Eisenbrey A B, Chen J C, Dyer C A, Bernstein J, Poulik M D
Department of Clinical Pathology, William Beaumont Hospital, Royal Oak, Michigan 48073-6769.
J Clin Lab Anal. 1991;5(3):180-6. doi: 10.1002/jcla.1860050306.
A murine monoclonal antibody (E10) was made against cultured cartilage cells. The E10 antibody binding is localized to the surface of cultured cartilage cells in suspension and is present in the cytoplasm in paraffin embedded sections. There is no reactivity with cartilage matrix, or with the matrix of cartilaginous tumors. Reactivity is removed by treatment with trypsin and hyaluronidase, but not by treatment with heparinase, neuraminidase, and chondroitinase. Regeneration of E10 antigen after trypsinization takes 48 hours in chondrocytes in tissue culture. SDS-polyacrylamide gel electrophoresis of an E10 immune precipitate of cultured chondrocytes results in two peaks: one at a very high molecular weight and a small fragment at approximately 250 kd. Specificity has been demonstrated by cytofluorometry, immunofluorescence, and immunohistochemistry, in both frozen and paraffin-embedded tissues. Positive reactivity was seen in cultured cartilage cells, chondrocytes in fetal and adult cartilage, chondrosarcomas, and chordomas. Minimal reactivity was found in a chondromyxoid liposarcoma. Acinar cells of salivary and sweat glands and mast cells in various tissues and tumors were also positive. There was no reactivity with other tissues and tumors, including myxoid and mucinous tumors and epithelial tissues.
制备了一种针对培养软骨细胞的鼠单克隆抗体(E10)。E10抗体结合定位于悬浮培养软骨细胞的表面,在石蜡包埋切片的细胞质中也有存在。它与软骨基质或软骨肿瘤基质无反应性。用胰蛋白酶和透明质酸酶处理可消除反应性,但用肝素酶、神经氨酸酶和软骨素酶处理则不能。在组织培养的软骨细胞中,胰蛋白酶消化后E10抗原的再生需要48小时。培养软骨细胞的E10免疫沉淀物的SDS-聚丙烯酰胺凝胶电泳产生两个峰:一个在非常高的分子量处,另一个在约250kd处的小片段。通过细胞荧光测定法、免疫荧光法和免疫组织化学法在冷冻和石蜡包埋组织中均证实了其特异性。在培养的软骨细胞、胎儿和成人软骨中的软骨细胞、软骨肉瘤和脊索瘤中可见阳性反应性。在软骨黏液样脂肪肉瘤中发现的反应性最低。唾液腺和汗腺的腺泡细胞以及各种组织和肿瘤中的肥大细胞也呈阳性。与其他组织和肿瘤无反应性,包括黏液样和黏液性肿瘤以及上皮组织。