Espinoza Daniela, Laporte Daniel, Martínez Fabián, Sandino Ana María, Valdés Natalia, Moenne Alejandra, Imarai Mónica
Laboratorio de Inmunología, Centro de Biotecnología Acuícola (CBA), Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile., Alameda, 3363 Santiago, Chile; Laboratory of Marine Biotechnology, Faculty of Chemistry and Biology, University of Santiago de Chile, Biology, Alameda, 3363 Santiago, Chile.
Laboratory of Plant Physiology and Molecular Biology, Instituto de Ciencias Biomédicas, Universidad Autónoma de Chile, Talca 3467987, Chile.
Int J Biol Macromol. 2024 Dec;282(Pt 2):136875. doi: 10.1016/j.ijbiomac.2024.136875. Epub 2024 Oct 23.
Sulfated polysaccharide lambda carrageenan (λ-CGN) was evaluated for its antiviral effect against IPNV using the in vitro infection model of CHSE-214 salmonid cells. A plaque reduction lysis assay revealed that λ-CGN has an IC of 0.9 μg⋅mL, CC > 128 μg⋅mL and a Selectivity Index (SI) > 142. In comparison, iota, kappa carrageenans and lambda oligo-carrageenan (λ-OC) were less effective than λ-CGN against IPNV. λ-CGN showed no virucidal activity when applied directly to viral particles. Time of addition experiments showed that pre-treatment, co-treatment, and post-treatment with λ-CGN significantly reduced viral RNA copies in the cell supernatant. Additionally, a decrease in intracellular viral RNA was observed with pre-treatment and post-treatment, indicating an impact on different stages of viral replication. Polyacrylamide gel electrophoresis of IPNV genomic RNA in presence of λ-CGN showed a reduction in the level of viral genomic RNA. Confocal microscopy confirmed the intracellular localization of λ-CGN, suggesting that λ-CGN may inhibit the synthesis of IPNV genomic RNA. Moreover, cells pre-treated with λ-CGN showed an increased expression of innate immunity genes CXCL11, IL1β, IFNa, and IRF3. These findings highlight the need for further research to confirm the in vivo pharmacological potential of λ-CGN as a new antiviral agent in salmonids.
使用CHSE - 214鲑鱼细胞的体外感染模型,评估了硫酸化多糖λ-角叉菜胶(λ-CGN)对传染性胰腺坏死病毒(IPNV)的抗病毒作用。噬斑减少裂解试验表明,λ-CGN的半数抑制浓度(IC)为0.9μg·mL,细胞毒性浓度(CC)>128μg·mL,选择性指数(SI)>142。相比之下,ι-角叉菜胶、κ-角叉菜胶和λ-寡聚角叉菜胶(λ-OC)对IPNV的抗病毒效果不如λ-CGN。将λ-CGN直接应用于病毒颗粒时未显示出杀病毒活性。添加时间实验表明,用λ-CGN进行预处理、共处理和后处理可显著降低细胞上清液中的病毒RNA拷贝数。此外,预处理和后处理均观察到细胞内病毒RNA减少,表明对病毒复制的不同阶段有影响。在λ-CGN存在下对IPNV基因组RNA进行聚丙烯酰胺凝胶电泳显示病毒基因组RNA水平降低。共聚焦显微镜证实了λ-CGN的细胞内定位,表明λ-CGN可能抑制IPNV基因组RNA的合成。此外,用λ-CGN预处理的细胞显示先天免疫基因CXCL11、IL1β、IFNa和IRF3的表达增加。这些发现突出了进一步研究以确认λ-CGN作为鲑鱼中新抗病毒剂的体内药理潜力的必要性。