title: Comparative analysis of cytokine transcript profiles within mediastinal lymph node compartments of pigs after infection with porcine reproductive and respiratory syndrome genotype 1 strains differing in pathogenicity.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) induces a weak immune response enabling it to persist in different organs of infected pigs. This has been attributed to the ability of PRRSV to influence the induction of cytokine responses. In this study, we investigated the cytokine transcriptional profiles in different compartments of the mediastinal lymph node of pigs infected with three genotype 1 PRRSV strains of differing pathogenicity: the low virulence prototype Lelystad virus (LV), and UK field strain 215-06 and the highly virulent subtype 3 SU1-Bel isolate from Belarus. We have used a combination of laser capture micro-dissection (LCM) followed by real time quantitative PCR (RT-qPCR) and immunohistochemical (IHC) detection of immune cell markers (CD3, CD79a and MAC387) and RT-qPCR quantification of PRRSV and cytokine transcripts. Compared to mock infected pigs, we found a significant downregulation of TNF-α and IFN-α in follicular and interfollicular areas of the mediastinal lymph node from 3 days post-infection (dpi) in animals infected with all three strains. This was accompanied by a transient B cell depletion and T cell and macrophage infiltration in the follicles together with T cell depletion in the interfollicular areas. A delayed upregulation of IFN-γ and IL-23p19 was observed mainly in the follicles. The PRRSV load was higher in all areas and time-points studied in the animals infected with the SU1-Bel strain. This paper describes the first application of LCM to study the cytokine transcript profiles and virus distribution in different compartments of the lymph node of pigs.
authors.affiliation: Department of Anatomy and Comparative Pathology, Faculty of Veterinary Medicine, Murcia University, "Mare Nostrum Excellence Campus - 3738", 30100, Murcia, Spain. ogn14346@um.es.
authors.forename: Obdulio
authors.initials: O
authors.lastname: García-Nicolás
authors.affiliation: Department of Bacteriology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. ruben.rosales@apha.gsi.gov.uk.
authors.forename: Rubén S
authors.initials: RS
authors.lastname: Rosales
authors.affiliation: Department of Anatomy and Comparative Pathology, Faculty of Veterinary Medicine, Murcia University, "Mare Nostrum Excellence Campus - 3738", 30100, Murcia, Spain. pallares@um.es.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Pallarés
authors.affiliation: Red de Grupos de Investigación Recursos Faunísticos, Facultad de Veterinaria, Universidad de Extremadura, 10003, Cáceres, Spain. driscope@gmail.com.
authors.forename: David
authors.initials: D
authors.lastname: Risco
authors.affiliation: Spanish National Center of Biotechnology (CSIC), C/Darwin 3, Campus de Cantoblanco, 28049, Madrid, Spain. jjquere@um.es.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Quereda
authors.affiliation: Department of Virology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. Simon.Graham@apha.gsi.gov.uk.
authors.forename: Simon P
authors.initials: SP
authors.lastname: Graham
authors.affiliation: Department of Virology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. Jean-pierre.frossard@apha.gsi.gov.uk.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Frossard
authors.affiliation: Department of Virology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. Sophie.morgan@yahoo.co.uk.
authors.forename: Sophie B
authors.initials: SB
authors.lastname: Morgan
authors.affiliation: Department of Virology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. falko.steinbach@apha.gsi.gov.uk.|Department of Pathology and Infectious Diseases, School of Veterinary Medicine, University of Surrey, Guildford, GU2 7TE, UK. falko.steinbach@apha.gsi.gov.uk.
authors.forename: Falko
authors.initials: F
authors.lastname: Steinbach
authors.affiliation: Department of Virology, Animal and Plant Health Agency, Addlestone, Surrey, KT15 3NB, UK. Trevor.drew@apha.gsi.gov.uk.
authors.forename: Trevor W
authors.initials: TW
authors.lastname: Drew
authors.affiliation: Department of Pathology, Animal and Plant Health Agency, Addlestone, KT15 3NB, UK. Tony.strickland@apha.gsi.gov.uk.
authors.forename: Tony S
authors.initials: TS
authors.lastname: Strickland
authors.affiliation: Department of Pathology, Animal and Plant Health Agency, Addlestone, KT15 3NB, UK. f.salguerobodes@surrey.ac.uk.|Department of Pathology and Infectious Diseases, School of Veterinary Medicine, University of Surrey, Guildford, GU2 7TE, UK. f.salguerobodes@surrey.ac.uk.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Salguero
mesh_terms: D000818:Animals; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D008198:Lymph Nodes / Q000821:virology*; D008482:Mediastinum / Q000821:virology; D019318:Porcine Reproductive and Respiratory Syndrome / Q000235:genetics / Q000276:immunology* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000472:pathogenicity*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013552:Swine; D059467:Transcriptome; D014774:Virulence
chemical_list: D016207:Cytokines; D012367:RNA, Viral