Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.
Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.
复制标题
两种新的单克隆抗体 (LN-1、LN-2) 在 B5 福尔马林固定、石蜡包埋的组织中具有反应性，其中包括滤泡中心和外套区人 B 淋巴细胞和衍生肿瘤。
作者：
A. Epstein;R. Marder;J. Winter;R. Fox
分别利用来自商陆丝裂原刺激的外周血淋巴细胞和弥漫性组织细胞淋巴瘤 SU-DHL-4 细胞的细胞提取物，制备了两种与 B5 福尔马林固定、石蜡包埋的组织切片中的 B 淋巴细胞发生反应的单克隆抗体 (LN-1、LN-2)。两种单克隆抗体最初都是通过间接免疫荧光筛选技术对多聚甲醛-丙酮固定的细胞制剂进行鉴定的。对 36 种充分表征的人类淋巴瘤和白血病细胞系进行的特异性筛选表明，LN-1 和 LN-2 均可对 B 细胞谱系的细胞系进行染色，但与 T 细胞或骨髓来源的细胞系不发生反应（其中一种例外）。发现空细胞急性淋巴细胞白血病细胞系为LN-2+但LN-1-。通过使用间接免疫荧光技术，这些试剂的 B 细胞特异性在 15 例淋巴瘤和 17 例白血病活检标本上得到证实。对 B5 固定、石蜡包埋的人淋巴组织切片进行免疫过氧化物酶染色显示，LN-1 与生发中心细胞的细胞膜和细胞质结合，而 LN-2 对生发中心和套区 B 淋巴细胞以及滤泡间组织细胞和胸腺髓质树突细胞的核膜和细胞质进行染色。两种单克隆抗体均未能对皮质胸腺细胞、淋巴结 T 细胞、外周血 T 细胞和骨髓细胞进行染色。对 20 个非淋巴人体器官和组织的免疫过氧化物酶染色显示，LN-1 与骨髓的红细胞前体、支气管的纤毛上皮细胞、肾脏的远端管状细胞以及来自包括乳房和前列腺在内的多个器官的导管细胞发生阳性反应。相比之下，LN-2 与所有人类非淋巴器官和组织（包括骨髓）均不发生反应。一组 26 个实体瘤细胞系的间接免疫荧光染色显示，LN-1 与大多数上皮来源的细胞系、胶质母细胞瘤和星形细胞瘤有反应，但与神经母细胞瘤、小细胞肺癌和肉瘤没有反应。通过这些技术，LN-2 与 26 个实体瘤细胞系中的 25 个不发生反应。免疫生化研究表明，LN-1 识别细胞表面唾液酸抗原，而 LN-2 则针对 35,000 道尔顿的核膜蛋白。由于 LN-1 和 LN-2 对 B 细胞肿瘤具有高度特异性，并且能够对 B5 固定、石蜡包埋的组织进行染色，因此它们是诊断和分类人类淋巴瘤和白血病的有用试剂。
Two monoclonal antibodies (LN-1, LN-2) reactive with B lymphocytes in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by utilizing cell extracts from pokeweed mitogen-stimulated peripheral blood lymphocytes and diffuse histiocytic lymphoma SU-DHL-4 cells, respectively. Both monoclonal antibodies were initially identified by indirect immunofluorescence screening techniques on paraformaldehyde-acetone-fixed cell preparations. Specificity screens with 36 well-characterized human lymphoma and leukemia cell lines showed that both LN-1 and LN-2 stained cell lines of B cell lineage but were unreactive with those of T cell or, with one exception, myeloid derivation. Null cell acute lymphoblastic leukemia cell lines were found to be LN-2+ but LN-1-. The B cell specificity of these reagents was confirmed on 15 lymphoma and 17 leukemia biopsy specimens by using indirect immunofluorescence techniques. Immunoperoxidase staining of sections from B5-fixed, paraffin-embedded human lymphoid tissues showed that LN-1 bound to the cell membrane and cytoplasm of germinal center cells whereas LN-2 stained the nuclear membrane and cytoplasm of germinal center and mantle zone B lymphocytes as well as interfollicular histiocytes and thymic medullary dendritic cells. Both monoclonal antibodies failed to stain cortical thymocytes, lymph node T cells, and peripheral blood T and myeloid cells. Immunoperoxidase staining of 20 nonlymphoid human organs and tissues revealed that LN-1 reacted positively with red blood cell precursors of the bone marrow, ciliated epithelial cells of the bronchus, distal tubular cells of the kidney, and ductal cells from several organs including the breast and prostate. In contrast, LN-2 was unreactive with all human nonlymphoid organs and tissues including the bone marrow. Indirect immunofluorescence staining of a panel of 26 solid tumor cells lines showed that LN-1 was reactive with the majority of epithelium-derived cell lines, glioblastomas, and astrocytomas but was unreactive with neuroblastomas, small cell carcinoma of the lung, and sarcomas. LN-2 was unreactive with 25 of 26 of the solid tumor cell lines by these techniques. Immunobiochemical studies have shown that LN-1 recognizes a cell surface sialoantigen whereas LN-2 is directed against a 35,000 dalton nuclear membrane protein. Because of their high specificity for B cell tumors and their ability to stain B5-fixed, paraffin-embedded tissues, LN-1 and LN-2 are useful reagents for the diagnosis and classification of the human lymphomas and leukemias.