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通过重组DNA产生的人嗜T淋巴细胞病毒III型肽与艾滋病患者的血清具有免疫反应性。

An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS.

作者信息

Chang N T, Huang J, Ghrayeb J, McKinney S, Chanda P K, Chang T W, Putney S, Sarngadharan M G, Wong-Staal F, Gallo R C

出版信息

Nature. 1985;315(6015):151-4. doi: 10.1038/315151a0.

Abstract

Human T-cell lymphotropic retrovirus type III (HTLV-III), also called lymphadenopathy-associated virus (LAV), has been identified as the aetiological agent of acquired immune deficiency syndrome (AIDS). The sera of most patients with AIDS or AIDS-related complexes, and of asymptomatic individuals infected with HTLV-III, contain antibodies against antigens of HTLV-III. The characterization of these antibodies and their corresponding viral antigens is important not only for understanding immunity against HTLV-III and the pathology of AIDS, but also for the development of diagnostic methods and preventive vaccine for AIDS. Following the successful establishment of a long-term T-cell line permissive for HTLV-III replication, large quantities of virus have been produced, facilitating the purification of viral proteins and the development of mouse monoclonal antibodies against several viral antigens. More recently, the structure of HTLV-III proviral DNA has been elucidated. We now report the production, by genetic engineering methods, of a peptide encoded by a gene segment of HTLV-III. A 1.1-kilobase (kb) EcoRI DNA segment from an isolate of HTLV-III was inserted into a lpp and lac promoter-coupled expression vector, pIN-III-ompA. Escherichia coli transformants of this plasmid produced a peptide of relative molecular mass (Mr) 15,000 (15K) which was strongly immunoreactive with anti-HTLV-III antibodies present in sera from AIDS patients. Lysates of the clones expressing this 15K peptide inhibited the reactivity of the p31 virion protein with AIDS sera, suggesting that it is a fragment of the viral p31 protein. The peptide reacted with sera from all 20 AIDS patients but none of the 8 normal controls tested. These results suggest that the peptide may be useful for detecting anti-HTLV-III antibodies in blood samples.

摘要

人类嗜T细胞病毒III型(HTLV - III),也称为淋巴结病相关病毒(LAV),已被确认为获得性免疫缺陷综合征(AIDS)的病原体。大多数艾滋病患者或艾滋病相关综合征患者以及感染HTLV - III的无症状个体的血清中,都含有针对HTLV - III抗原的抗体。这些抗体及其相应病毒抗原的特性不仅对于理解针对HTLV - III的免疫和艾滋病的病理十分重要,而且对于艾滋病诊断方法和预防性疫苗的开发也很重要。在成功建立了允许HTLV - III复制的长期T细胞系之后,已生产出大量病毒,这有助于病毒蛋白的纯化以及针对几种病毒抗原的小鼠单克隆抗体的开发。最近,HTLV - III前病毒DNA的结构已被阐明。我们现在报告通过基因工程方法生产的一种由HTLV - III基因片段编码的肽。将来自HTLV - III分离株的1.1千碱基(kb)的EcoRI DNA片段插入到一个与lpp和lac启动子偶联的表达载体pIN - III - ompA中。该质粒的大肠杆菌转化体产生了一种相对分子质量(Mr)为15,000(15K)的肽,它与艾滋病患者血清中存在的抗HTLV - III抗体具有强烈的免疫反应性。表达这种15K肽的克隆裂解物抑制了p31病毒粒子蛋白与艾滋病血清的反应性,这表明它是病毒p31蛋白的一个片段。该肽与所有20名艾滋病患者的血清发生反应,但与所检测的8名正常对照的血清均无反应。这些结果表明该肽可能有助于检测血液样本中的抗HTLV - III抗体。

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