PMID- 34149651 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20210622 IS - 1664-302X (Print) IS - 1664-302X (Electronic) IS - 1664-302X (Linking) VI - 12 DP - 2021 TI - RIPK3-Dependent Necroptosis Limits PRV Replication in PK-15 Cells. PG - 664353 LID - 10.3389/fmicb.2021.664353 [doi] LID - 664353 AB - Pigs infected by pseudorabies virus (PRV) display necrotic pathology in multiple organs. The mechanism by which PRV induces cell death is still unclear. Recently, necroptosis was identified as a programmed process dependent on the receptor interacting protein kinase 3 (RIPK3) and mixed lineage kinase-like protein (MLKL). In this study, we demonstrated that PRV induced RIPK3-dependent necroptosis in PK-15 cells. The data showed that PRV infection caused cell death with Propidium Iodide (PI)-positive staining. Transmission electron microscopy analysis indicated plasma membrane disruption in PRV-infected cells. A pan-caspase inhibitor did not prevent PRV-induced necrotic cell death. Western blot analysis indicated that caspase-3 and caspase-8 were not cleaved during PRV infection. Although the transcription of tumor necrosis factor-alpha (TNF-alpha) was increased by PRV infection, RIPK1 was shown to be not involved in PRV-induced necrotic cell death by use of its specific inhibitor. Further experiments indicated that the phosphorylation of RIPK3 and MLKL was upregulated in PRV-infected cells. Stable shRNA knockdown of RIPK3 or MLKL had a recovery effect on PRV-induced necrotic cell death. Meanwhile, viral titers were enhanced in RIPK3 and MLKL knockdown cells. Hence, we concluded that initiation of necroptosis in host cells plays a limiting role in PRV infection. Considering that necroptosis is an inflammatory form of programmed cell death, our data may be beneficial for understanding the necrotic pathology of pigs infected by PRV. CI - Copyright (c) 2021 Gou, Bian, Cai, Chu, Song, Li, Jiang, Zhang, Yang and Li. FAU - Gou, Hongchao AU - Gou H AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Bian, Zhibiao AU - Bian Z AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Cai, Rujian AU - Cai R AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Chu, Pinpin AU - Chu P AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Song, Shuai AU - Song S AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Li, Yan AU - Li Y AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Jiang, Zhiyong AU - Jiang Z AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Zhang, Kunli AU - Zhang K AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Yang, Dongxia AU - Yang D AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. FAU - Li, Chunling AU - Li C AD - Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China. AD - Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China. AD - Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China. LA - eng PT - Journal Article DEP - 20210604 PL - Switzerland TA - Front Microbiol JT - Frontiers in microbiology JID - 101548977 PMC - PMC8211757 OTO - NOTNLM OT - PK-15 cell OT - mixed lineage kinase-like protein (MLKL) OT - necroptosis OT - pseudorabies virus (PRV) OT - receptor interacting protein kinase 3 (RIPK3) COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2021/06/22 06:00 MHDA- 2021/06/22 06:01 PMCR- 2021/06/04 CRDT- 2021/06/21 05:51 PHST- 2021/02/05 00:00 [received] PHST- 2021/04/21 00:00 [accepted] PHST- 2021/06/21 05:51 [entrez] PHST- 2021/06/22 06:00 [pubmed] PHST- 2021/06/22 06:01 [medline] PHST- 2021/06/04 00:00 [pmc-release] AID - 10.3389/fmicb.2021.664353 [doi] PST - epublish SO - Front Microbiol. 2021 Jun 4;12:664353. doi: 10.3389/fmicb.2021.664353. eCollection 2021.