肺泡腔内及部分小支气管腔内充满嗜伊红性细颗粒状蛋白性物质,并有针状裂隙。（2）蛋白性物质中杂有多少不等的退变的肺泡上皮细胞及脱落的肺泡上皮细胞。（3）肺泡Ⅱ型上皮细胞增生。（4）肺泡间隔毛细血管充血,间质炎症不明显。（5）支气管壁炎症不明显。（6）周围肺组织代偿性肺气肿。支气管肺泡灌洗液（BALF）石蜡包埋切片显微镜下可见片状嗜伊红性细颗粒状蛋白性物质,其中可见针状裂隙。组织化学染色结果肺泡腔内或BALF中嗜伊红性细颗粒状蛋白性物质D—PAS阳性（呈红色）,D-黏卡阴性,AB弱阳性（淡蓝色）或阴性。肺活检及BALF取得标本是诊断PAP的重要方法和途径。结论典型的PAP肺泡腔内或BALF中出现均质嗜伊红性细颗粒状蛋白性物质并有针状裂隙,蛋白性物质D—PAS阳性,D-黏卡阴性具有诊断意义。     

SARS肺的病理鉴别诊断

目的 分析严重急性呼吸综合征(severe acute respiratory syndrome，SARS)肺部病变的临床病理形态学特征及与其他肺部炎性疾病的鉴别诊断。方法 对3例行尸体解剖的SARS病例的肺组织进行组织形态学、免疫组织化学和电镜下超微结构的观察，对其临床表现、病理形态学特征及主要鉴别诊断进行探讨，并与病理资料完整的16例病毒性肺炎、13例间质性肺炎的肺部病变进行鉴别诊断。结果 死亡的SARS病人肺部病变主要表现为弥漫性的肺部损伤，但是各处病变轻重不一。肺泡腔内可见细颗粒样或泡状水肿液，其中可见脱落的肺泡上皮细胞。肺泡间隔表面可见透明膜覆盖，有的区域肺泡腔内出现机化性改变，肺泡腔和肺问质内可见大量的巨噬细胞浸润。同时，肺间质内的小动脉壁出现明显的损伤性改变。结论 SARS的肺部病理形态学特征可与其他肺部炎症性疾病鉴别。     

严重急性呼吸综合征肺部病变发展过程的形态观察

目的探讨严重急性呼吸综合征(SARS)肺部病变发展过程中组织形态和免疫表型的变化.方法对4例SARS的肺标本进行病理形态和1例超微结构观察,并作VG、Masson、网状纤维、地依红、PAS和天狼猩红组织化学染色及采用链霉素抗生物素蛋白-过氧化物酶(SP)法作结蛋白、波形蛋白、细胞角蛋白(CK)、上皮膜抗原(EMA)、平滑肌肌动蛋白(SMA)、HHF-35、CD34、CD68、bel-2、第八因子相关抗原(FⅧRAg)、AE1、Ⅰ型胶原、Ⅲ型胶原免疫组织化学标记.结果依病程及病变损害程度分为3期.急性渗出期2例病程＜20 d.以急性渗出性肺泡炎、肺泡上皮细胞增生,坏变和肺透明膜形成、脱屑性肺炎为主要病变,局部有纤维母细胞增生;肺泡内渗出蛋白PAS阳性,网状纤维增多并断裂,纤维母细胞明显增多,结蛋白、波形蛋白阳性.纤维增殖期1例病程25 d.以增殖性间质肺炎为主,早期纤维化表现为机化性肺炎和血管周围机化性改变.网状纤维呈肾小球样增生,间叶细胞向肌纤维母细胞分化,结蛋白、HHF-35、SMA、波形蛋白呈强阳性;向平滑肌细胞分化结蛋白、SMA呈阳性,波形蛋白阴性;向纤维母细胞分化仅波形蛋白阳性,并见弹力纤维增多,断裂,胶原纤维增生.纤维化期1例75 d.以弥漫性纤维化,以纤维增生为主的伴蜂窝肺形成.天狼猩红染色胶原纤维显红色,波形蛋白和SMA弱阳性,Ⅰ、Ⅳ胶原纤维明显增多.各期巨噬细胞均增多,CD68阳性.结论SARS肺部病变可分为3期,随病程延长,肺纤维化进行性加重;肺泡壁原始间叶细胞,肺泡上皮细胞增生、损伤和增生的巨噬细胞在肺纤维化中起着重要作用.     

