Anti-viral activity of amphotericin B methyl ester: inhibition of HTLV-III replication in cell culture.
Anti-viral activity of amphotericin B methyl ester: inhibition of HTLV-III replication in cell culture.
复制标题
两性霉素 B 甲酯的抗病毒活性：抑制细胞培养中的 HTLV-III 复制。
DOI：
10.1016/0006-2952(86)90037-7
复制
发表时间：
1986
影响因子：
5.8
通讯作者：
P. Sarin
中科院分区：
文献类型：
--
作者：
C. Schaffner;O. Plescia;D. Pontani;D. Sun;A. Thornton;R. C. Pandey;P. Sarin
材料与方法两性霉素B和杀念菌素得自Xechem，Inc.，伊利诺伊州梅尔罗斯公园将它们溶解在0.01%二甲基甲酰胺（DMF）中以制备浓度范围为0.1至100 μ g/ml的溶液。如Mechlinksi和Schaffner [6]所述制备作为抗坏血酸盐的阿替西霉素B甲酯（AME）。AME盐是水溶性的。因此AME抗坏血酸盐作为水溶液加入到病毒感染的H9细胞培养物中。HTLV-III病毒。从培养的HTLVIII感染的H9细胞制备储备病毒。收获细胞，并通过在蔗糖梯度上两次条带化从条件培养基中分离病毒。收集以1.16至1.19 μ g/ml的密度沉降的病毒带，通过离心沉淀，并用TNE（10 mM Tris·HCl，pH 7.5）重悬。0.1 M NaCl，0.001 M EDTA），以2 X 10 - 12病毒颗粒/ml。H9细胞。这是允许HTLV-III的转化的人淋巴细胞系[1]。使用含有RPMI-1640的培养基（补充有20%胎牛血清、2%谷氨酰胺和1%庆大霉素），通过连续传代在培养物中维持。出于测试目的，在对数期收获细胞，并且每ml测试培养物添加5x 105个细胞。
Materials and methodsTest drugs. Amphotericin B and candicidin were obtained from Xechem, Inc., Melrose Park, IL. They were dissolved in 0.01% dimethylformamide(DMF) to prepare solutions ranging in concentration from 0.1 to 100 pg/ml. The amphotericin B methyl ester (AME), as the ascorbic acid salt, was prepared as described by Mechlinksi and Schaffner [6]. AME salts are water-soluble. in contrast to amphotericin B, so that AME ascorbate was added to cultures of virus-infected H9 cells as an aqueous solution. HTLV-III virus. Stock virus was prepared from HTLVIII infected H9 cells in culture. Cells were harvested and the virus was isolated from the conditioned medium by twice banding on sucrose gradients. The virus band sedimenting at a density of 1.16 to 1.19 g/ml was collected, pelleted by centrifugation, and resuspended by TNE (1OmM Tris’ HCI, pH 7.5. 0.1 M NaCI, 0.001 M EDTA) at 2 X 10” virus particles/ml. H9 cells. This is a transformed human lymphocyte cell line that is permissive for HTLV-III [l]. It is maintained by serial passage in culture, using medium containing RPMI-1640 supplemented with 20% fetal bovine serum, 2% glutamine and 1% gentamicin. For test purposes, cells were harvested in log phase, and 5 x lo5 cells were added per ml of test culture.