Identification of MYH9 Key Domain Involved in the Entry of PRRSV Into Permissive Cells

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen that causes huge losses economically to the pig industry worldwide. Previous research suggested that receptor dependence is necessary for PRRSV infection. MYH9 and CD163 are indispensable for PRRSV entry into a porcine alveolar macrophage. In the present study, human MYH9 (hMYH9) and mouse MYH9 (mMYH9), similar to swine MYH9, could also accelerate PRRSV infection in pCD163-mediated cell lines. Knockdown of MYH9 activity using the specific small interfering RNA or inhibitor (blebbistatin) concomitantly decreased PRRSV infection. C-terminal fragment of MYH9 (PRA) proteins from different mammalian species contains a conserved binding domain (aa1676-1791) for PRRSV binding, since the recombinant MYH91676−1791protein could inhibit the PRRSV infection significantly. Furthermore, the specific polyclonal antibody of MYH91676−1791 could block PRRSV infection in host cells. These data strongly supported that MYH9, a very important cofactor, participated in PRRSV entry into target cells, which may facilitate the development of a new therapeutic agent to control PRRSV infection.

Rutin prevents EqHV-8 induced infection and oxidative stress via Nrf2/HO-1 signaling pathway

“…The total RNA of cells samples was extracted by TriQuick Reagent (Solarbio, China), and 1 μg of the total RNA was converted to cDNA using the PrimeScript™ RT Master Mix kit (Takara, Japan) according to the manufacturer’s instructions. The qPCR was performed using a StepOnePlus ® Real-Time PCR applied biosystems (Thermo, USA), the gene expressions of gD, Nrf2, and HO-1 were detected and normalized against those of GADPH using the 2 −ΔΔCt threshold cycle (CT) method as previously described ( Li et al., 2022 ). All primers are listed in Table 1 .…”

“…The siRNAs targeting the Nrf2 and HO-1 genes were synthesized by Ribo Biotechnology Co., Ltd. (Guangdong, China). The siRNA knockdown assay was performed as previously described ( Li et al., 2022 ). In brief, the RK-13 cells were transfected with these siRNAs according to the Lipo 6000 protocol (Beyotime, China), respectively, and non-targeting siRNA served as a negative control.…”

Hyperoside inhibits EHV-8 infection via alleviating oxidative stress and IFN production through activating JNK/Keap1/Nrf2/HO-1 signaling pathways

“…The CCK-8 was obtained from Beyotime Biotechnology (Nanjing, China), and it was used to detect the cytotoxicity of hyperoside in RK-13, MDBK, and NBL-6 cells. Cells in 96-well plates were incubated with or without hyperoside for 24 h. Then, 100 µL MEM containing 10% CCK-8 reagent was added to each well, and the cells were incubated at 37°C for another 2 h. The cell viability was measured at an absorbance of 450 nm using a Spectramax Absorbance Reader (USA) and analyzed using GraphPad Prism 8.0 as previously described ( 50 ).…”

The Antiviral Activity of Caprylic Monoglyceride against Porcine Reproductive and Respiratory Syndrome Virus In Vitro and In Vivo

“…Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which was first identified in the United States in 1987 and is an economically impactful disease in the global pig industry [ 1 , 2 ]. To date, PRRSV is known specifically to infect pigs, and there are no data to suggest that other species are susceptible to this pathogen [ 3 ]. PRRSV replicates primarily in porcine alveolar macrophages, and the clinical signs of PRRSV infection vary among pigs at different ages.…”