Farias Kleber Juvenal Silva, Machado Paula Renata Lima, de Almeida Junior Renato Ferreira, de Aquino Ana Alice, da Fonseca Benedito Antônio Lopes
Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900, Ribeirão Preto, SP, Brazil; Program of Graduate Studies of Applied Microbiology and Immunology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900, Ribeirão Preto, SP, Brazil; Virology Research Center, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900, Ribeirão Preto, SP, Brazil.
Microbiol Immunol. 2014 Jun;58(6):318-26. doi: 10.1111/1348-0421.12154.
The objective of this study was to investigate the use of chloroquine (CLQ) as an antiviral agent against dengue. Chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal pH, was used in the in vitro treatment of U937 cells infected with dengue virus type 2 (DENV-2). Viral replication was assessed by quantification of virus produced through detection of copy numbers of DENV-2 RNA, plaque assay and indirect immunofluorescence. qRT-PCR and plaque assays were used to quantify the DENV-2 load in infected U937 cells after CLQ treatment. It was found that a dose of 50 μg/mL of CLQ was not toxic to the cells and resulted in significantly less virus production in infected U937 cells than occurred in untreated cells. In the present work, CLQ was effective against DENV-2 replication in U937 cells, and also caused a statistically significant reduction in expression of proinflammatory cytokines. The present study indicates that CLQ may be used to reduce viral yield in U937 cells.
本研究的目的是调查氯喹(CLQ)作为抗登革热病毒剂的用途。氯喹是一种胺酸亲和性药物,已知可通过提高内体pH值来影响细胞内胞吐途径,用于体外治疗感染2型登革热病毒(DENV-2)的U937细胞。通过检测DENV-2 RNA拷贝数、空斑试验和间接免疫荧光来定量产生的病毒,从而评估病毒复制情况。使用qRT-PCR和空斑试验来定量CLQ处理后感染的U937细胞中的DENV-2载量。结果发现,50μg/mL的CLQ剂量对细胞无毒,并且与未处理的细胞相比,感染的U937细胞中产生的病毒显著减少。在本研究中,CLQ对U937细胞中的DENV-2复制有效,并且还导致促炎细胞因子的表达在统计学上显著降低。本研究表明,CLQ可用于降低U937细胞中的病毒产量。