Agostinelli Claudio, Sabattini Elena, Gjørret Jakob Oemar, Righi Simona, Rossi Maura, Mancini Manuela, Piccaluga Pier Paolo, Bacci Francesco, Marafioti Teresa, Bettini Giuliano, Falini Brunangelo, Pileri Stefano A
Departments of Haematology and Clinical Oncology L. and A. Seràgnoli, Pathology and Haematopathology Unit, St. Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 9, Bologna, Italy.
Appl Immunohistochem Mol Morphol. 2010 Dec;18(6):561-72. doi: 10.1097/PAI.0b013e3181e79013.
We describe the newly generated DAK-PAX5 monoclonal antibody raised against a fixation-resistant epitope of the human PAX5/BSAP molecule.
Following Western-blot, absorption, and chess-board titration tests, and optimization of antigen-retrieval and detection methods, DAK-Pax5 was used in parallel with a reference antibody (clone 24) on tissue micro-arrays (TMAs) constructed from normal human and animal tissues and from hematologic and nonhematologic human malignancies. Such TMAs were also tested with an anti-PAX2 antibody.
DAK-Pax5 reacted with normal human and animal B-cells and with 460/473 B-cell non-Hodgkin lymphomas (B-NHLs). All plasmacytomas/plasmablastic tumors (n=13) and T/NK-cell neoplasms (n=264) turned out consistently negative as did acute myelogenous leukaemias (n=19) except 2 carrying t(8;21). Positivity was found in 6/6 and 155/169 lymphocyte predominant and classical HLs, respectively, although the staining intensity varied through cases. Among 521 nonhematologic malignancies, DAK-Pax5 reacted with 22/399 carcinomas (4/11 neuroendocrine, 2/4 Merkel-cell, 4/21 prostatic, 1/11 urothelial, 1/26 renal, 2/12 cervical squamous-cell, 3/13 ovarian, and 5/75 colonic). When compared with clone 24, DAK-Pax5 produced a stronger positivity in most if not all B-NHLs and HLs. No cross-reactivity with the anti-PAX2 antibody was recorded.
DAK-Pax5 represents a new reliable tool for diagnostics and research.
我们描述了新产生的DAK-PAX5单克隆抗体，该抗体针对人PAX5/BSAP分子的一个抗固定表位产生。
经过蛋白质免疫印迹、吸收和棋盘滴定试验，并优化抗原修复和检测方法后，将DAK-Pax5与一种参考抗体（克隆24）同时应用于由正常人和动物组织以及血液系统和非血液系统人类恶性肿瘤构建的组织微阵列（TMA）。这些TMA也用抗PAX2抗体进行了检测。
DAK-Pax5与正常人和动物的B细胞以及460/473例B细胞非霍奇金淋巴瘤（B-NHL）发生反应。所有浆细胞瘤/浆母细胞瘤（n=13）和T/NK细胞肿瘤（n=264）均始终呈阴性，急性髓性白血病（n=19）除2例携带t(8;21)外也呈阴性。在6/6例淋巴细胞为主型和155/169例经典型霍奇金淋巴瘤（HL）中均发现阳性，尽管各病例的染色强度有所不同。在521例非血液系统恶性肿瘤中，DAK-Pax5与22/399例癌发生反应（4/11例神经内分泌癌、2/4例默克尔细胞癌、4/21例前列腺癌、1/11例尿路上皮癌）。1/26例肾癌、2/12例宫颈鳞状细胞癌、3/13例卵巢癌和5/75例结肠癌）。与克隆24相比，DAK-Pax5在大多数（如果不是全部）B-NHL和HL中产生更强的阳性反应。未记录到与抗PAX2抗体的交叉反应。
DAK-Pax5是诊断和研究的一种新的可靠工具。
