MONOCLONAL ANTI-IGM REGULATION OF HUMAN B CELL FUNCTION

单克隆抗 IGM 对人 B 细胞功能的调节

• 批准号: 3287455
• 负责人: Patricia K. Mongini
• 金额: $ 13.1万
• 依托单位: HOSPITAL FOR JOINT DISEASES ORTHO INST
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3287455
• 关键词: B lymphocyte; blocking antibody; cell differentiation; cell membrane; crosslink; histocompatibility antigens; human tissue; hybridomas; immunoglobulin M; immunoglobulins; ion exchange chromatography; leukocyte activation /transformation; monoclonal antibody; radioimmunoassay; radiotracer; rheumatoid factor; stoichiometry

The proposed study will investigate the nature of the ligand interaction with membrane IgM which is needed for B lymphocyte stimulation. This will be accomplished through the use of a panel of anti-human IgM monoclonal antibodies (MoAbs) which differ in affinity and/or epitope specificity. Using these defined anti-Ig ligands, an explanation for the diverse regulatory effects of anti-Ig ligands which have been observed in the past will be sought. These studies may elucidate the factors that determine whether or not autologous anti-Ig constant region antibodies (rheumatoid factors) can regulate the function of B cells to which they bind. The different mouse anti-human IgM MoAbs will be compared in their ability to a) induce resting B cells to proliferate, b) affect the proliferation and differentiation of B cells stimulated with other mitogens, and c) stimulate early changes in B cells which precede the S phase of the cell cycle. Once sets of stimulatory and non-stimulatory (perhaps inhibitory) MoAbs have been identified, explanations for the functional differences in these ligands will be sought. This will involve comparing stimulatory and non-stimulatory MoAbs and their F(ab')2 fragments for a) affinity and stoichiometry of binding to IgM and b) epitope specificity. The epitope specificity of the MoAbs will be investigated by competitive inhibition studies and by analysis of the ability of each MoAb to bind to chemically modified IgM, IgM fragments, and synthesized peptides which have been predicted by hydrophilicity and conformational analysis as likely to constitute immunogenic epitopes on the human IgM molecule. In addition, the nature of membrane molecule crosslinkage which is necessary to stimulate B cell proliferation will be studied. This will involve comparing the mitogenic capacity of a divalent anti-IgM MoAb to a hybrid antibody which consists of one F(ab') arm directed to IgM and one F(ab') arm directed to a human major histocompatibility (MHC) antigen.

拟议的研究将研究配体相互作用的性质 与B淋巴细胞刺激所需的膜IgM。 这会 可以通过使用一组抗人IgM单克隆来实现 抗体（MOAB）的亲和力和/或表位特异性不同。 使用这些定义的抗gig配体，这是对多样的解释 过去观察到的抗gig配体的调节作用 将被寻求。 这些研究可以阐明确定的因素 自体抗gig常数区域抗体（类风湿）是否 因素）可以调节其结合的B细胞的功能。 将比较不同的小鼠抗人IGM摩押仪的能力 a）诱导静息B细胞增殖，b）影响增殖 以及用其他有差点刺激的B细胞的分化，C） 刺激在细胞S相之前的B细胞的早期变化 循环。 曾经有一组刺激性和非刺激性（也许是抑制性） 已经确定了摩押，对功能差异的解释 这些配体将被寻求。 这将涉及比较刺激和 非刺激的摩押及其f（ab'）2片段a）亲和力和 与IgM和B）表位特异性结合的化学计量法。 表位 将通过竞争性抑制来研究摩押的特异性 研究和通过分析每个摩押仪与化学结合的能力 修改的IgM，IgM片段和合成的肽已 通过亲水性和构象分析预测 构成人IgM分子的免疫原性表位。 另外，膜分子交联的性质是 将研究刺激B细胞增殖所必需的。 这会 涉及比较二价抗IGM摩押的有丝分裂能力 由一个针对IGM的F（AB'）手臂组成的混合抗体和一种 F（AB'）手臂针对人类主要的组织相容性（MHC）抗原。