乳腺黏液囊肿样病变临床病理特征分析

目的探讨乳腺黏液囊肿样病变的临床病理特征及其诊断和鉴别诊断。方法对9例乳腺黏液囊肿样病变进行临床病理分析。以SP法对细胞角蛋白AE1／AE3、平滑肌肌动蛋白（SMA）、p63、c-erbB-2和053进行免疫组织化学染色,组织化学为AB和PAS法染色。结果患者均为女性,年龄23-43岁（平均34岁）,乳腺均可触及肿物。肿物切面呈多囊性胶冻样。镜下可见多发性、充满黏液的囊肿,囊腔衬覆扁平-立方-柱状上皮,其中3例上皮有乳头状增生,1例有不典型增生。间质内可见黏液湖,其内无漂浮细胞。囊内及间质黏液呈AB及PAS染色阳性。囊肿衬覆上皮细胞及2例黏液湖内漂浮的上皮细胞团AE1／AE3阳性。囊肿上皮细胞外侧扁平肌上皮呈SMA、p63阳性。c-erbB-2和p53染色均阴性。结论乳腺黏液囊肿样病变是一种独立的良性病变,容易误诊,需与黏液癌等肿瘤进行鉴别。     

严重急性呼吸综合征病原体检测及临床病理学观察

目的 研究重症急性呼吸综合征(SARS)的临床病理学特点。方法 利用3例尸检材料为观察对象,全部材料经常规HE染色,肺组织部分标本经组织化学Macchiavello法(病毒包涵体染色)、网状纤维、PAS染色;免疫组织化学;超薄切片及染色;光镜及透射电镜观察。结果 3例均以高热为首发症状,继而出现进行性呼吸困难和肺部阴影;肺部病变：双肺广泛性实变;灶性出血,坏死,脱屑性肺泡炎及支气管炎,肺泡腔内充满增生脱落的肺泡上皮及渗出的蛋白、单核细胞、淋巴细胞和浆细胞,肺透明膜形成,部分肺泡腔内、渗出物机化呈肾小球样机化性肺炎改变,肺泡上皮细胞内可见病毒包涵体;免疫器官损伤：脾脏淋巴组织大片状坏死,淋巴结灶性坏死。骨髓造血组织抑制;全身小血管炎：心、肺、肝、肾、肾上腺、横纹肌间小静脉周围及血管壁水肿,灶性纤维素样坏死,单核细胞、淋巴细胞浸润,部分小静脉有血栓形成;全身中毒性改变：肺、肝、肾、心、肾上腺实质细胞变性,坏死。肺组织透射电镜观察发现群集的病毒颗粒。结论本病是一种全身多器官损伤性疾病,肺部及免疫系统是病毒主要作用的靶器官,肺部广泛性实变,大量透明膜形成,呼吸窘迫及免疫功能低下是本病死亡的主要原因。     

肺上皮样血管内皮瘤临床病理观察

目的 探讨肺上皮样血管内皮瘤的临床病理特点。方法 4例肺上皮样血管内皮瘤,3例女性,1例男性,年龄28～40岁,无自觉症状或有轻度咳嗽、气短。肺活检或手术切除标本经甲醛固定,石蜡包埋,常规HE及免疫组织化学(Envision法)染色。所用抗体包括CD31、CD34、细胞角蛋白(AEl／AE3)、TTF-1、波形蛋白和上皮膜抗原。结果 本组肺上皮样血管内皮瘤病例女性多于男性,胸部CT显示双肺多发弥漫性小结节影。病理形态特点为结节周边上皮样肿瘤细胞呈花冠状充填于肺泡腔,病变中心为黏液透明样变间质,肺泡壁结构保留,肿瘤细胞胞质内有空泡形成,空泡内偶见红细胞,肿瘤细胞异型性不明显,核分裂和坏死均少见,免疫组织化学染色示CD31、CD34阳性,AE1／AE3偶见灶状阳性,其他抗体呈阴性。结论 肺上皮样血管内皮瘤是一种具有独特临床病理特点的低度恶性血管来源肿瘤。     

