title: Characterization of a Broadly Reactive Anti-CD40 Agonistic Monoclonal Antibody for Potential Use as an Adjuvant.
abstract: Lack of safe and effective adjuvants is a major hindrance to the development of efficacious vaccines. Signaling via CD40 pathway leads to enhanced antigen processing and presentation, nitric oxide expression, pro-inflammatory cytokine expression by antigen presenting cells, and stimulation of B-cells to undergo somatic hypermutation, immunoglobulin class switching, and proliferation. Agonistic anti-CD40 antibodies have shown promising adjuvant qualities in human and mouse vaccine studies. An anti-CD40 monoclonal antibody (mAb), designated 2E4E4, was identified and shown to have strong agonistic effects on primary cells from multiple livestock species. The mAb recognize swine, bovine, caprine, and ovine CD40, and evoked 25-fold or greater proliferation of peripheral blood mononuclear cells (PBMCs) from these species relative to cells incubated with an isotype control (p<0.001). In addition, the mAb induced significant nitric oxide (p<0.0001) release by bovine macrophages. Furthermore, the mAb upregulated the expression of MHC-II by PBMCs, and stimulated significant (p<0.0001) IL-1α, IL6, IL-8, and TNF-α expression by PBMCs. These results suggest that the mAb 2E4E4 can target and stimulate cells from multiple livestock species and thus, it is a potential candidate for adjuvant development. This is the first study to report an anti-swine CD40 agonistic mAb that is also broadly reactive against multiple species.
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Cameron
authors.identifier: http://orcid.org/0000-0003-0824-3106
authors.initials: C
authors.lastname: Martin
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Suryakant D
authors.initials: SD
authors.lastname: Waghela
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Shehnaz
authors.initials: S
authors.lastname: Lokhandwala
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Andy
authors.initials: A
authors.lastname: Ambrus
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Jocelyn
authors.initials: J
authors.lastname: Bray
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Christina
authors.initials: C
authors.lastname: Vuong
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Vanitha
authors.initials: V
authors.lastname: Vinodkumar
authors.affiliation: Zoetis, Kalamazoo, Michigan, United States of America.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Dominowski
authors.affiliation: Zoetis, Kalamazoo, Michigan, United States of America.
authors.forename: Sharath
authors.initials: S
authors.lastname: Rai
authors.affiliation: Zoetis, Kalamazoo, Michigan, United States of America.
authors.forename: Duncan
authors.initials: D
authors.lastname: Mwangi
authors.affiliation: Zoetis, Kalamazoo, Michigan, United States of America.
authors.forename: Dennis L
authors.initials: DL
authors.lastname: Foss
authors.affiliation: Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Waithaka
authors.initials: W
authors.lastname: Mwangi
mesh_terms: D000276:Adjuvants, Immunologic / Q000627:therapeutic use*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology*; D002417:Cattle / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D005802:Genes, MHC Class II / Q000276:immunology; D006041:Goats / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012756:Sheep / Q000276:immunology; D013552:Swine / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D016207:Cytokines