Shi Yu-ling, Li Lin-hai, Sun Zhao-hui, Chen Jian-yun, Liao Yang, Zeng Lan-lan, Zhang Wei, Chen Xiao-dong, Cao Cheng
Clinic Laboratory, General Hospital of Guangzhou Command, Guangzhou, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2010 Jul;31(7):795-9.
To study the change of special antibodies titer IgG, IgM and nucleocapsid to SARS coronavirus (CoV) and observing the expression of stomach and enteric involvement on SARS-CoV infection by monoclonal antibody against N protein of SARS-CoV in the 7-year recovery period among family clustering cases of severe acute respiratory syndrome.
Special antibody titer to SARS-CoV of 14 patients from 5 different families and their 10 kinfolks continuously tested by IFA and antigen-capturing ELISA methods. Samples were taken in the 1(st) - 7(th) year periods after SARS patients infected by SARS-CoV, being diluted and measured on it titers of three kinds of antibodies. Immunochemical staining with monoclonal antibody (mAb) against N protein of SARS-CoV was used to determine the stomach and enteric tissues among 5 SARS patients with their nucleocapsid antibody titer ascended obviously after 1(st)-7(th) year.
When testing the IgG antibody titer of the 14 SARS patients by IFA method, the average titer was 1/71 (95%CI: 1/58 - 1/85) in the 1(st) year, but began to descend in the following years, and the IgG antibody of the most SARS patients disappeared in the 7(th) year. Regarding the IgM titer, it disappeared in most of the SARS patients 1 year later. The average value of nucleocapsid antibody titer was 1/146 (95%CI: 1/122 - 1/171) in the 1(st) year, and it descended as the IgG antibody titer did. In 5 cases, differences appeared. The nucleocapsid antibody titer was between 1/156 and 1/210 in 3 cases, and 2 cases were normal. Immunochemical staining with mAb against N protein of SARS-CoV was identified in the stomach and enteric tissues of 5 SARS patients with the nucleocapsid antibody titer increased significantly, 1(st)-7(th) year later. The five patients were detected by gastroscopy detection and cell immunohistochemistry test. 3 cases showed N protein antibody positive in the serum, and positive immunohistochemical expression in most of the cytoplasm in the gastric tissue mucous gland epithelial cells. 1 case also expressed in the intestinal tissue slurry columnar epithelium and interstitial cells. The other two cases showed negative on both serum N protein antibody and immunohistochemical expression. The biopsy results of the 5 patients were as follows: 1 case diagnosed as "signet-ring cell carcinoma of the stomach and rectum multiple transfer", 1 case of gastric polyp, 1 case of superficial antral gastritis and 2 cases were normal.
By testing the special IgG, IgM, nucleocapsid antibody to SARS-CoV of the 14 family clustering cases, we found that they all decreased in the 7(th) year, and most of them disappeared. The nucleocapsid antibody titer was related to pathogenetic condition. SARS-CoV was proved to be still present in stomach and enteric tissues of SARS patients with the nucleocapsid antibody titer increased significantly after the 7(th) year.
研究严重急性呼吸综合征家庭聚集性病例7年恢复期患者体内针对严重急性呼吸综合征冠状病毒(SARS-CoV)的特异性抗体IgG、IgM及核衣壳抗体滴度的变化,并通过抗SARS-CoV N蛋白单克隆抗体观察SARS-CoV感染时胃肠道受累情况。
采用间接免疫荧光法(IFA)和抗原捕获酶联免疫吸附试验(ELISA)连续检测来自5个不同家庭的14例患者及其10名亲属血清中SARS-CoV特异性抗体滴度。在SARS患者感染SARS-CoV后的第1至7年采集样本,稀释后检测3种抗体的滴度。采用抗SARS-CoV N蛋白单克隆抗体进行免疫组化染色,对核衣壳抗体滴度在第1至7年明显升高的5例SARS患者的胃肠道组织进行检测。
采用IFA法检测14例SARS患者IgG抗体滴度,第1年平均滴度为1/71(95%可信区间:1/58 - 1/85),随后逐年下降,多数患者的IgG抗体在第7年消失。IgM抗体滴度方面,多数SARS患者1年后消失。核衣壳抗体滴度第1年平均值为1/146(95%可信区间:1/122 - 1/171),与IgG抗体滴度一样下降。5例出现差异,3例核衣壳抗体滴度在1/156至1/210之间,2例正常。对核衣壳抗体滴度在第1至7年显著升高的5例SARS患者的胃肠道组织进行抗SARS-CoV N蛋白单克隆抗体免疫组化染色鉴定。这5例患者均经胃镜检查及细胞免疫组化检测。3例血清N蛋白抗体阳性,胃组织黏液腺上皮细胞大部分细胞质免疫组化表达阳性。1例在肠组织浆柱状上皮及间质细胞中也有表达。另外2例血清N蛋白抗体及免疫组化表达均为阴性。5例患者活检结果如下:1例诊断为“胃印戒细胞癌伴直肠多发转移”,1例胃息肉,1例胃窦浅表性胃炎,2例正常。
通过检测14例家庭聚集性病例血清中针对SARS-CoV的特异性IgG、IgM、核衣壳抗体,发现7年时均下降,多数消失。核衣壳抗体滴度与病情有关。证实核衣壳抗体滴度在第7年后显著升高的SARS患者的胃肠道组织中仍存在SARS-CoV。