Isolation and characterization of monoclonal antibodies directed to rat interferon-gamma.

Lymphokine research Pub Date : 1989-01-01
P H van der Meide, A H Borman, H G Beljaars, M A Dubbeld, C A Botman, H Schellekens
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Abstract

Eight stable hybridoma cell lines producing monoclonal antibodies reactive with rat interferon-gamma have been generated. The antibodies produced belong to three different immunoglobulin isotypes: IgG1 (4 monoclonal antibodies, designated DB-1, DB-10, DB-12 and DB-13), IgG2a (3 monoclonal antibodies, DB-9, DB-14 and DB-16) and IgA (1 monoclonal antibody, DB-2). The antibodies were characterized in terms of epitope specificity and reactivity with rat, mouse and human interferon-gamma. Three antibodies (DB-1, -2 and -14) show high antiviral neutralizing activity against natural and recombinant DNA derived rat interferon-gamma, whereas the others have low (DB-10, -12, -13 and -16) or no (DB-9) detectable activity. Two antibodies (DB-1 and -2) effectively bind and neutralize mouse interferon-gamma, one antibody (DB-14) exhibits some cross-reactivity and the others show no reactivity towards the mouse lymphokine. None of the antibodies reacts with human interferon-gamma. All antibodies recognize immunoblotted recombinant rat interferon-gamma, although substantial variation in the immunoreactivity existed. Competition binding experiments reveal that the antibodies are directed to three spatially distinct sites on the rat lymphokine. Two non-competing monoclonal antibodies were selected and used for the development of a specific enzyme-linked immunosorbent assay for the detection of rat interferon-gamma in biological fluids.

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大鼠γ干扰素单克隆抗体的分离与鉴定。
制备了8株稳定的杂交瘤细胞系,产生与大鼠γ干扰素反应的单克隆抗体。产生的抗体属于三种不同的免疫球蛋白同型:IgG1(4种单克隆抗体,指定为DB-1, DB-10, DB-12和DB-13), IgG2a(3种单克隆抗体,指定为DB-9, DB-14和DB-16)和IgA(1种单克隆抗体,DB-2)。根据表位特异性和对大鼠、小鼠和人γ干扰素的反应性对抗体进行了表征。三种抗体(DB-1、-2和-14)对天然和重组DNA来源的大鼠干扰素γ具有高抗病毒中和活性,而其他抗体的活性较低(DB-10、-12、-13和-16)或无活性(DB-9)。两种抗体(DB-1和-2)有效结合并中和小鼠干扰素- γ,一种抗体(DB-14)表现出一定的交叉反应性,而其他抗体对小鼠淋巴因子无反应性。这些抗体都不会与人γ干扰素发生反应。所有抗体都能识别免疫印迹重组大鼠干扰素- γ,尽管免疫反应性存在实质性差异。竞争结合实验表明,抗体被定向到大鼠淋巴因子上的三个空间不同的位点。选择两种非竞争单克隆抗体,用于开发一种特异性酶联免疫吸附试验,用于检测生物体液中的大鼠干扰素。
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