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Interleukin-4, interleukin-5, and interleukin-13 gene expression in cultured mononuclear cells from porcine circovirus type 2–vaccinated pigs after cells were challenged with porcine circovirus type 2 open reading frame 2 antigen

Juan J. QueredaDepartment of Animal Production, Faculty of Veterinary Medicine, University of Murcia, 30100 Murcia, Spain.

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Guillermo RamisDepartment of Animal Production, Faculty of Veterinary Medicine, University of Murcia, 30100 Murcia, Spain.

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Francisco J. PallarésDepartment of Anatomical Pathology, Faculty of Veterinary Medicine, University of Murcia, 30100 Murcia, Spain.

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Ludivine ChapatMerial SAS, 29 Ave Tony Garnier, BP7123, 69348 Lyon, France.

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Anne GoubierGenticel, Prologue-Biotech, 515 Rue Pierre et Marie Curie BP 28262, 31682 Labège-Innopole Cedex, France.

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François JoiselMerial SAS, 29 Ave Tony Garnier, BP7123, 69348 Lyon, France.

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Catherine CharreyreMerial SAS, 29 Ave Tony Garnier, BP7123, 69348 Lyon, France.

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David VillarFaculty of Agricultural Sciences, University of Antioquia, 75 Medellin, Colombia.

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Antonio MuõozDepartment of Animal Production, Faculty of Veterinary Medicine, University of Murcia, 30100 Murcia, Spain.

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Abstract

Objective—To characterize the kinetics of interleukin (IL)-4, IL-5, and IL-13 secretion in peripheral blood and lymph node mononuclear cells isolated from porcine circovirus type 2 (PCV2)–vaccinated pigs after cells were challenged with PCV2 open reading frame 2 antigen.

Animals—10 pigs.

Procedures—5 pigs were vaccinated with a PCV2 vaccine and received a booster dose 3 weeks later. They were kept together with a similar group of 5 nonvaccinated pigs that served as controls. One week after the second vaccination, peripheral blood mononuclear cells (PBMCs) and excised retropharyngeal lymph node mononuclear cells (LNMCs) were isolated and cultured. Cells were then challenged by exposure to PCV2 open reading frame 2 and evaluated at 2, 12, 24, and 48 hours to determine the expression of IL-4, IL-5, and IL-13 via quantitative PCR assay. Changes in gene expression were analyzed relative to the results from analysis of the sample at 0 hours (calibrator).

Results—All ILs were upregulated differently in LNMCs and PBMCs from vaccinated pigs. Lymph node mononuclear cells from vaccinated animals produced significantly more IL-4 mRNA than did PBMCs at 2, 12, and 48 hours (relative change: 2.8 vs −3.6, 13.0 vs 3.6, and 9.8 vs 1.8, respectively) and more IL-5 mRNA at 2, 12, 24, and 48 hours (relative change: 1. 2 vs −4.8, 2.2 vs 0.2, 3.2 vs −1.9, and 4.0 vs −3.6, respectively). Interleukin-13 mRNA reached its highest concentration at 24 hours but was 11.9-fold higher in PBMCs than in LNMCs.

Conclusions and Clinical Relevance—Results supported the importance of IL-4, IL-5, and IL-13 in pigs, suggesting that PBMCs and LNMCs express cytokines in a tissue-specific manner.

Abstract

Objective—To characterize the kinetics of interleukin (IL)-4, IL-5, and IL-13 secretion in peripheral blood and lymph node mononuclear cells isolated from porcine circovirus type 2 (PCV2)–vaccinated pigs after cells were challenged with PCV2 open reading frame 2 antigen.

Animals—10 pigs.

Procedures—5 pigs were vaccinated with a PCV2 vaccine and received a booster dose 3 weeks later. They were kept together with a similar group of 5 nonvaccinated pigs that served as controls. One week after the second vaccination, peripheral blood mononuclear cells (PBMCs) and excised retropharyngeal lymph node mononuclear cells (LNMCs) were isolated and cultured. Cells were then challenged by exposure to PCV2 open reading frame 2 and evaluated at 2, 12, 24, and 48 hours to determine the expression of IL-4, IL-5, and IL-13 via quantitative PCR assay. Changes in gene expression were analyzed relative to the results from analysis of the sample at 0 hours (calibrator).

Results—All ILs were upregulated differently in LNMCs and PBMCs from vaccinated pigs. Lymph node mononuclear cells from vaccinated animals produced significantly more IL-4 mRNA than did PBMCs at 2, 12, and 48 hours (relative change: 2.8 vs −3.6, 13.0 vs 3.6, and 9.8 vs 1.8, respectively) and more IL-5 mRNA at 2, 12, 24, and 48 hours (relative change: 1. 2 vs −4.8, 2.2 vs 0.2, 3.2 vs −1.9, and 4.0 vs −3.6, respectively). Interleukin-13 mRNA reached its highest concentration at 24 hours but was 11.9-fold higher in PBMCs than in LNMCs.

Conclusions and Clinical Relevance—Results supported the importance of IL-4, IL-5, and IL-13 in pigs, suggesting that PBMCs and LNMCs express cytokines in a tissue-specific manner.

Contributor Notes

Dr. Quereda's present address is Spanish National Center of Biotechnology—CSIC, Darwin 3, Campus de Cantoblanco, 28049 Madrid, Spain.

Dr. Quereda was supported by a doctoral grant from the Spanish Ministry of Education and Science (AP-2005-3468).

Ludivine Chapat, François Joisel, and Catherine Charreyre are employees of Merial SAS.

Address correspondence to Dr. Quereda (jjquere@um.es).