Palma-Ocampo Helen K, Flores-Alonso Juan C, Vallejo-Ruiz Verónica, Reyes-Leyva Julio, Flores-Mendoza Lilian, Herrera-Camacho Irma, Rosas-Murrieta Nora H, Santos-López Gerardo
Laboratorio de Biología Molecular y Virología, Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Metepec, Puebla, México.
Posgrado en Ciencias Químicas, Benemérita Universidad Autónoma de Puebla, Puebla, México.
Virol J. 2015 Sep 28;12:150. doi: 10.1186/s12985-015-0383-4.
In viral disease, infection is controlled at the cellular level by type I interferon (IFN-I), but dengue virus (DENV) has the ability to inhibit this response. Type III interferon, also known as lambda IFN (IFN-III or IFN-λ), is a complementary pathway to the antiviral response by IFN-I. This work analyzed the IFN-λ (IFN-III) mediated antiviral response against DENV serotype 2 (DENV-2) infection.
Dengue fever patients were sampled to determine their IFN-λ levels by ELISA. To study the IFN-λ response during DENV infection we selected the epithelial cell line C33-A, and we demonstrated that it is permissive to DENV-2 infection. The effect of IFN-λ on virus replication was determined in these cells, in parallel to the expression of IFN-stimulated genes (ISGs), and Suppressor of Cytokine Signaling (SOCS), genes measured by RT-qPCR.
We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors. IFN-λ inhibited DENV-2 replication in a dose-dependent manner in vitro. The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression. Presence of IFN-negative regulators, SOCS1 and SOCS3, during DENV-2 infection was associated with reduced IFN-λ1 expression.
Evidence described here suggests that IFN-λ is a good candidate inhibitor of viral replication in dengue infection. Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease. Furthermore, we report a novel epithelial model to study dengue infection in vitro.
在病毒疾病中,I型干扰素(IFN-I)在细胞水平上控制感染,但登革热病毒(DENV)有能力抑制这种反应。III型干扰素,也称为λ干扰素(IFN-III或IFN-λ),是IFN-I介导的抗病毒反应的补充途径。本研究分析了IFN-λ(IFN-III)介导的针对登革热病毒2型(DENV-2)感染的抗病毒反应。
采集登革热患者样本,通过酶联免疫吸附测定(ELISA)确定其IFN-λ水平。为了研究DENV感染期间的IFN-λ反应,我们选择了上皮细胞系C33-A,并证明它对DENV-2感染敏感。在这些细胞中确定IFN-λ对病毒复制的影响,同时通过逆转录定量聚合酶链反应(RT-qPCR)检测IFN刺激基因(ISG)和细胞因子信号抑制因子(SOCS)基因的表达。
我们发现,与健康献血者相比,登革热患者血清中的IFN-λ增加了(约1.8倍)。IFN-λ在体外以剂量依赖的方式抑制DENV-2复制。病毒滴度的降低与ISG mRNA水平(MX1和OAS1)的增加相对应,在ISG表达峰值时抑制作用最强。在DENV-2感染期间,IFN负调节因子SOCS1和SOCS3的存在与IFN-λ1表达的降低有关。
此处所述证据表明,IFN-λ是登革热感染中病毒复制的良好候选抑制剂。DENV与IFN-λ之间细胞和机体相互作用的机制需要进一步研究,因为它们可能为治疗该疾病的策略提供见解。此外,我们报告了一种用于体外研究登革热感染的新型上皮模型。