Malley S D, Grange J M, Hamedi-Sangsari F, Vila J R
Compagnie de Développement Aguettant Société Anonyme, Lyon, France.
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11017-21. doi: 10.1073/pnas.91.23.11017.
The cellular models generally used in the in vitro evaluation of anti-human immunodeficiency virus compounds are dividing cells. A model constituted by resting lymphocytes may more accurately reflect a drug's future efficacy in humans, since viral DNA synthesis is known to take place in quiescent cells, creating a reservoir of infected cells awaiting activation to complete their viral replication cycle and to produce infectious virions. We report here the activity of 3'-azido-3'-deoxythymidine, 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, and two hydroxamates, D-aspartic acid beta-hydroxamate and hydroxycarbamate (hydroxyurea), alone and in various combinations, in an in vitro model based on resting lymphocytes. In our model, resting peripheral blood lymphocytes were infected with human immunodeficiency virus type 1 and treated with drugs for 7 days, at which time drugs were removed and the cells were activated by phytohemagglutinin. We show that under these conditions 3'-azido-3'-deoxythymidine, 2',3'-dideoxyinosine, and 2',3'-dideoxycytidine, alone or in combination, neither fully inhibit viral production nor protect lymphocytes from the cytopathic effect of viral replication, at concentrations corresponding to the peak plasma levels observed in a typical treatment schedule in humans. In contrast, we report the synergistic effect of treatment by each hydroxamate with 2',3'-dideoxyinosine of infected resting lymphocytes, resulting in the total suppression of viral production, total protection against the cytopathic effect induced by viral replication, and no effect on the ability of the cells to replicate in this cell culture system.
在体外评估抗人类免疫缺陷病毒化合物时通常使用的细胞模型是分裂细胞。由静息淋巴细胞构成的模型可能更准确地反映药物在人体中的未来疗效,因为已知病毒DNA合成发生在静止细胞中,从而形成一个感染细胞库,等待激活以完成其病毒复制周期并产生感染性病毒粒子。我们在此报告3'-叠氮基-3'-脱氧胸苷、2',3'-双脱氧肌苷、2',3'-双脱氧胞苷以及两种异羟肟酸(D-天冬氨酸β-异羟肟酸和羟基脲)单独及以各种组合形式在基于静息淋巴细胞的体外模型中的活性。在我们的模型中,静息外周血淋巴细胞感染1型人类免疫缺陷病毒并用药物处理7天,此时去除药物,然后用植物血凝素激活细胞。我们表明,在这些条件下,3'-叠氮基-3'-脱氧胸苷、2',3'-双脱氧肌苷和2',3'-双脱氧胞苷单独或联合使用时,在对应于人类典型治疗方案中观察到的血浆峰值水平的浓度下,既不能完全抑制病毒产生,也不能保护淋巴细胞免受病毒复制的细胞病变效应。相比之下,我们报告了每种异羟肟酸与感染的静息淋巴细胞的2',3'-双脱氧肌苷联合治疗的协同作用,导致病毒产生完全受到抑制,完全保护免受病毒复制诱导的细胞病变效应,并且对细胞在该细胞培养系统中的复制能力没有影响。