肺黏膜相关淋巴组织边缘区B细胞淋巴瘤及良性淋巴组织增生性疾病的临床病理分析

目的 研究肺原发性黏膜相关淋巴组织边缘区B细胞(MALT)淋巴瘤及良性淋巴组织增生性疾病的临床病理形态、免疫组织化学表型和B细胞重链基因重排,比较肺MALT淋巴瘤和良性淋巴组织增生性疾病的差异.方法 回顾性的分析原发性肺MALT淋巴瘤13例,7例肺良性淋巴组织增生性疾病资料.对标本行常规HE染色,EnVision免疫组织化学染色(抗体包括AE1/AE3、CD20、CD79α、CD3、CD5、CD10、CD21、bel-2、bcl-6、cyclinD-1)及免疫球蛋白重链IgH基因重排检测.结果 13例肺MALT淋巴瘤,细胞成分多样,分别由不同比例的小淋巴细胞样细胞、中心细胞样细胞、单核样B细胞组成,常伴有浆细胞分化.肿瘤细胞以弥漫性和滤泡边缘区排列为主,常见反应性淋巴滤泡和滤泡中心的植入.肿瘤细胞呈串珠状直接侵犯肺泡间隔和沿支气管血管束向周边及肺膜扩散.MALT淋巴瘤中,均未见坏死.9例可见肿瘤细胞侵犯血管壁,6例可见胸膜累及,2例肺门淋巴结侵犯.9例肺MALT淋巴瘤可见淋巴上皮样病变,免疫组织化学显示上皮细胞内的淋巴细胞CD20阳性,CD3阴性.7例肺良性淋巴组织增生性疾病,2例可见淋巴上皮样病变,免疫组织化学显示,其淋巴上皮样病变内的淋巴细胞,部分CD20阳性,部分CD3阳性.9例肺MALT淋巴瘤进行了免疫球蛋白重链IgH基因重排,8例阳性;7例良性淋巴组织增生性疾病均为阴性.结论 肺MALT淋巴瘤在细胞组成和排列上与其他部位结外MALT淋巴瘤相同,肿瘤细胞呈串珠状直接侵犯肺泡间隔和沿支气管血管束向周边及肺膜扩散.在肺内淋巴上皮样病变常见于MALT淋巴瘤,并有助于诊断,但并非其特异性病变,一些肺的反应性淋巴组织增生也可出现,用免疫组织化学有助于区别两种病变.免疫球蛋白重链IgH基因重排可以帮助鉴别肺MALT淋巴瘤和良性淋巴组织增生性疾病.     

严重急性呼吸综合征临床病理分析

目的 探讨严重急性呼吸综合征(Severe acute respitatory syndrome，SARS)的病理形态学特点。方法 3例SARS标本为患者支气管镜穿刺活检和切取的肺组织，经HE染色，光镜观察，并用免疫组化S-P法检测SARS患者肺组织CK、CD34、Ⅳ型胶原、CD68、Mac387的表达。结果 SARS患者肺组织病理形态表现为肺泡上皮广泛破坏，Ⅱ型上皮细胞增生。肺泡壁毛细血管内皮细胞肿胀。大部分肺泡腔及肺泡管内透明膜形成。间质有较多的炎细胞浸润，成纤维细胞增生。结论 SARS是一种伴透明膜形成的急性非特异性间质性肺炎。     

Immunohistochemical, in situ hybridization, and ultrastructural localization of SARS-associated coronavirus in lung of a fatal case of severe acute respiratory syndrome in Taiwan

