New Formulation of a Subunit Vaccine Candidate against Lawsonia intracellularis Increases Humoral and Cellular Immune Responses

Author:

Salazar Santiago1ORCID,Starck María Francisca1,Villegas Milton F.1,Acosta Jannel1,Sánchez Oliberto2,Ramos Eduardo1,Nova-Lamperti Estefanía3ORCID,Toledo Jorge R.1,Gädicke Paula4ORCID,Ruiz Álvaro4ORCID,González Alaín5ORCID,Montesino Raquel1

Affiliation:

1. Biotechnology and Biopharmaceuticals Laboratory, Pathophysiology Department, School of Biological Sciences, Universidad de Concepción, Victor Lamas 1290, Concepción P.O. Box 160-C, Chile

2. Pharmacology Department, School of Biological Sciences, Universidad de Concepción, Victor Lamas 1290, Concepción P.O. Box 160-C, Chile

3. Molecular and Translational Immunology Laboratory, Clinical Biochemistry and Immunology Department, Pharmacy Faculty, Universidad de Concepción, Victor Lamas 1290, Concepción P.O. Box 160-C, Chile

4. Pathology and Preventive Medicine Department, School of Veterinary Sciences, Universidad de Concepción, Avenida Vicente Méndez 595, Chillan P.O. Box 537, Chile

5. Faculty of Basic Sciences, University of Medellin, Cra. 87 No. 30-65, Medellin P.C. 050026, Antioquia, Colombia

Abstract

Previously, we designed a subunit vaccine candidate based on three L. intracellularis antigens with promising results in pigs. In this study, antigens were produced individually to achieve an even antigen ratio in the formulation. The emulsion characterization included the drop size and the mechanical and thermal stability. Immune response was evaluated by indirect and sandwich ELISAs, qPCR, and flow cytometry. The vaccine candidate’s safety was assessed by histopathology and monitoring the clinical behavior of animals. The average production yielded for the chimeric antigen as inclusion bodies was around 75 mg/L. The formulation showed mechanical and thermal stability, with a ratio Hu/Ho > 0.85 and a drop size under 0.15 nm. Antigens formulated at a ratio of 1:1:1 induced a significant immune response in inoculated pigs that persisted until the end of the experiment (week 14). The dose of 200 μg significantly activated cellular response measured by transcriptional and translational levels of cytokines. The cell proliferation assay revealed an increment of lymphocytes T CD4+ at the same dose. Animals gained weight constantly and showed proper clinical behavior during immunization assays. This research demonstrated the immunological robustness of the new subunit vaccine candidate against Porcine Proliferative Enteropathy evenly formulated with three chimeric antigens of L. intracellularis.

Funder

Scientific and Technological Development Support Fund

Publisher

MDPI AG

Subject

Pharmacology (medical),Infectious Diseases,Drug Discovery,Pharmacology,Immunology

Reference46 articles.

1. Defining the full costs of endemic porcine proliferative enteropathy;McOrist;Vet. J.,2005

2. Recent advances in understanding the pathogenesis of Lawsonia intracellularis infections;Vannucci;Vet. Pathol.,2014

3. Early pathogenesis in porcine proliferative enteropathy caused by Lawsonia intracellularis;Boutrup;J. Comp. Pathol.,2010

4. Proliferative enteropathy;Lawson;J. Comp. Pathol.,2000

5. Zimmerman, J.J., Karriker, L.A., Ramirez, A., Schwartz, K.J., and Stevenson, G.W. (2012). Diseases of Swine, John Wiley & Sons, Inc.. [10th ed.].

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