Monoclonal antibodies to murine gamma-interferon which differentially modulate macrophage activation and antiviral activity.
Monoclonal antibodies to murine gamma-interferon which differentially modulate macrophage activation and antiviral activity.
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DOI：
10.4049/jimmunol.134.3.1609
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发表时间：
1985-03
影响因子：
4.4
通讯作者：
R. Schreiber;L. Hicks;A. Celada;N. Buchmeier;P. Gray
中科院分区：
文献类型：
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作者：
R. Schreiber;L. Hicks;A. Celada;N. Buchmeier;P. Gray
通过免疫仓鼠脾细胞与HAT敏感的小鼠骨髓瘤细胞融合，产生了四种抗纯化的重组小鼠γ-干扰素（rIFN-γ）的单克隆IgG抗体。通过使用固定化rIFN-γ的酶联免疫吸附试验（ELISA）或使用可溶性125 I-rIFN-γ和热灭活、固定的金黄色葡萄球菌携带蛋白A的放射免疫测定法确认特异性。竞争结合实验表明，单克隆抗体（MoAb）显示两个不同的表位特异性：一个由H1和H2显示，另一个由H21和H22显示。通过使用鼠-人重组IFN-γ杂交分子，显示H1/H2表位依赖于IFN-γ的氨基末端，而H21/H22表位由羧基末端氨基酸序列形成。单克隆抗体还与天然IFN-γ反应。当与表面结合时，所有四种MoAb，但不是正常仓鼠IgG，去除100%存在于鼠24/G1 T细胞杂交瘤培养物上清液中的抗病毒和MAF活性。在游离溶液中，所有四种抗体都抑制IFN-γ依赖性抗病毒活性，但效率不同。可溶性H21/H22还阻断了所有24/G1衍生的活性，该活性诱导巨噬细胞（MAF）中的非特异性杀肿瘤活性，而H1/H2增强MAF活性。H21或H1的不同抑制或增强活性分别反映了它们抑制或增强125 I-rIFN-γ与巨噬细胞结合的能力。可溶性H21/H22和固相H1/H2抑制100%的MAF，杀微生物，和IA诱导活性的淋巴因子制剂产生的有丝分裂原刺激正常小鼠脾细胞。这些结果有助于建立明确的IFN-γ分子的结构-功能关系，并表明IFN-γ是主要的淋巴因子，负责诱导非特异性杀肿瘤活性和Ia抗原表达，并增强巨噬细胞中的杀微生物活性。
Four monoclonal IgG antibodies to purified, recombinant murine gamma-interferon (rIFN-gamma) have been produced by fusion of immune hamster splenocytes with HAT-sensitive murine myeloma cells. Specificity was confirmed either with an enzyme-linked immunosorbent assay (ELISA) that used immobilized rIFN-gamma or with a radioimmunoassay that employed soluble 125I-rIFN-gamma and heat-killed, fixed Staphylococcus aureus-bearing Protein A. Competition binding experiments suggested that the monoclonal antibodies (MoAb) displayed two distinct epitope specificities: one displayed by H1 and H2, and the other displayed by H21 and H22. By using murine-human recombinant IFN-gamma hybrid molecules, the H1/H2 epitope was shown to depend on the amino-terminus of IFN-gamma, whereas the H21/H22 epitope was formed by the carboxy-terminal amino acid sequence. The MoAb also reacted with natural IFN-gamma. When bound to a surface, all four MoAb, but not normal hamster IgG, removed 100% of the antiviral and MAF activities present in supernatants of cultures of the murine 24/G1 T cell hybridoma. In free solution, all four antibodies inhibited IFN-gamma dependent antiviral activity, but with different efficiencies. Soluble H21/H22 also blocked all of the 24/G1-derived activity that induces nonspecific tumoricidal activity in macrophages (MAF) while H1/H2 enhanced MAF activity. The differential inhibitory or enhancing activities of H21 or H1 reflected their ability to inhibit or enhance binding of 125I-rIFN-gamma to macrophages, respectively. Soluble H21/H22 and solid-phase H1/H2 inhibited 100% of the MAF, microbicidal, and Ia-inducing activities from lymphokine preparations produced by mitogen stimulation of normal murine splenic cells. These results help to establish definitive structure-function relationships for the IFN-gamma molecule, and indicate that IFN-gamma is the primary lymphokine responsible for inducing nonspecific tumoricidal activity and Ia antigen expression, and for enhancing microbicidal activity in macrophages.