PMID- 26573719 OWN - NLM STAT- MEDLINE DCOM- 20160629 LR - 20240324 IS - 1743-422X (Electronic) IS - 1743-422X (Linking) VI - 12 DP - 2015 Nov 14 TI - Rescue and evaluation of a recombinant PRRSV expressing porcine Interleukin-4. PG - 185 LID - 10.1186/s12985-015-0380-7 [doi] LID - 185 AB - BACKGROUND: The current vaccines for porcine reproductive and respiratory syndrome virus (PRRSV) have failed to provide broad protection against infection by various strains of PRRSV. Porcine Interleukin-4 (pIL-4) plays an important role in the regulation of the immune response and has been used previously as an immunological adjuvant. The objective of this study was to construct a recombinant PRRSV expressing pIL-4 and to evaluate the immune response of the recombinant virus in piglets. METHODS: The pIL-4 gene was inserted in the PRRSV (CH-1R strain) infectious clone by overlap PCR. Indirect immunofluorescence assay (IFA) and Western blotting were used to confirm the recombinant virus. The stability of the recombinant virus was assessed by DNA sequencing and IFA after 15 passages in vitro. Recombinant virus was injected into pigs and efficacy of immune protection was evaluated in comparison with the parental virus. RESULTS: The recombinant virus (CH-1R/pIL-4) was successfully rescued and shown to have similar growth kinetics as the parental virus. The recombinant virus was stable for 15 passages in cell culture. Pigs vaccinated with CH-1R/pIL-4 produced a similar humoral response to the response elicited by parental virus, but IL-4 level in the supernatant of PBMCs from pigs vaccinated with CH-1R/pIL-4 was significantly higher than the parent virus at 28 days post-immunization (DPI). Flow cytometric (FCM) analysis showed that the percentage of CD4(+)CD8(+) double positive T (DPT) cells in the CH-1R/pIL-4 vaccinated group was significantly higher than the parental virus at 3 and 7 Days Post-Challenge (DPC), and the IL-4 level in the blood significantly increased at 7 DPC. However, the viral load and histopathology did not show significant difference between the two groups. CONCLUSIONS: A recombinant PRRSV expressing porcine IL-4 was rescued and it remained genetically stable in vitro. The recombinant virus induced higher DPT ratios and IL-4 levels in the blood after HP-PRRSV challenge compared to the parental virus in piglets. However, it did not significantly improve protection efficacy of PRRSV vaccine. FAU - Li, Zhijun AU - Li Z AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. zjlzlj@yahoo.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. zjlzlj@yahoo.com. FAU - Wang, Gang AU - Wang G AD - State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agriculture Science, Harbin, 150001, China. wg0381@163.com. FAU - Wang, Yan AU - Wang Y AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. wangyanfushi@163.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. wangyanfushi@163.com. FAU - Zhang, Chong AU - Zhang C AD - State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agriculture Science, Harbin, 150001, China. leofighting@163.com. FAU - Wang, Xinglong AU - Wang X AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. wxlong@nwsuaf.edu.cn. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. wxlong@nwsuaf.edu.cn. FAU - Huang, Baicheng AU - Huang B AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. hbch228@163.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. hbch228@163.com. FAU - Li, Qiongyi AU - Li Q AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. jean_lqy@sina.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. jean_lqy@sina.com. FAU - Li, Liangliang AU - Li L AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. lifeiyang2007@126.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. lifeiyang2007@126.com. FAU - Xue, Biyun AU - Xue B AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. 18717368270@163.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. 18717368270@163.com. FAU - Ding, Peiyang AU - Ding P AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. Dingpeiyang1990@163.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. Dingpeiyang1990@163.com. FAU - Syed, Shahid Faraz AU - Syed SF AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. farazvet@yahoo.com. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. farazvet@yahoo.com. FAU - Wang, Chengbao AU - Wang C AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. wangchengbao@nwsuaf.edu.cn. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. wangchengbao@nwsuaf.edu.cn. FAU - Cai, Xuehui AU - Cai X AD - State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agriculture Science, Harbin, 150001, China. aci139@sina.com. FAU - Zhou, En-Min AU - Zhou EM AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. zhouem@nwsuaf.edu.cn. AD - Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China. zhouem@nwsuaf.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151114 PL - England TA - Virol J JT - Virology journal JID - 101231645 RN - 0 (Adjuvants, Immunologic) RN - 0 (Antibodies, Viral) RN - 0 (Viral Vaccines) RN - 207137-56-2 (Interleukin-4) SB - IM MH - Adjuvants, Immunologic/*biosynthesis/genetics MH - Animals MH - Antibodies, Viral/blood MH - CD4-Positive T-Lymphocytes/immunology MH - CD8-Positive T-Lymphocytes/immunology MH - Cell Line MH - Gene Expression MH - Genomic Instability MH - Histocytochemistry MH - Injections, Intramuscular MH - Interleukin-4/*biosynthesis/genetics MH - Porcine Reproductive and Respiratory Syndrome/immunology/pathology/*prevention & control MH - Porcine respiratory and reproductive syndrome virus/genetics/growth & development/*immunology/*isolation & purification MH - Recombination, Genetic MH - Swine MH - Viral Load MH - Viral Vaccines/administration & dosage/genetics/*immunology/*isolation & purification PMC - PMC4647277 EDAT- 2015/11/18 06:00 MHDA- 2016/06/30 06:00 PMCR- 2015/11/14 CRDT- 2015/11/18 06:00 PHST- 2015/05/12 00:00 [received] PHST- 2015/09/09 00:00 [accepted] PHST- 2015/11/18 06:00 [entrez] PHST- 2015/11/18 06:00 [pubmed] PHST- 2016/06/30 06:00 [medline] PHST- 2015/11/14 00:00 [pmc-release] AID - 10.1186/s12985-015-0380-7 [pii] AID - 380 [pii] AID - 10.1186/s12985-015-0380-7 [doi] PST - epublish SO - Virol J. 2015 Nov 14;12:185. doi: 10.1186/s12985-015-0380-7.