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Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein.
Veterinary Immunology and Immunopathology ( IF 1.4 ) Pub Date : 2019-09-09 , DOI: 10.1016/j.vetimm.2019.109939
Ann-Kathrin Haverkamp 1 , Berend J Bosch 2 , Ingo Spitzbarth 3 , Annika Lehmbecker 3 , Nigeer Te 4 , Albert Bensaid 4 , Joaquim Segalés 5 , Wolfgang Baumgärtner 3
Affiliation  

Middle East respiratory syndrome (MERS) represents an important respiratory disease accompanied by lethal outcome in one third of human patients. In recent years, several investigators developed protective antibodies which could be used as prophylaxis in prospective human epidemics. In the current study, eight human monoclonal antibodies (mAbs) with neutralizing and non-neutralizing capabilities, directed against different epitopes of the MERS-coronavirus (MERS-CoV) spike (MERS-S) protein, were investigated with regard to their ability to immunohistochemically detect respective epitopes on formalin-fixed paraffin-embedded (FFPE) nasal tissue sections of MERS-CoV experimentally infected alpacas. The most intense immunoreaction was detected using a neutralizing antibody directed against the receptor binding domain S1B of the MERS-S protein, which produced an immunosignal in the cytoplasm of ciliated respiratory epithelium and along the apical membranous region. A similar staining was obtained by two other mAbs which recognize the sialic acid-binding domain and the ectodomain of the membrane fusion subunit S2, respectively. Five mAbs lacked immunoreactivity for MERS-CoV antigen on FFPE tissue, even though they belong, at least in part, to the same epitope group. In summary, three tested human mAbs demonstrated capacity for detection of MERS-CoV antigen on FFPE samples and may be implemented in double or triple immunohistochemical methods.

中文翻译:

使用针对穗蛋白不同表位的人单克隆抗体通过免疫组织化学检测福尔马林固定石蜡包埋的羊驼鼻组织上的MERS-CoV抗原。

中东呼吸综合征(MERS)是一种重要的呼吸系统疾病,其三分之一的人类患者伴有致命的后果。近年来,几位研究人员开发了可用于预防人类流行病的保护性抗体。在当前的研究中,针对MERS冠状病毒(MERS-CoV)刺突(MERS-S)蛋白的不同表位,研究了八种具有中和和非中和能力的人单克隆抗体(mAb),免疫组化检测MERS-CoV实验感染的羊驼的福尔马林固定石蜡包埋(FFPE)鼻腔组织切片上的各个表位。使用针对MERS-S蛋白的受体结合结构域S1B的中和抗体检测到最强烈的免疫反应,在纤毛的呼吸道上皮的细胞质中以及沿顶端的膜状区域产生免疫信号。通过另外两个分别识别膜融合亚基S2的唾液酸结合结构域和胞外结构域的mAb,获得了相似的染色。五个mAb对FFPE组织上的MERS-CoV抗原缺乏免疫反应性,即使它们至少部分属于同一表位组也是如此。总之,三种测试的人类单克隆抗体证明了具有检测FFPE样品上的MERS-CoV抗原的能力,并且可以采用双重或三重免疫组化方法实施。通过另外两个分别识别膜融合亚基S2的唾液酸结合结构域和胞外结构域的mAb,获得了相似的染色。五个mAb对FFPE组织上的MERS-CoV抗原缺乏免疫反应性,即使它们至少部分属于同一表位组也是如此。总之,三种测试的人类单克隆抗体证明了具有检测FFPE样品上的MERS-CoV抗原的能力,并且可以采用双重或三重免疫组化方法实施。通过另外两个分别识别膜融合亚基S2的唾液酸结合结构域和胞外结构域的mAb,获得了相似的染色。五个mAb对FFPE组织上的MERS-CoV抗原缺乏免疫反应性,即使它们至少部分属于同一表位组也是如此。总之,三种测试的人类单克隆抗体证明了具有检测FFPE样品上的MERS-CoV抗原的能力,并且可以采用双重或三重免疫组化方法实施。
更新日期:2019-11-01
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