Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG
The G-H loop of the foot-and-mouth disease virus (FMDV) virion contains certain dominant immunogenic epitopes, as well as an arginine-glycine-aspartic acid (RGD) motif that is recognized by cell surface integrin receptors. Previous experiments indicate that it is critical to maintain virus structura...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2018-07-01
|
Series: | Journal of Integrative Agriculture |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311918619161 |
id |
doaj-e637c8854cdf4e988f3c58b9906ce9a2 |
---|---|
record_format |
Article |
spelling |
doaj-e637c8854cdf4e988f3c58b9906ce9a22021-06-08T04:39:51ZengElsevierJournal of Integrative Agriculture2095-31192018-07-0117716551666Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAGYuan-yuan ZHU0Xing-qi ZOU1Hui-fang BAO2Pu SUN3Xue-qing MA4Zai-xin LIU5Hong-jie FAN6Qi-zu ZHAO7MOE Joint International Research Laboratory of Animal Health and Food Safety/College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R. China; China Institute of Veterinary Drug Control, Beijing 100081, P.R. ChinaChina Institute of Veterinary Drug Control, Beijing 100081, P.R. ChinaLanzhou Veterinary Research Institute, China Academy of Agricultrual Sciences, Lanzhou 730046, P.R. ChinaLanzhou Veterinary Research Institute, China Academy of Agricultrual Sciences, Lanzhou 730046, P.R. ChinaLanzhou Veterinary Research Institute, China Academy of Agricultrual Sciences, Lanzhou 730046, P.R. ChinaLanzhou Veterinary Research Institute, China Academy of Agricultrual Sciences, Lanzhou 730046, P.R. ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety/College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R. China; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, P.R. China; Correspondence FAN Hong-jie, Tel/Fax: +86-25-84396219China Institute of Veterinary Drug Control, Beijing 100081, P.R. China; ZHAO Qi-zu, Tel/Fax: +86-10-62103670The G-H loop of the foot-and-mouth disease virus (FMDV) virion contains certain dominant immunogenic epitopes, as well as an arginine-glycine-aspartic acid (RGD) motif that is recognized by cell surface integrin receptors. Previous experiments indicate that it is critical to maintain virus structural integrity when inserting an exogenous epitope into the surface of an FMDV structural protein. However, it remains to be determined how factors such as different insertion positions affect interactions among the virus, cells and host immune system. In this study, one infectious cDNA clone of the swine FMDV Cathay topotype strain O/CHA/90 was constructed. Then, a FLAG marker (DYKDDDDK) was inserted upstream (–4) or downstream (+10) of the RGD motif to generate tagged viruses vFLAG-O/CHA/90 or vO/CHA/90-FLAG, investigating the possibility of expressing foreign antigen and effect on its immunogenicity. Compared to the parental virus, both tagged viruses exhibited similar plaque phenotypes, suckling mouse pathogenicity and antigenicity. Additionally, the FLAG tag insertion position did not change the use of integrin-mediated cell entry by the tagged viruses. Interestingly, both tagged vaccines protected pigs against challenge with the parental virus O/CHA/90 and induced immune responses against FMDV in BALB/c mice and pigs, but only vaccination with vFLAG-O/CHA/90 generated anti-FLAG antibodies. Our findings demonstrated that two sites (RGD–4 and RGD+10) tolerated the insertion of an exogenous gene in the swine FMDV O/CHA/90 strain. However, only RGD–4 was a novel and appropriate inserting site which could tolerate exogenous FLAG. The resultant tagged virus is a promising candidate for FMD vaccine which can be differentiating infected from vaccinated animals (DIVA).http://www.sciencedirect.com/science/article/pii/S2095311918619161FMDVG-H loopinsertion sitesFLAG-tagged virusesreverse genetics |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuan-yuan ZHU Xing-qi ZOU Hui-fang BAO Pu SUN Xue-qing MA Zai-xin LIU Hong-jie FAN Qi-zu ZHAO |
spellingShingle |
Yuan-yuan ZHU Xing-qi ZOU Hui-fang BAO Pu SUN Xue-qing MA Zai-xin LIU Hong-jie FAN Qi-zu ZHAO Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG Journal of Integrative Agriculture FMDV G-H loop insertion sites FLAG-tagged viruses reverse genetics |
author_facet |
Yuan-yuan ZHU Xing-qi ZOU Hui-fang BAO Pu SUN Xue-qing MA Zai-xin LIU Hong-jie FAN Qi-zu ZHAO |
author_sort |
Yuan-yuan ZHU |
title |
Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG |
title_short |
Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG |
title_full |
Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG |
title_fullStr |
Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG |
title_full_unstemmed |
Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG |
title_sort |
insertion site of flag on foot-and-mouth disease virus vp1 g-h loop affects immunogenicity of flag |
publisher |
Elsevier |
series |
Journal of Integrative Agriculture |
issn |
2095-3119 |
publishDate |
2018-07-01 |
description |
The G-H loop of the foot-and-mouth disease virus (FMDV) virion contains certain dominant immunogenic epitopes, as well as an arginine-glycine-aspartic acid (RGD) motif that is recognized by cell surface integrin receptors. Previous experiments indicate that it is critical to maintain virus structural integrity when inserting an exogenous epitope into the surface of an FMDV structural protein. However, it remains to be determined how factors such as different insertion positions affect interactions among the virus, cells and host immune system. In this study, one infectious cDNA clone of the swine FMDV Cathay topotype strain O/CHA/90 was constructed. Then, a FLAG marker (DYKDDDDK) was inserted upstream (–4) or downstream (+10) of the RGD motif to generate tagged viruses vFLAG-O/CHA/90 or vO/CHA/90-FLAG, investigating the possibility of expressing foreign antigen and effect on its immunogenicity. Compared to the parental virus, both tagged viruses exhibited similar plaque phenotypes, suckling mouse pathogenicity and antigenicity. Additionally, the FLAG tag insertion position did not change the use of integrin-mediated cell entry by the tagged viruses. Interestingly, both tagged vaccines protected pigs against challenge with the parental virus O/CHA/90 and induced immune responses against FMDV in BALB/c mice and pigs, but only vaccination with vFLAG-O/CHA/90 generated anti-FLAG antibodies. Our findings demonstrated that two sites (RGD–4 and RGD+10) tolerated the insertion of an exogenous gene in the swine FMDV O/CHA/90 strain. However, only RGD–4 was a novel and appropriate inserting site which could tolerate exogenous FLAG. The resultant tagged virus is a promising candidate for FMD vaccine which can be differentiating infected from vaccinated animals (DIVA). |
topic |
FMDV G-H loop insertion sites FLAG-tagged viruses reverse genetics |
url |
http://www.sciencedirect.com/science/article/pii/S2095311918619161 |
work_keys_str_mv |
AT yuanyuanzhu insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT xingqizou insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT huifangbao insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT pusun insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT xueqingma insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT zaixinliu insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT hongjiefan insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag AT qizuzhao insertionsiteofflagonfootandmouthdiseasevirusvp1ghloopaffectsimmunogenicityofflag |
_version_ |
1721390540318048256 |