Local innate responses and protective immunity after intradermal immunization with bovine viral diarrhea virus E2 protein formulated with a combination adjuvant in cattle

doi:10.1016/j.vaccine.2017.05.029

Vaccine

Volume 35, Issue 27, 14 June 2017, Pages 3466-3473

https://doi.org/10.1016/j.vaccine.2017.05.029 Get rights and content

Abstract

Bovine viral diarrhea virus (BVDV) is one of the most serious pathogens in cattle. Recently, we developed a novel adjuvant platform (TriAdj) that includes a toll-like receptor 3 agonist, poly (I:C); an innate defense regulatory peptide; and water-soluble polymer, poly[di(sodiumcarboxylatoethylphenoxy)]-phosphazene (PCEP). To develop a needle-free intradermal (ID) subunit vaccine, the BVDV type-2 E2 protein was formulated with TriAdj, and immune protection was evaluated in calves against a BVDV-2 strain. Intradermal delivery of E2/TriAdj elicited robust virus neutralizing antibodies and cell-mediated immune responses including CD4⁺ and CD8⁺ T-cell responses. The development of CD8⁺ T-cell responses in vaccinated calves indicates that TriAdj promotes cross-presentation. Upon challenge with virulent BVDV-2, the vaccinated calves showed no weight loss, leukopenia or virus shedding, and almost no temperature increase, in contrast to the control animals, which had severe clinical disease and shed virus for three to six days in nasal fluids and white blood cells. Intradermal vaccination was shown to attract various immune cell populations including dendritic cells, the most important antigen presenting cells. These data demonstrate that ID delivery is suitable as an administration route in cattle and that ID delivered, TriAdj-formulated E2 can protect cattle from BVDV-2.

Introduction

Bovine viral diarrhea virus (BVDV) is a Pestivirus within the Flaviviridae family, and consists of two subtypes; a third subtype, BVDV-3, has been proposed [1]. Acute disease in BVDV-infected cattle is initially acknowledged based on fever, increased respiratory rate, diarrhea, and low white blood cell (WBC) counts [2]. BVDV-2 strains sometimes cause more acute infections than BVDV-1 with higher fever and severe diarrhea, as well as lower WBCs and platelets; this may result in death [3]. BVDV also is a major contributor to the bovine respiratory disease complex [4]. BVDV infections in pregnant cows may result in persistently infected calves, which may develop mucosal disease [3]. In the past few decades, adversely affected health and productivity of cattle have established BVDV as a virulent pathogen of high economic importance for the dairy and cattle industries [4].

BVDV is classified as a positive-sense single-stranded RNA virus. The virions contain a nucleocapsid surrounded by an envelope with glycoproteins E^RNS, E1 and E2 [5], [6]. Among these glycoproteins, E2 is responsible for entry of the virus into the host cells [7], and is the major protective antigen, capable of inducing virus neutralizing (VN) antibodies and protection from BVDV challenge in cattle [8].

Since strong adjuvants are needed for optimal efficacy of subunit vaccines, we formulated the E2 protein with a combination adjuvant (TriAdj) consisting of a toll-like receptor-3 (TLR3) agonist, poly(I:C); an innate defense regulatory peptide, IDR-1002; and water-soluble polymer, poly[di(sodiumcarboxylatoethylphenoxy)]-phosphazene (PCEP). Poly(I:C) induces Th1-biased immune responses and promotes cross-presentation [9]. Immunomodulation by a synthetic IDR peptide can strengthen the host defense against microbial infections by recruiting activated immune cells [10], [11]. This cationic antimicrobial peptide also regulates innate immune responses at the site of infection or immunization by enhancing monocyte infiltration [12]. PCEP has immunostimulatory properties and induces balanced immune responses after co-administration with antigen [13].

Previously, E2 protein formulated with TriAdj (E2/TriAdj) was found to be protective in cattle when delivered intramuscularly (IM) [2]. However, intradermal (ID) vaccine delivery is of interest as epidermal and dermal layers of skin are rich in antigen-presenting cells (APCs) [14]. Therefore, vaccine delivery in these layers is expected to be as efficient, or even more, compared to delivery in muscle or subcutaneous tissue [15]. Furthermore, ID delivery avoids tissue damage to the muscle, which may lead to reduced meat quality and economic losses. In view of these advantages, the goal of this study was to develop an effective ID subunit BVDV-2 vaccine.

Section snippets

Vaccine, immunizations and challenge

A truncated, secreted version of the BVDV-2 E2 protein (strain Q140; accession number AAB01811; amino acids 41-383) was produced in a transient HEK293 cell expression system, and purified and characterized as described previously [2]. BVDV-seronegative seven to eight month-old Angus-Hereford cross-bred calves were randomly allocated to three treatment groups, designated IM, ID, and Placebo, with five animals per group. The calves were immunized IM or ID with a needle-free device, a Biojector®,

Humoral and cell-mediated immune responses elicited by E2 formulated with TriAdj

To determine the impact of delivery route on vaccine efficacy, calves were immunized with E2/TriAdj, either IM or ID, followed by challenge with BVDV-2. After vaccination a significant increase in IgG titers was observed in both the IM and ID groups when compared to the Placebo group (Fig. 1A). The titers were further enhanced after BVDV-2 challenge. However, there was no difference between the IgG titers of the IM and ID groups. The VN antibody titers specific for strain 1373 were also

Discussion

In this study, ID delivery of E2/TriAdj vaccine induced robust humoral and cell-mediated immune responses, as well as protection from BVDV-2 in cattle, equivalent to those induced by IM delivery. Moreover, infiltration of relevant immune cell populations including APCs were observed after ID delivery. This is the first report showing efficacy of ID delivery of an adjuvanted E2 protein subunit vaccine in a vaccination-challenge trial in cattle. These data support this route as very suitable and

Acknowledgements

We thank Elisa Martinez and Wayne Connor for assistance with the qRT-PCR, Sarah Mackenzie-Dyck for help with the cell isolations and Glenn Hamonic for training in cryo-sectioning the bovine skin tissues. We also thank Don Wilson, Brock Evans, and Sherry Tetland for care and handling of the calves. Financial support was provided by Saskatchewan Agricultural Development Fund (Grant number 20140232) and Alberta Livestock and Meat Agency (Grant number 2015B011R), Canada. This is manuscript 794 of

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Local innate responses and protective immunity after intradermal immunization with bovine viral diarrhea virus E2 protein formulated with a combination adjuvant in cattle

Abstract

Introduction

Section snippets

Vaccine, immunizations and challenge

Humoral and cell-mediated immune responses elicited by E2 formulated with TriAdj

Discussion

Acknowledgements

Virology

Vaccine.

Vet Clin North Am Food Anim Pract.

Veter Clin North Am Food Anim Pract.

Vaccine.

Biochem Biophys Res Commun.

Vaccine.

Vaccine.

Vaccine.

Veter Microbiol.

Veter Clin North Am Food Anim Pract.

Res Veter Sci.

Vaccine.

Vaccine.

Vaccine.

Eur J Pharm Biopharm: Off J Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik eV.

Veterinary medicine—a textbook of the diseases of cattle, horses, sheep, pigs and goats, 10th edition

Can Veter J.