This article describes the pathological studies of fatal severe acute respiratory syndrome (SARS) in a 73-year-old man during an outbreak of SARS in Taiwan, 2003. Eight days before onset of symptoms, he visited a municipal hospital that was later identified as the epicenter of a large outbreak of SARS. On admission to National Taiwan University Hospital in Taipei, the patient experienced chest tightness, progressive dyspnea, and low-grade fever. His condition rapidly deteriorated with increasing respiratory difficulty, and he died 7 days after admission. The most prominent histopathologic finding was diffuse alveolar damage of the lung. Immunohistochemical and in situ hybridization assays demonstrated evidence of SARS-associated coronavirus (SARS-CoV) infection in various respiratory epithelial cells, predominantly type II pneumocytes, and in alveolar macrophages in the lung. Electron microscopic examination also revealed coronavirus particles in the pneumocytes, and their identity was confirmed as SARS-CoV by immunogold labeling electron microscopy. This report is the first to describe the cellular localization of SARS-CoV in human lung tissue by using a combination of immunohistochemistry, double-stain immunohistochemistry, in situ hybridization, electron microscopy, and immunogold labeling electron microscopy. These techniques represent valuable laboratory diagnostic modalities and provide insights into the pathogenesis of this emerging infection.     

Pulmonary pathological features in coronavirus associated severe acute respiratory syndrome (SARS)

BACKGROUND: Severe acute respiratory syndrome (SARS) became a worldwide outbreak with a mortality of 9.2%. This new human emergent infectious disease is dominated by severe lower respiratory illness and is aetiologically linked to a new coronavirus (SARS-CoV). METHODS: Pulmonary pathology and clinical correlates were investigated in seven patients who died of SARS in whom there was a strong epidemiological link. Investigations include a review of clinical features, morphological assessment, histochemical and immunohistochemical stainings, ultrastructural study, and virological investigations in postmortem tissue. RESULTS: Positive viral culture for coronavirus was detected in most premortem nasopharyngeal aspirate specimens (five of six) and postmortem lung tissues (two of seven). Viral particles, consistent with coronavirus, could be detected in lung pneumocytes in most of the patients. These features suggested that pneumocytes are probably the primary target of infection. The pathological features were dominated by diffuse alveolar damage, with the presence of multinucleated pneumocytes. Fibrogranulation tissue proliferation in small airways and airspaces (bronchiolitis obliterans organising pneumonia-like lesions) in subpleural locations was also seen in some patients. CONCLUSIONS: Viable SARS-CoV could be isolated from postmortem tissues. Postmortem examination allows tissue to be sampled for virological investigations and ultrastructural examination, and when coupled with the appropriate lung morphological changes, is valuable to confirm the diagnosis of SARS-CoV, particularly in clinically unapparent or suspicious but unconfirmed cases.     

SARS患者组织中冠状病毒S蛋白的表达

目的探讨冠状病毒S蛋白单克隆抗体在5例严重急性呼吸综合征(severeacuterespiratorysyndrome,SARS)患者多脏器穿刺和尸检病理组织中的表达以及与SARS病理学特点及病原学的关系。方法应用SARS冠状病毒(SARSCoV)S蛋白单克隆抗体,对病程为1周的早期SARS死亡尸检病例和4例病程为3～5周中晚期SARS死亡病例的多器官组织进行检测和观察。结果肺泡腔内脱落的上皮细胞、单核细胞和多核合体样细胞,以及支气管黏膜上皮细胞与黏膜腺体上皮细胞、多器官内血管内皮细胞和散在浸润的单核巨噬细胞SARSCoVS蛋白均呈阳性表达。2例SARS中晚期病例肝细胞和心肌细胞亦见SARSCoVS蛋白阳性反应。结论SARSCoV单克隆抗体在SARS患者肺、支气管、肝脏和心肌等器官组织均有表达,可作为SARS病原学和病理学的辅助诊断标志,对探讨SARS传播途径及发病机制有重要意义。     

