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与人类嗜T淋巴细胞病毒I型(HTLV-I)的三种核心蛋白(p24、p19和p15)相关的抗原及其细胞内定位,由单克隆抗体确定。

Antigens related to three core proteins of HTLV-I (p24, p19 and p15) and their intracellular localizations, as defined by monoclonal antibodies.

作者信息

Tanaka Y, Lee B, Inoi T, Tozawa H, Yamamoto N, Hinuma Y

出版信息

Int J Cancer. 1986 Jan 15;37(1):35-42. doi: 10.1002/ijc.2910370107.

Abstract

Three distinct monoclonal antibodies (MAbs) specific for human T-cell leukemia virus type-I (HTLV-I) core proteins with molecular weights of 24 kDa (p24), p19 or p15 were produced, characterized and compared. These antibodies were named NOR-1 (anti-p24, IgG2a), GIN-7 (anti-p19, IgG2b) and FR-45 (anti-p15, IgG2a). Immunofluorescence assay showed that they reacted specifically with methanol-fixed cells of virus-bearing cell lines, and that only GIN-7 bound, albeit weakly, to the surface of a small percentage of viable cells. Like natural antibodies to HTLV-I in human serum, GIN-7 stained the fixed cells brightly and diffusely, and gave more intense fluorescence than NOR-1 and FR-45, which stained restricted areas of the cells. NOR-1, GIN-7 and FR-45 specifically precipitated core proteins p24, p19 and p15, respectively, from a lysate of HTLV-IMT-2 labelled with 35S-cysteine. NOR-1 precipitated p53, p36, and p24, GIN-7 precipitated p53, p32, p28 and p19, and FR-45 precipitated p53, p36, and p15 from a lysate of 35S-cysteine-labelled MT-2 cells. GIN-7 also precipitated p32, p28 and p19 from a lysate of MT-2 cells, labelled by surface iodination, but NOR-1 and FR-45 did not detect any proteins in this lysate. GIN-7 also detected p28 in 3H-glucosamine-labelled MT-2 cells. Antibody binding competition assay showed that the sera of ATL patients significantly interfered with the binding of NOR-1 and GIN-7 but not with that of FR-45, to antigens of disrupted virus of MT-2 cells. This complete set of MAbs against the HTLV-I gag gene products is useful for biological and functional studies of the HTLV-I core proteins.

摘要

制备、鉴定并比较了三种分别针对分子量为24 kDa(p24)、p19或p15的人T细胞白血病病毒I型(HTLV-I)核心蛋白的独特单克隆抗体(MAb)。这些抗体分别命名为NOR-1(抗p24,IgG2a)、GIN-7(抗p19,IgG2b)和FR-45(抗p15,IgG2a)。免疫荧光分析表明,它们与携带病毒的细胞系的甲醇固定细胞发生特异性反应,并且只有GIN-7与一小部分活细胞的表面结合,尽管结合较弱。与人类血清中针对HTLV-I的天然抗体一样,GIN-7能明亮且弥散地对固定细胞进行染色,并且比NOR-1和FR-45产生更强的荧光,后两者仅对细胞的特定区域进行染色。NOR-1、GIN-7和FR-45分别从用35S-半胱氨酸标记的HTLV-IMT-2裂解物中特异性沉淀出核心蛋白p24、p19和p15。NOR-1从35S-半胱氨酸标记的MT-2细胞裂解物中沉淀出p53、p36和p24,GIN-7沉淀出p53、p32、p28和p19,FR-45沉淀出p53、p36和p15。GIN-7还从经表面碘化标记的MT-2细胞裂解物中沉淀出p32、p28和p19,但NOR-1和FR-45在该裂解物中未检测到任何蛋白质。GIN-7还在3H-葡萄糖胺标记的MT-2细胞中检测到p28。抗体结合竞争试验表明,成人T细胞白血病(ATL)患者的血清显著干扰NOR-1和GIN-7与MT-2细胞破碎病毒抗原的结合,但不干扰FR-45的结合。这一套完整的针对HTLV-I gag基因产物的单克隆抗体对于HTLV-I核心蛋白的生物学和功能研究很有用。

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