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与人类B淋巴细胞发生反应的单克隆抗体的产生与特性鉴定

Generation and characterization of monoclonal antibodies reactive with human B lymphocytes.

作者信息

Mittler R S, Talle M A, Carpenter K, Rao P E, Goldstein G

出版信息

J Immunol. 1983 Oct;131(4):1754-61.

PMID:6413577
Abstract

Four novel monoclonal antibodies directed at determinants on human B lymphocytes were produced and characterized by flow cytometry using indirect immunofluorescence analysis. The antibodies, OKB1, OKB2, OKB4, and OKB7, define antigens not previously described on human B lymphocytes using B cell reactive monoclonal antibodies or standard B cell phenotypic markers. OKB4 [immunoglobulin M (IgM)] and OKB7 (IgG2a) were reactive with greater than 95% of all surface membrane immunoglobulin positive (Smlg+) lymphocytes and were restricted to the B cell lineage. OKB2 (IgG1) likewise reacted with greater than 95% of all Smlg+ lymphocytes; however, the antigen was also expressed on all granulocytes. In contrast to OKB2, OKB4, and OKB7, OKB1 reacted with a variable percentage of normal peripheral B cells (approximately 70 to 95%). Functional studies employing polyclonally activated B cells in a pokeweed mitogen (PWM) driven system and the reverse hemolytic plaque assay for the quantitation of Ig secreting plasma cells demonstrated that OKB1 and OKB2 inhibited the generation of plaque-forming cells (PFC) when added to cultures at day 0. OKB4 had a marginal inhibitory effect, whereas OKB7 consistently enhanced the PFC response. Immunoprecipitation studies with OKB1, OKB4, and OKB7 demonstrated that these antibodies precipitated antigens of approximately 168,000, 87,000, and 175,000 m.w., respectively. The OKB2 antigen is presently under study.

摘要

制备了四种针对人B淋巴细胞决定簇的新型单克隆抗体,并通过间接免疫荧光分析的流式细胞术进行了表征。使用B细胞反应性单克隆抗体或标准B细胞表型标志物,抗体OKB1、OKB2、OKB4和OKB7定义了人B淋巴细胞上以前未描述的抗原。OKB4(免疫球蛋白M(IgM))和OKB7(IgG2a)与所有表面膜免疫球蛋白阳性(Smlg+)淋巴细胞的95%以上发生反应,并且仅限于B细胞谱系。OKB2(IgG1)同样与所有Smlg+淋巴细胞的95%以上发生反应;然而,该抗原也在所有粒细胞上表达。与OKB2、OKB4和OKB7不同,OKB1与可变比例的正常外周B细胞发生反应(约70%至95%)。在商陆有丝分裂原(PWM)驱动的系统中使用多克隆激活的B细胞进行的功能研究以及用于定量Ig分泌浆细胞的反向溶血空斑试验表明,在第0天添加到培养物中时,OKB1和OKB2抑制空斑形成细胞(PFC)的产生。OKB4具有轻微的抑制作用,而OKB7始终增强PFC反应。用OKB1、OKB4和OKB7进行的免疫沉淀研究表明,这些抗体分别沉淀了分子量约为168,000、87,000和175,000的抗原。OKB2抗原目前正在研究中。

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Generation and characterization of monoclonal antibodies reactive with human B lymphocytes.与人类B淋巴细胞发生反应的单克隆抗体的产生与特性鉴定
J Immunol. 1983 Oct;131(4):1754-61.
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