Van Herrewege Yven, Morellato Laurence, Descours Anne, Aerts Laetitia, Michiels Jo, Heyndrickx Leo, Martin Loïc, Vanham Guido
Virology Unit, Department of Microbiology, Institute of Tropical Medicine, Antwerp, Belgium.
J Antimicrob Chemother. 2008 Apr;61(4):818-26. doi: 10.1093/jac/dkn042. Epub 2008 Feb 12.
The antiviral activity of CD4 miniproteins was evaluated as potential HIV microbicides, using relevant in vitro models.
Compounds were tested in a single-cycle HIV-1 pseudovirus assay and against replication competent HIV-1 in co-cultures of monocyte-derived dendritic cells (MO-DC) and CD4+ T cells. Cytotoxic activity was evaluated in an MTT assay.
Monomeric miniproteins (M47 and M48) showed 50% effective concentration (EC(50)) values of 79-105 nM against a subtype B, CCR5 co-receptor-using Ba-L pseudovirus. Higher activity was found for the dimeric miniproteins M48D30, M48D50 and M48D100 (EC(50) between 15 and 30 nM), in contrast to the tetrameric miniproteins M48T30, M48T50 and M48T100 (EC(50) between 107 and 377 nM). The hetero-bivalent miniprotein M48-Hep and miniproteins that targeted the Phe-43 cavity on gp120 (M48-U1, M48-U2 and M48-U3) were highly active, with EC(50) values as low as 2 nM for M48-U1. All miniproteins showed high activity against CCR5 or CXCR4 co-receptor-using subtype B and CRF-01_A/E pseudoviruses. Many early M48-based compounds were much less active against subtype C pseudoviruses, whereas M48-U compounds that targeted the Phe-43 cavity were very active against all pseudoviruses, including subtype C. In MO-DC/CD4+ T cell co-cultures with replication-competent HIV-1 Ba-L, EC(50) values ranged between 13 and 1719 nM depending on the miniprotein, with M48-U1, M48-U2 and M48-U3 again being the most potent. Importantly, the latter compounds completely prevented viral replication by treating the cultures from 2 h before until 24 h after infection, at non-toxic concentrations of 66-6564 nM.
These novel CD4 miniproteins might constitute a promising class of HIV microbicides.
使用相关体外模型评估CD4微型蛋白作为潜在HIV杀微生物剂的抗病毒活性。
在单周期HIV-1假病毒试验中以及在单核细胞衍生的树突状细胞（MO-DC）和CD4+T细胞的共培养物中针对具有复制能力的HIV-1对化合物进行测试。在MTT试验中评估细胞毒性活性。
单体微型蛋白（M47和M48）对使用CCR5共受体的B亚型Ba-L假病毒显示出50%有效浓度（EC(50)）值为79-105 nM。二聚体微型蛋白M48D30、M48D50和M48D100具有更高的活性（EC(50)在15至30 nM之间），而四聚体微型蛋白M48T30、M48T50和M48T100（EC(50)在107至377 nM之间）则不然。异源二价微型蛋白M48-Hep以及靶向gp120上Phe-43腔的微型蛋白（M48-U1、M48-U2和M48-U3）具有高活性，M48-U1的EC(50)值低至2 nM。所有微型蛋白对使用CCR5或CXCR4共受体的B亚型和CRF-01_A/E假病毒均显示出高活性。许多早期基于M48的化合物对C亚型假病毒的活性要低得多，而靶向Phe-43腔的M48-U化合物对包括C亚型在内的所有假病毒都非常活跃。在与具有复制能力的HIV-1 Ba-L的MO-DC/CD4+T细胞共培养物中，EC(50)值根据微型蛋白的不同在13至1719 nM之间，其中M48-U1、M48-U2和M48-U3再次最为有效。重要的是，后一种化合物在66-6564 nM的无毒浓度下，通过在感染前2小时直至感染后24小时处理培养物，完全阻止了病毒复制。
这些新型CD4微型蛋白可能构成一类有前景的HIV杀微生物剂。
