Seesuay Watee, Phanthong Siratcha, Densumite Jaslan, Mahasongkram Kodchakorn, Sookrung Nitat, Chaicumpa Wanpen
Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Center of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Vaccines (Basel). 2021 Aug 12;9(8):893. doi: 10.3390/vaccines9080893.
HIV-1 progeny are released from infected cells as immature particles that are unable to infect new cells. Gag-Pol polyprotein dimerization via the reverse transcriptase connection domain (RTCDs) is pivotal for proper activation of the virus protease (PR protein) in an early event of the progeny virus maturation process. Thus, the RTCD is a potential therapeutic target for a broadly effective anti-HIV agent through impediment of virus maturation. In this study, human single-chain antibodies (HuscFvs) that bound to HIV-1 RTCD were generated using phage display technology. Computerized simulation guided the selection of the transformed -derived HuscFvs that bound to the RTCD dimer interface. The selected HuscFvs were linked molecularly to human-derived-cell-penetrating peptide (CPP) to make them cell-penetrable (i.e., become transbodies). The CPP-HuscFvs/transbodies produced by a selected transformed clone were tested for anti-HIV-1 activity. CPP-HuscFvs of transformed clone 11 (CPP-HuscFv11) that presumptively bound at the RTCD dimer interface effectively reduced reverse transcriptase activity in the newly released virus progeny. Infectiousness of the progeny viruses obtained from CPP-HuscFv11-treated cells were reduced by a similar magnitude to those obtained from protease/reverse transcriptase inhibitor-treated cells, indicating anti-HIV-1 activity of the transbodies. The CPP-HuscFv11/transbodies to HIV-1 RTCD could be an alternative, anti-retroviral agent for long-term HIV-1 treatment.
HIV-1子代以未成熟颗粒的形式从受感染细胞中释放出来，这些颗粒无法感染新细胞。在子代病毒成熟过程的早期事件中，通过逆转录酶连接域（RTCDs）实现的Gag-Pol多蛋白二聚化对于病毒蛋白酶（PR蛋白）的正确激活至关重要。因此，RTCD是通过阻碍病毒成熟来开发广泛有效的抗HIV药物的潜在治疗靶点。在本研究中，利用噬菌体展示技术产生了与HIV-1 RTCD结合的人单链抗体（HuscFvs）。计算机模拟指导了对与RTCD二聚体界面结合的转化衍生HuscFvs的筛选。将所选的HuscFvs与源自人类的细胞穿透肽（CPP）进行分子连接，使其具有细胞穿透性（即成为穿膜抗体）。对由选定的转化克隆产生的CPP-HuscFvs/穿膜抗体进行抗HIV-1活性测试。推测在RTCD二聚体界面结合的转化克隆11的CPP-HuscFvs（CPP-HuscFv11）有效降低了新释放的病毒子代中的逆转录酶活性。从CPP-HuscFv11处理的细胞中获得的子代病毒的感染性降低幅度与从蛋白酶/逆转录酶抑制剂处理的细胞中获得的子代病毒相似，表明穿膜抗体具有抗HIV-1活性。针对HIV-1 RTCD的CPP-HuscFv11/穿膜抗体可能是用于长期治疗HIV-1的另一种抗逆转录病毒药物。
