Ponomarenko Natalia A, Vorobiev Ivan I, Alexandrova Elena S, Reshetnyak Andrew V, Telegin Georgy B, Khaidukov Sergey V, Avalle Bérangère, Karavanov Alexander, Morse Herbert C, Thomas Daniel, Friboulet Alain, Gabibov Alexander G
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, RAS, 16/10, Miklukho-Maklaya str., Moscow 117871, Russia.
Biochemistry. 2006 Jan 10;45(1):324-30. doi: 10.1021/bi050675k.
We have induced a polyclonal IgG that degrades the HIV-1 surface antigen, glycoprotein gp120, by taking advantage of the susceptibility of SJL mice to a peptide-induced autoimmune disorder, experimental autoimmune encephalomyelitis (EAE). Specific pathogen-free SJL mice were immunized with structural fragments of gp120, fused in-frame with encephalitogenic peptide MBP(85-101). It has resulted in a pronounced disease-associated immune response against antigens. A dramatic increase of gp120 degradation level by purified polyclonal IgG from immunized versus nonimmunized mice has been demonstrated by a newly developed fluorescence-based assay. This activity was inhibited by anti-mouse immunoglobulin antibodies as well as by Ser- and His-reactive covalent inhibitors. A dominant proteolysis site in recombinant gp120 incubated with purified polyclonal IgG from immunized mice was shown by SDS-PAGE. The SELDI-based mass spectrometry revealed that these antibodies exhibited significant specificity toward the Pro484-Leu485 peptide bond. The sequence surrounding this site is present in nearly half of the HIV-I variants. This novel strategy can be generalized for creating a catalytic vaccine against viral pathogens.
我们利用SJL小鼠对肽诱导的自身免疫性疾病——实验性自身免疫性脑脊髓炎（EAE）的易感性，诱导出了一种能降解HIV-1表面抗原糖蛋白gp120的多克隆IgG。将无特定病原体的SJL小鼠用gp120的结构片段进行免疫，该片段与致脑炎性肽MBP(85-101)进行读框融合。这引发了针对抗原的明显的疾病相关免疫反应。通过一种新开发的基于荧光的检测方法，已证明免疫小鼠与未免疫小鼠的纯化多克隆IgG对gp120的降解水平有显著提高。这种活性受到抗小鼠免疫球蛋白抗体以及丝氨酸和组氨酸反应性共价抑制剂的抑制。SDS-PAGE显示了与免疫小鼠的纯化多克隆IgG一起孵育的重组gp120中的一个主要蛋白水解位点。基于表面增强激光解吸电离飞行时间质谱（SELDI）的质谱分析表明，这些抗体对Pro484-Leu485肽键具有显著特异性。该位点周围的序列存在于近一半的HIV-1变体中。这种新策略可推广用于创建针对病毒病原体的催化疫苗。
