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以及对与G4-PAMAM树枝状大分子复合的gp120-HIV衍生肽的研究。

and studies of gp120-HIV-derived peptides in complex with G4-PAMAM dendrimers.

作者信息

Rolando Alberto Rodríguez-Fonseca, Martiniano Bello, Saúl Rojas-Hernández, Jazmín García-Machorro, Mara Gutiérrez-Sánchez, Alan Rubén Estrada-Pérez, Manuel Jonathan Fragoso-Vázquez, Juan Vicente Méndez-Méndez, José Correa-Basurto

机构信息

Laboratorio de Diseño y Desarrollo de Nuevos Fármacos e Innovación Biotécnológica (Laboratory for the Design and Development of New Drugs and Biotechnological Innovation), Escuela Superior de Medicina, Instituto Politécnico Nacional Plan de San Luis y Díaz Mirón Ciudad de México 11340 Mexico

Laboratorio de Inmunología Molecular, Escuela Superior de Medicina, Instituto Politécnico Nacional Plan de San Luis y Díaz Mirón, Ciudad de México 11340 Mexico.

出版信息

RSC Adv. 2020 May 27;10(35):20414-20426. doi: 10.1039/d0ra00840k.

Abstract

Novel synthetic vaccines as immunotherapy approaches for HIV are interesting strategies that imply big challenges as they increase the poor immunogenic properties of peptide epitopes and their structural damage from the physiological environment. In this work, we used fourth-generation polyamidoamine dendrimers (G4-PAMAM) to increase the immunoglobulin responses () induced by two peptide epitopes (pPGT122: DIIGDIRQAH and pVRC03: DGGANNTSNETFR), both recognized by broadly neutralizing antibodies (bNAb) on gp120-HIV type 1. pPGT122 and pVRC03 were identified on the gp120 surface recognition by bNAb by using X-ray diffraction-derived structures obtained from the Protein Data Bank. pPGT122 and pVRC03 were coupled to the G4-PAMAM molecule by ligand diffusion using molecular dynamics (LDMDS) simulations and their energetic values were calculated by using the MMGBSA approach. Additionally, docking and MD simulations showed the affinity of pPGT122 and pVRC03 for MHC-I/II. G4-PAMAM-peptide complexes were chemically characterized through MALDI-TOF-MS, LC-ESI-QTOF-MS, atomic force microscopy (AFM) and H NMR spectroscopy. Then, the G4-PAMAM-peptide complexes were assayed by intranasal administration in female BALB/cJ mouse groups, showing that both peptides were immunogenic systemically and in the mucous membrane (in nasal and vaginal washes) increase in IgG and IgA, respectively. This demonstrated that G4-PAMAM can be used as a nanocarrier for immunogenic peptides.

摘要

新型合成疫苗作为治疗HIV的免疫疗法是很有趣的策略,但也意味着巨大挑战,因为它们会增加肽表位的免疫原性差的特性以及其在生理环境中的结构损伤。在这项工作中,我们使用第四代聚酰胺胺树枝状大分子(G4-PAMAM)来增强两种肽表位(pPGT122:DIIGDIRQAH和pVRC03:DGGANNTSNETFR)诱导的免疫球蛋白反应,这两种肽表位均能被1型HIV gp120上的广泛中和抗体(bNAb)识别。通过使用从蛋白质数据库获得的X射线衍射衍生结构,在gp120表面上鉴定出pPGT122和pVRC03可被bNAb识别。通过使用分子动力学(LDMDS)模拟通过配体扩散将pPGT122和pVRC03与G4-PAMAM分子偶联,并使用MMGBSA方法计算其能量值。此外,对接和MD模拟显示了pPGT122和pVRC03对MHC-I/II的亲和力。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)、液相色谱-电喷雾-四极杆飞行时间质谱(LC-ESI-QTOF-MS)、原子力显微镜(AFM)和核磁共振氢谱(H NMR)对G4-PAMAM-肽复合物进行化学表征。然后,通过鼻内给药对雌性BALB/cJ小鼠组进行G4-PAMAM-肽复合物检测,结果表明两种肽在全身和黏膜(鼻腔和阴道冲洗液中)均具有免疫原性,分别使IgG和IgA增加。这表明G4-PAMAM可作为免疫原性肽的纳米载体。

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