title: Generation of a soluble recombinant trimeric form of bovine CD40L and its potential use as a vaccine adjuvant in cows.
abstract: Vaccination is the most cost-effective way to control infectious diseases in cattle. However, many infectious diseases leading to severe economical losses worldwide still remain for which a really effective and safe vaccine is not available. These diseases are most often due to intracellular pathogens such as bacteria or viruses, which are, by their localization, protected from antibiotics and/or CD4(+) T cell-dependent humoral responses. We therefore postulated that strategies leading to induction of not only CD4(+) T cell responses but also CD8(+) cytotoxic T lymphocyte (CTL) responses against infected cells should be privileged in the development of new vaccines against problematic intracellular pathogens in bovines. CD40 signaling in antigen-presenting cells may lead to the induction of robust CD4-independent CTL responses and several studies, especially in mice, have used CD40 stimulation to promote CD8(+) T cell-mediated immunity. For example, we have recently shown that immunization of mice with heat-killed Staphylococcus aureus (HKSA) and agonistic anti-CD40 monoclonal antibodies elicits strong CTL responses capable of protecting mice from subsequent staphylococcal mastitis. Unfortunately, there is at present no tool available to efficiently stimulate CD40 in cattle. In this study, we therefore first produced a soluble recombinant trimeric form of the natural bovine CD40 ligand (sboCD40LT). We then observed that sboCD40LT was able to potently stimulate bovine cells in vitro. Finally, we provide evidence that immunization of cows with sboCD40LT combined with HKSA was able to significantly increase the number of both HKSA-specific CD4(+) and CD8(+) T cells in the draining lymph nodes. In conclusion, we suggest that this new molecular tool could help in the development of vaccine strategies against bovine diseases caused by intracellular pathogens.
authors.affiliation: Laboratory of Cellular and Molecular Immunology, GIGA-Research, University of Liège, 1 Avenue de l'Hôpital (B34), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Julien
authors.initials: J
authors.lastname: Pujol
authors.affiliation: Laboratory of Biological Macromolecules, Centre for Protein Engineering, Faculty of Sciences, University of Liège, 3 Allée de la Chimie (B6a), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Bouillenne
authors.affiliation: Department of Animal Production, Faculty of Veterinary Medicine, University of Liège, 20 Boulevard de Colonster (B43), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Farnir
authors.affiliation: Department of Animal Production, Faculty of Veterinary Medicine, University of Liège, 20 Boulevard de Colonster (B43), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Dufrasne
authors.affiliation: Department of Infectious and Parasitic Diseases-Bacteriology, Faculty of Veterinary Medicine, University of Liège, 20 Boulevard de Colonster (B43), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Jacques
authors.initials: J
authors.lastname: Mainil
authors.affiliation: Laboratory of Biological Macromolecules, Centre for Protein Engineering, Faculty of Sciences, University of Liège, 3 Allée de la Chimie (B6a), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Moreno
authors.initials: M
authors.lastname: Galleni
authors.affiliation: Laboratory of Cellular and Molecular Immunology, GIGA-Research, University of Liège, 1 Avenue de l'Hôpital (B34), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Pierre
authors.initials: P
authors.lastname: Lekeux
authors.affiliation: Laboratory of Cellular and Molecular Immunology, GIGA-Research, University of Liège, 1 Avenue de l'Hôpital (B34), Sart Tilman, Liège, B-4000, Belgium.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Bureau
authors.affiliation: Laboratory of Cellular and Molecular Immunology, GIGA-Research, University of Liège, 1 Avenue de l'Hôpital (B34), Sart Tilman, Liège, B-4000, Belgium. Electronic address: laurence.fievez@ulg.ac.be.
authors.forename: Laurence
authors.initials: L
authors.lastname: Fiévez
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D019013:CD40 Antigens / Q000378:metabolism; D023201:CD40 Ligand / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology / Q000517:prevention & control*; D003001:Cloning, Molecular; D042783:Endothelial Cells / Q000276:immunology; D005260:Female; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D051379:Mice; D020836:Protein Structure, Quaternary; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D013211:Staphylococcus aureus / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014611:Vaccination / Q000662:veterinary*
chemical_list: D000276:Adjuvants, Immunologic; D019013:CD40 Antigens; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand
keywords: Bovine CD40L; CD40; CFSE; GST; HKSA; HSV; ME; PLN; Recombinant protein; Vaccination; carboxyfluorescein succinimidyl ester; glutathione S-transferase; heat-killed Staphylococcus aureus; herpes simplex virus; precrural lymph node; sboCD40LT; soluble recombinant trimeric form of bovine CD40L; β-mercaptoethanol