ADS-J1对SEVI增强HIV-1初始传播病毒及其慢性控制病毒感染的拮抗作用及机制

摘要

Objective To investigate the effect of semen-derived enhancer of virus infection (SEVI) on the infection of transmitted/founder (TF) HIV-1 and its matched chronic control (CC) viruses and the antagonism of ADS-J1 on SEVI-mediated enhancement of TF and CC virus infection in vitro. Methods PAP248-286 self-assembling into SEVI amyloid fibrils was validated by ThT assay. We generated the virus stocks of TF and CC virus pair. TZM-bl cells were infected with the mixture of SEVI and TF or CC viruses for 72 h. Luciferase activity was used to observe the enhancement of SEVI. SEVI was treated with different concentrations of ADS-J1 and incubated with TF or CC viruses. TZM-bl cells were then infected with the mixture and luciferase activity was detected 72 h after infection to analyze the antagonism of ADS-J1 on the enhancing effect of SEVI. ADS-J1 was also incubated with TF and CC viruses directly and TZM-bl cells were infected for 72 h to evaluate the antiviral effect using luciferase assay. SEVI was treated with ADS-J1 and Zeta potential was determined to explore the antagonistic mechanism of ADS-J1. Results ThT assay showed that PAP248-286 was capable of self-assembly into SEVI amyloid fibrils. SEVI significantly accelerated TF and CC viruses infection (P<0.05), and ADS-J1 not only significantly antagonized the enhancement of SEVI (P<0.05) but also directly inhibited the infection of TF and CC viruses (P<0.05). ADS-J1 neutralized the positive charge of SEVI in a dose-dependent manner. Conclusions SEVI promotes the infection of TF and CC strains, and ADS-J1 antagonizes SEVI-mediated enhancement of TF and CC viruses by neutralizing the positive charge of SEVI.%目的 探讨精液源性病毒增强因子(SEVI)促进HIV-1初始传播(TF)病毒及其慢性控制(CC)病毒感染的情况,及ADS-J1拮抗SEVI增强病毒感染的作用机制.方法 硫磺素T(ThT)实验验证PAP248-286能自组装成SEVI淀粉样纤维;扩增1对TF和CC感染性克隆病毒,SEVI分别与TF、CC病毒混合后感染TZM-bl细胞,72 h后测定荧光素酶活性,评价SEVI增强病毒感染的倍数;用不同浓度的ADS-J1处理SEVI,再分别与TF、CC病毒混合后感染TZM-bl细胞,72 h后测定荧光素酶活性,考察ADS-J1对SEVI增强TF和CC病毒感染的拮抗作用;接着用ADS-J1和病毒混合后感染TZM-bl细胞,72 h后测定荧光素酶活性,验证ADS-J1直接的抗病毒作用.最后用不同浓度的ADS-J1处理SEVI,检测其Zeta电位,初步探索ADS-J1拮抗SEVI增强TF和CC病毒感染的作用机制.结果 ThT实验结果表明PAP248-286能自组装成SEVI淀粉样纤维;SEVI可显著促进TF和CC病毒的感染(P<0.05),ADS-J1不仅能显著拮抗SEVI增强TF和CC感染(P<0.05)的作用,还能直接抑制TF和CC感染靶细胞(P<0.05);ADS-J1能浓度依赖性地中和SEVI所带的正电荷.结论 SEVI能促进TF和CC病毒的感染,ADS-J1可能通过中和SEVI表面的正电荷来拮抗SEVI增强TF和CC的感染作用.