Research on Replication of BVDV Through Exosome Pathway Mediated by Host Protein SPCS1
ZHANG Cheng-Yuan1, WEI Yu-Rong2, CHEN Jun-Zhen1, YANG Li1, LI Ze-Yu1, LI Zi-Qian1, FU Qiang1,*, SHI Hui-Jun1,*
1 College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China; 2 Xinjiang Key Laboratory of Animal Disease Research, Institute of Veterinary Medicine, Xinjiang Academy of Animal Science, Urumqi 830052, China
Abstract Bovine viral diarrhea (BVD) is an infectious disease of cattle worldwide caused by Bovine viral diarrhea virus (BVDV), which has brought huge economic losses to animal husbandry. The previous study found that knocked down the expression of signal peptidase complex subunit 1 (SPCS1) in bovine kidney cells (Madin-Darby Bovine Kidney, MDBK) could inhibit the replication of BVDV effectively. In order to explore its molecular mechanism, this study collected exosomes from the supernatant of SPCS1 knockdown (KD) cells infected with BVDV, and transmission electron microscopy, Western blot and Nanoparticle Tracking Analysis were used to characterize exosomes. qPCR was used to analysis BVDV 5' untranslated region (UTR) RNA levels in the exosomes. The exosomes were infected with MDBK; BVDV 5'UTR RNA levels were detected by qPCR; The cytopathic effect (CPE) was observed by inverted microscope, and the change of virus titer was determined by Reed-Muench method. The exosomes were observed as disk-like vesicles under transmission electron microscopy, Western blot detected exosomes, the volume characteristic protein TSG101 showed a peak value at about 100 nm, indicating exosomes were successfully extracted. Western blot showed that the expression of exosome TSG101 protein in SPCS1 KD cells infected with BVDV was significantly decreased compared with the control cells treated with empty vector (Scramble cells)(P<0.001). qPCR detection showed that the level of BVDV 5'UTR mRNA in exosomes was significantly decreased (P<0.001). BVDV 5'UTR RNA levels of SPCS1 KD cell derived exosomes infected with MDBK were significantly lower than those infected with Scramble cell (P<0.001), the CPE phenomenon was delayed and weakened, and the viral titer of the progeny was significantly decreased (P< 0.001). The results showed that knockdown SPCS1 affected the replication of BVDV through exosome pathway. The research results provide a new antiviral target and theoretical basis for the prevention and control of BVDV.
ZHANG Cheng-Yuan,WEI Yu-Rong,CHEN Jun-Zhen, et al. Research on Replication of BVDV Through Exosome Pathway Mediated by Host Protein SPCS1[J]. 农业生物技术学报, 2024, 32(11): 2662-2669.
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