A panel of monoclonal antibodies recognizing GPI-anchored ADP-ribosyltransferase ART4, the carrier of the Dombrock blood group antigens
A panel of monoclonal antibodies recognizing GPI-anchored ADP-ribosyltransferase ART4, the carrier of the Dombrock blood group antigens
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DOI：
10.1016/j.cellimm.2005.08.008
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发表时间：
2005-07-01
影响因子：
4.3
通讯作者：
Koch-Nolte, F
中科院分区：
文献类型：
--
作者：
Parusel, I;Kahl, S;Koch-Nolte, F
ART 4（CD 297）是毒素相关ADP-核糖基转移酶（ART）家族的成员，并且是Dombrock血型同种异体抗原（Do）的携带者。分别用人Do/ART 4转染细胞免疫小鼠和用人Do/ART 4 cDNA免疫大鼠，获得两种小鼠单克隆抗体（米马-52和米马-53）和两种大鼠单克隆抗体（NONI-B4和NONI-1363）。通过FACS分析，所有四种mAb识别Do/ART 4转染的Jurkat细胞，但不识别未转染的细胞。用PI-PLC处理细胞后，这些mAb对Do/ART 4转染细胞的染色减少，证实Do/ART 4通过与糖基磷脂酰肌醇连接而锚定在细胞膜中，如从其氨基酸序列预测的那样。这四种mAb不与戈伊（a-）（东布罗克无效）红细胞发生反应，但凝集其他红细胞。流式细胞仪分析表明，所有mAb均与红细胞反应，与外周血单核细胞和脾巨噬细胞反应较弱。而不是B淋巴细胞或T淋巴细胞。该单克隆抗体与人脐静脉内皮细胞和嗜碱性白血病KU-812也有微弱的反应。免疫组化显示扁桃体切片上上皮细胞和内皮细胞染色。在FACS分析中，NONI-1314与米马-52竞争结合Do/ART 4转染的细胞和红细胞，而NONI-1363与米马-53竞争。两种mAb均不与小鼠ART 4转染细胞反应，但NONI-B63和米马-53与小鼠/人ART 4嵌合体反应，表明这些mAb识别的表位位于蛋白质的C-末端一半。(c)2005年爱思唯尔公司All rights reserved.
ART4 (CD297) is a member of the family of toxin-related ADP-ribosyltransferases (ARTs) and is the carrier of the Dombrock blood group alloantigens (Do). Two mouse monoclonal antibodies (MIMA-52 and MIMA-53), and two rat monoclonal antibodies (N0NI-B4 and NONI-1363) were obtained following immunization of mice with human Do/ART4-transfected cells and of rats with human Do/ART4 cDNA, respectively. All four mAbs recognize Do/ART4-transfected Jurkat cells but not untransfected cells by FACS analysis. Staining of Do/ART4-transfected cells by these mAbs was reduced following treatment of cells with PI-PLC, confirming that Do/ART4 is anchored in the cell membrane by linkage to glycosylphosphatidylinositol as predicted from its amino acid sequence. The four mAbs did not react with Gy(a-) (Dombrock null) erythrocytes but agglutinated other red blood cells. By flow cytometric analysis, all mAbs reacted prominently with erythrocytes, and weakly with peripheral blood monocytes and splenic macrophages. but not with B-lymphocytes or T-lymphocytes. The mAbs reacted weakly also with human umbilical vein endothelial cells and the basophilic leukemia KU-812. Immunohistology revealed staining of epithelia and endothelia, on sections of tonsils. In FACS analyses NONI-1314 competed with MIMA-52 for binding to Do/ART4-transfected cells and erythrocytes, whereas NONI-1363 competed with MIMA-53. Neither of the mAbs reacted with mouse ART4-transfected cells, but NONI-B63 and MIMA-53 did react with a mouse/human ART4 chimera, indicating that the epitope recognized by these mAbs lies in the C-terminal half of the protein. (c) 2005 Elsevier Inc. All rights reserved.