Tissue and cellular tropism of the coronavirus associated with severe acute respiratory syndrome: an in-situ hybridization study of fatal cases

Severe acute respiratory syndrome (SARS) is a new human infectious disease with significant morbidity and mortality. The disease has been shown to be associated with a new coronavirus (SARS-CoV). The clinical and epidemiological aspects of SARS have been described. Moreover, the viral genome of SARS-CoV has been fully sequenced. However, much of the biological behaviour of the virus is not known and data on the tissue and cellular tropism of SARS-CoV are limited. In this study, six fatal cases of SARS were investigated for the tissue and cellular tropism of SARS-CoV using an in-situ hybridization (ISH) technique. Among all the tissues studied, positive signals were seen in pneumocytes in the lungs and surface enterocytes in the small bowel. Infected pneumocytes were further confirmed by immunofluorescence-fluorescence in-situ hybridization (FISH) analysis. These results provide important information concerning the tissue tropism of SARS-CoV, which is distinct from previously identified human coronaviruses, and suggest the possible involvement of novel receptors in this infection. Whereas the lung pathology was dominated by diffuse alveolar damage, the gut was relatively intact. These findings indicated that tissue responses to SARS-CoV infection are distinct in different organs.     

Lung pathology of severe acute respiratory syndrome (SARS): a study of 8 autopsy cases from Singapore

Severe acute respiratory syndrome (SARS) is an infectious condition caused by the SARS-associated coronavirus (SARS-CoV), a new member in the family Coronaviridae. To evaluate the lung pathology in this life-threatening respiratory illness, we studied postmortem lung sections from 8 patients who died from SARS during the spring 2003 Singapore outbreak. The predominant pattern of lung injury in all 8 cases was diffuse alveolar damage. The histology varied according to duration of illness. Cases that were 10 or fewer days in duration demonstrated acute phase diffuse alveolar damage (DAD), airspace edema, and bronchiolar fibrin. Cases with a time course greater than 10 days showed organizing phase DAD, type II pneumocyte hyperplasia, squamous metaplasia, multinucleated giant cells, and acute bronchopneumonia. In acute DAD, pancytokeratin staining was positive in hyaline membranes along alveolar walls and highlighted the absence of pneumocytes. Multinucleated cells were shown to be both type II pneumocytes and macrophages by pancytokeratin, TTF-1, and CD68 staining. SARS-CoV RNA was identified by RT-PCR in 7 of 8 cases in fresh autopsy tissue and in 8 of 8 cases in formalin-fixed, paraffin-embedded lung tissue including the one negative case in fresh tissue. Understanding the pathology of DAD in SARS patients may provide the basis for therapeutic strategies. Further studies of the pathogenesis of SARS may reveal new insights into mechanisms of DAD.     

原位RT-PCR法测定SARS患者肺组织中的病毒核酸

【摘要】目的观察严重急性呼吸综合征(SARS)患者肺组织中SARS病毒核酸的定位和分布。方法采用原位逆转录一聚合酶链反应(RT-PCR)技术对3例SARS死亡患者肺组织进行病毒核酸测定。以固定在载玻片上的肺组织为靶细胞，直接掺入地高辛素一dN31P标记物进行PCR扩增，并观察病毒核酸阳性物质。结果检测3例尸解肺组织，其中2例可见SARS病毒核酸阳性信号。阳性物质表现为蓝紫色颗粒状物，分布于细胞核或细胞浆内。在肺泡上皮细胞、支气管上皮细胞及单个核细胞中均可观察到该病毒核酸物质。肺组织病理改变明显，表现为出血、渗出，炎性浸润等改变。结论在肺组织中检出SARS病毒核酸物质，并与病理改变并存，为本病的病原学诊断提供直接证据。【关键词】严重急性呼吸综合征；SARS病毒；原位逆转录一聚合酶链反应；肺组织     

Pathology and virus dispersion in cynomolgus monkeys experimentally infected with severe acute respiratory syndrome coronavirus via different inoculation routes

Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) causes SARS. The pathogenic mechanisms of SARS-CoV remain poorly understood. Six cynomolgus monkeys were inoculated with the HKU39849 isolate of SARS-CoV via four routes. After intranasal inoculation, the virus was isolated from respiratory swabs on days 2-7 postinoculation (p.i.) and virus genome was detected in intestinal tissues on day 7 p.i. Virus was not detected after intragastric inoculation. After intravenous inoculation, infectious virus was isolated from rectal swabs, and virus antigen was detected in intestinal cells on day 14 p.i. After intratracheal (i.t.) inoculation, virus antigen-positive alveolar cells and macrophages were found in lung and infectious virus was detected in lymphoid and intestinal tissues. The peribronchial lymph nodes showed evidence of an immune response. Lung tissue and/or fluid and/or the peribronchial lymph node of the intratracheally inoculated animals had high TNF-alpha, IL-8 and IL-12 levels. SARS lung lesions are only generated in monkeys by i.t. inoculation. The virus appears to spread into and perhaps via the intestinal and lymphatic systems. It has been suggested previously that viraemia may cause intestinal infections in SARS patients.     

Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis

Severe acute respiratory syndrome (SARS) is an acute infectious disease that spreads mainly via the respiratory route. A distinct coronavirus (SARS-CoV) has been identified as the aetiological agent of SARS. Recently, a metallopeptidase named angiotensin-converting enzyme 2 (ACE2) has been identified as the functional receptor for SARS-CoV. Although ACE2 mRNA is known to be present in virtually all organs, its protein expression is largely unknown. Since identifying the possible route of infection has major implications for understanding the pathogenesis and future treatment strategies for SARS, the present study investigated the localization of ACE2 protein in various human organs (oral and nasal mucosa, nasopharynx, lung, stomach, small intestine, colon, skin, lymph nodes, thymus, bone marrow, spleen, liver, kidney, and brain). The most remarkable finding was the surface expression of ACE2 protein on lung alveolar epithelial cells and enterocytes of the small intestine. Furthermore, ACE2 was present in arterial and venous endothelial cells and arterial smooth muscle cells in all organs studied. In conclusion, ACE2 is abundantly present in humans in the epithelia of the lung and small intestine, which might provide possible routes of entry for the SARS-CoV. This epithelial expression, together with the presence of ACE2 in vascular endothelium, also provides a first step in understanding the pathogenesis of the main SARS disease manifestations.     

Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV)

Inactivated severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been tested as a candidate vaccine against the re-emergence of SARS. In order to understand the efficacy and safety of this approach, it is important to know the antibody specificities generated with inactivated SARS-CoV. In the current study, a panel of twelve monoclonal antibodies (mAbs) was established by immunizing Balb/c mice with the inactivated BJ01 strain of SARS-CoV isolated from the lung tissue of a SARS-infected Chinese patient. These mAbs could recognize SARS-CoV-infected cells by immunofluorescence analysis (IFA). Seven of them were mapped to the specific segments of recombinant spike (S) protein: six on S1 subunit (aa 12-798) and one on S2 subunit (aa 797-1192). High neutralizing titers against SARS-CoV were detected with two mAbs (1A5 and 2C5) targeting at a subdomain of S protein (aa 310-535), consistent with the previous report that this segment of S protein contains the major neutralizing domain. Some of these S-specific mAbs were able to recognize cleaved products of S protein in SARS-CoV-infected Vero E6 cells. None of the remaining five mAbs could recognize either of the recombinant S, N, M, or E antigens by ELISA. This study demonstrated that the inactivated SARS-CoV was able to preserve the immunogenicity of S protein including its major neutralizing domain. The relative ease with which these mAbs were generated against SARS-CoV virions further supports that subunit vaccination with S constructs may also be able to protect animals and perhaps humans. It is somewhat unexpected that no N-specific mAbs were identified albeit anti-N IgG was easily identified in SARS-CoV-infected patients. The availability of this panel of mAbs also provided potentially useful agents with applications in therapy, diagnosis, and basic research of SARS-CoV.