Restriction of porcine reproductive and respiratory syndrome virus replication by galectin-1
VETERINARY MICROBIOLOGY
Authors: Li, Liwei; Zhao, Kuan; Gao, Fei; Jiang, Yifeng; Shan, Tongling; Tong, Wu; Zheng, Hao; Yu, Lingxue; Li, Guoxin; Ma, Zhiyong; Tong, Guangzhi
Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) causes great economic losses to the swine industry globally; however, effective control measures for this virus are limited. Here, we screened a porcine alveolar macrophage (PAM) cDNA library with a yeast two-hybrid system to reveal that galectin-1 (Gal-1), an endogenous innate immune protein encoded by LGALS1, interacts with nonstructural protein 11 (Nsp11) of PRRSV. Western blotting and viral titer assays indicated that Gal-1 overexpression suppressed replication in multiple PRRSV strains (P < 0.001), whereas Gal-1 knockdown or knockout increased viral titer and nucleocapsid protein expression. The Gal-l-specific anti-PRRSV effect was associated with the endoribonuclease domain of Nsp11 through inactivation of interferon-antagonist function and stimulation of interferon-stimulated gene expression. Additionally, Gal-1 interacted with PRRSV E protein but not with PRRSV glycoproteins, and recombinant Gal-1 treatment inhibited PRRSV in PAMs and MARC-145 cells. Furthermore, Gal-1 inhibited replication in multiple viruses, including equine arteritis virus, porcine epidemic diarrhea virus, pseudorabies virus, Japanese encephalitis virus, and classical swine fever virus, suggesting its potential broad application for antiviral strategies. Our findings provide insight into the important role of Gal-1 in PRRSV pathogenesis and its potential use as a novel therapeutic target against PRRSV infection.
Differential detection of porcine reproductive and respiratory syndrome virus genotypes by a fluorescence melting curve analysis using peptide nucleic acid probe-mediated one-step real-time RT-PCR
JOURNAL OF VIROLOGICAL METHODS
Authors: Park, Ji-Young; Kim, Seong-Hee; Lee, Kyoung-Ki; Kim, Yeon-Hee; Moon, Bo-Yeon; So, ByungJae; Park, Choi-Kyu
Abstract
Peptide nucleic acids (PNAs), artificially synthesized DNA analogues, hybridize strongly with DNA and are useful for fluorescence melting curve analyses (FMCA) based on the thermal denaturation of the probe-target duplex. In this study, we developed a PNA-based one-step real-time RT-PCR assay for the differential and qualitative detection of the porcine reproductive and respiratory syndrome virus genotypes PRRSV1 and PRRSV2. The specificity of the assay was analyzed in silico using previously reported primers and probes and was subsequently verified using Korean PRRSV panels and clinical samples. Seven clinical samples showing low curves with high Ct values were confirmed as negative by FMCA. The sensitivities of one-step real-time PCR for PRRSV1 and PRRSV2 were 15 and 11 copies, respectively, and the results were in 100% agreement with those of conventional RT-PCR combined with nested PCR using clinical samples. Therefore, the assay is highly specific for the detection of current PRRSV1 and PRRSV2 without non-specific amplification by FMCA.