A novel monoclonal antibody for detection of galectin-9 in tissue sections: application to human tissues infected by oncogenic viruses.
A novel monoclonal antibody for detection of galectin-9 in tissue sections: application to human tissues infected by oncogenic viruses.
复制标题
DOI：
10.1186/1750-9378-7-16
复制
发表时间：
2012-07-17
影响因子：
3.7
通讯作者：
Busson P
中科院分区：
文献类型：
--
作者：
Barjon C;Niki T;Vérillaud B;Opolon P;Bedossa P;Hirashima M;Blanchin S;Wassef M;Rosen HR;Jimenez AS;Wei M;Busson P
Galectin-9是一种具有免疫抑制作用的哺乳动物凝集素。据报道，在两种与人类病毒相关的疾病中，Galectin-9的过度产生与Epstein-Barr病毒相关的慢性丙型肝炎和鼻咽癌有关。本研究的目的是制备新的针对Galectin-9的单抗，以提高Galectin-9在临床标本中的检测水平，特别是在免疫组织化学分析的组织切片上。用重组Galectin-9 C末端部分(长链191~355残基)免疫小鼠，脾细胞与Sp2/0细胞半固体融合，获得杂交瘤细胞。用酶联免疫吸附试验、表位定位、免疫印迹和免疫组织化学等方法对单抗进行鉴定。我们选择了7个杂交瘤细胞产生与我们的重组c-末端Galectin-9反应的抗体。其中5个与表位“TPAIPPMMYPHPA”(所有异构体共有的，长异构体的210-222个残基)反应，并通过蛋白质印迹分析染色了Galectin-9的所有三个异构体。其中1G3在免疫组织化学中表现出很好的敏感性和特异性。通过使用1G3，我们可以证实鼻咽癌Epstein-Barr病毒阳性的恶性细胞强烈且持续表达Galectin-9。在大多数标本中，胞浆和胞核均可见特异性染色。在人类丙型或乙型肝炎病毒感染患者的肝活检组织中也检测到Galectin-9，该蛋白不仅在炎性白细胞和Kupffer细胞中表达，而且在肝细胞中也有表达。相比之下，在未感染的肝脏样本中几乎不存在Galectin-9。1G3单抗将成为评估Galectin-9表达和分布的有力工具，特别是在与致癌病毒相关的疾病中。
Galectin-9 is a mammalian lectin which possesses immunosuppressive properties. Excessive production of galectin-9 has been reported in two types of human virus-associated diseases chronic hepatitis C and nasopharyngeal carcinoma associated to the Epstein-Barr virus. The objective of this study was to produce new monoclonal antibodies targeting galectin-9 in order to improve its detection in clinical samples, especially on tissue sections analysed by immunohistochemistry. Hybridomas were produced through immunization of mice with the recombinant c-terminus part of galectin-9 (residues 191 to 355 of the long isoform) and semi-solid fusion of spleen cells with Sp2/0 cells. Monoclonal antibodies were characterized using ELISA, epitope mapping, western blot and immunohistochemistry. We selected seven hybridomas producing antibodies reacting with our recombinant c-terminus galectin-9 in ELISA. Five of them reacted with the epitope “TPAIPPMMYPHPA” (common to all isoforms, residues 210 to 222 of the long isoform) and stained all three isoforms of galectin-9 analysed by western blot. One of them, 1G3,demonstrated very good sensitivity and specificity when used for immunohistochemistry. Using 1G3, we could confirm the intense and constant expression of galectin-9 by Epstein-Barr virus positive malignant cells from nasopharyngeal carcinomas. In most samples, specific staining was detected in both cytoplasm and nuclei. Galectin-9 was also detected in liver biopsies from patients infected by the human hepatitis C or B viruses with expression not only in inflammatory leucocytes and Kupffer cells, but also in hepatocytes. In contrast, galectin-9 was virtually absent in non-infected liver specimens. The 1G3 monoclonal antibody will be a powerful tool to assess galectin-9 expression and distribution especially in diseases related to oncogenic viruses.
登录
查看更多内容
DOI：
10.1073/pnas.1107087108
发表时间：
2011-11-22
影响因子：
11.1
作者：
Sharma, Shalini;Sundararajan, Aarthi;Rouse, Barry T.
通讯作者：
Rouse, Barry T.
DOI：
10.1084/jem.20060210
发表时间：
2006-06-12
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Koguchi K;Anderson DE;Yang L;O'Connor KC;Kuchroo VK;Hafler DA
通讯作者：
Hafler DA
影响因子：
4.8
作者：
Niki, Toshiro;Tsutsui, Shoko;Hirashima, Mitsuomi
通讯作者：
Hirashima, Mitsuomi
影响因子：
5
作者：
Stancic, M.;van Horssen, J.;Baron, W.
通讯作者：
Baron, W.
影响因子：
5.1
作者：
Lee, Jaejoon;Park, Eun-Jung;Cha, Hoon-Suk
通讯作者：
Cha, Hoon-Suk