Cloning of a tumor-associated antigen: MOv18 and MOv19 antibodies recognize a folate-binding protein.
Cloning of a tumor-associated antigen: MOv18 and MOv19 antibodies recognize a folate-binding protein.
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发表时间：
1991-11
期刊：
影响因子：
11.2
通讯作者：
L. Coney;A. Tomassetti;L. Carayannopoulos;V. Frasca;B. Kamen;M. Colnaghi;V. Zurawski
中科院分区：
文献类型：
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作者：
L. Coney;A. Tomassetti;L. Carayannopoulos;V. Frasca;B. Kamen;M. Colnaghi;V. Zurawski
针对卵巢癌手术标本的膜制剂提出的单抗MOv18和MOv19与大多数测试的非粘液性卵巢恶性肿瘤上存在的表面抗原反应，但不与正常成人组织反应(S.Miotti，S.Canevari，S.Ménard，D.Mezzanzanica，G.Porro，S.M.Pupa，M.Regazzoni，E.Tagaria bue和M.I.Colnaghi，Int.癌症杂志，39：297-303,1987)。经苏云金芽孢杆菌磷脂酶C消化后，从卵巢癌细胞株(IGROV1)的细胞表面释放出一种可溶性糖蛋白，分子量为36-38 kDa。免疫印迹分析表明，纯化的蛋白可与MOv18和MOv19反应，经N-葡聚糖酶处理后得到一种相对分子质量为27 kDa的蛋白。对纯化抗原的氨基末端氨基酸序列进行了测定。该序列与位于人类叶酸结合蛋白NH3末端附近的27个氨基酸的内部延伸高度同源。合成寡核苷酸探针，用于筛选卵巢癌IGROV1、lambda gt11互补DNA文库，获得3个互补DNA克隆。测定了其中一个互补DNA克隆的完整核苷酸序列。该序列与从Caco-2人癌细胞系中获得的叶酸结合蛋白克隆几乎完全相同。此外，还测定了另外两个克隆的5‘非翻译区的核苷酸序列。这三个克隆人的这个区域是不同的。在中国仓鼠卵巢细胞中表达的Caco-2叶酸结合蛋白克隆的产物被MOv18和MOv19抗体识别，证实了该抗原和叶酸结合蛋白是一个整体。此外，与不结合这些抗体的细胞系相比，结合MOv18和MOv19抗体的细胞系表达的叶酸结合蛋白mRNA水平更高。这些结果表明，MOv18和MOv19单抗与至少一种形式的叶酸结合蛋白结合，这种在某些恶性肿瘤中明显过度表达的蛋白可能成为这些抗体免疫治疗的合适靶点。
Monoclonal antibodies MOv18 and MOv19, raised against a membrane preparation of an ovarian carcinoma surgical specimen, react with a surface antigen present on the majority of nonmucinous ovarian malignant tumors tested but not with normal adult tissue (S. Miotti, S. Canevari, S. Ménard, D. Mezzanzanica, G. Porro, S. M. Pupa, M. Regazzoni, E. Tagliabue, and M. I. Colnaghi, Int. J. Cancer, 39: 297-303, 1987). This surface antigen was purified as a soluble glycoprotein (molecular mass, 36-38 kDa) released from the cell surface of an ovarian carcinoma cell line (IGROV1) by digestion with Bacillus thuringiensis phospholipase C. Immunoblotting demonstrated that the purified protein reacted with MOv18 and MOv19 and that treatment of the purified preparation with N-glycanase resulted in a protein with a molecular mass of 27 kDa. The NH3-terminal amino acid sequence of the purified antigen was determined. This sequence is highly homologous to an internal stretch of 27 amino acids located near the NH3 terminus of human folate-binding protein. An oligonucleotide probe was synthesized and used to screen an IGROV1 ovarian carcinoma, lambda gt11 complementary DNA library to obtain three complementary DNA clones. The complete nucleotide sequence of one of these complementary DNA clones was determined. This sequence is nearly identical to that of a folate-binding protein clone obtained from the Caco-2 human carcinoma cell line. In addition, the nucleotide sequence of the 5'-untranslated region of the other two clones was determined. This region of all three clones was different. The product of the Caco-2 folate-binding protein clone expressed in Chinese hamster ovary cells was recognized by the MOv18 and MOv19 antibodies, confirming that the antigen and folate-binding protein are one and the same. Furthermore, a cell line that binds the MOv18 and MOv19 antibodies expressed increased levels of folate-binding protein mRNA compared with a cell line that does not bind these antibodies. These results indicate that the MOv18 and MOv19 monoclonal antibodies bind to at least one form of folate-binding protein and that this protein, which is evidently overexpressed in certain malignant tumors, may provide a suitable target for immunotherapy with these antibodies.