De Santis R, Anastasi A M, Marcolini S, Valesini G, Pezzella M, Vonesch N, Sturchio E, Mele A
Department of Biotechnology, Menarini Ricerche Sud, Pomezia, Rome, Italy.
AIDS Res Hum Retroviruses. 1991 Mar;7(3):315-21. doi: 10.1089/aid.1991.7.315.
Monoclonal antibodies have been generated against a synthetic peptide of the nef protein of human immunodeficiency virus type 1 (HIV-1) in order to further characterize the biochemical and functional nature of this protein and its role in the control of HIV-1 transcriptional regulation. Earlier studies indicated nef to be a negative regulatory factor for viral transcription, whereas more recent studies report evidence against this original hypothesis. Nef is a protein of 206 amino acids of approximately 27 kD in most HIV-1 isolates; however, in some other isolates a truncated form of 124 amino acids has been described. A peptide sequence of six amino acids, corresponding to a region of the nef protein exhibiting high-sequence homology to thymosin alpha 1 protein, has been synthesized by Merrifield solid-phase methodology. This peptide is coded by a sequence located upstream to the stop codon described in some HIV-1 isolates and then is maintained in both complete and truncated forms of the nef protein. F14.11 is a nef peptide-specific monoclonal antibody (IgG2a/k) exhibiting the ability to recognize natural nef protein in either radioimmunoassay, radioimmunoprecipitation assay, or immunocytochemical analysis. Since F14.11 is able to identify nef protein in the cytoplasm of lymphocytes from HIV-infected seronegative subjects it may prove useful in monitoring the expression of nef during the silent HIV-1 infection.
已针对人类免疫缺陷病毒1型（HIV-1）的nef蛋白的合成肽产生了单克隆抗体，以便进一步表征该蛋白的生化和功能性质及其在HIV-1转录调控控制中的作用。早期研究表明nef是病毒转录的负调控因子，而最近的研究报告了反对这一原始假设的证据。在大多数HIV-1分离株中，Nef是一种由206个氨基酸组成、分子量约为27kD的蛋白质；然而，在其他一些分离株中，已描述了一种124个氨基酸的截短形式。通过Merrifield固相方法合成了一段由六个氨基酸组成的肽序列，该序列对应于nef蛋白中与胸腺素α1蛋白具有高度序列同源性的区域。该肽由一些HIV-1分离株中所述终止密码子上游的序列编码，然后在nef蛋白的完整和截短形式中均保留。F14.11是一种nef肽特异性单克隆抗体（IgG2a/k），在放射免疫测定、放射免疫沉淀测定或免疫细胞化学分析中均表现出识别天然nef蛋白的能力。由于F14.11能够识别来自HIV感染血清阴性受试者淋巴细胞胞质中的nef蛋白，因此它可能在监测无症状HIV-1感染期间nef的表达方面证明是有用的。
