Research Article: Priming Cross-Protective Bovine Viral Diarrhea Virus-Specific Immunity Using Live-Vectored Mosaic Antigens

Date Published: January 18, 2017

Publisher: Public Library of Science

Author(s): Shehnaz Lokhandwala, Xin Fang, Suryakant D. Waghela, Jocelyn Bray, Leo M. Njongmeta, Andy Herring, Karim W. Abdelsalam, Christopher Chase, Waithaka Mwangi, Paulo Lee Ho.


Bovine viral diarrhea virus (BVDV) plays a key role in bovine respiratory disease complex, which can lead to pneumonia, diarrhea and death of calves. Current vaccines are not very effective due, in part, to immunosuppressive traits and failure to induce broad protection. There are diverse BVDV strains and thus, current vaccines contain representative genotype 1 and 2 viruses (BVDV-1 & 2) to broaden coverage. BVDV modified live virus (MLV) vaccines are superior to killed virus vaccines, but they are susceptible to neutralization and complement-mediated destruction triggered by passively acquired antibodies, thus limiting their efficacy. We generated three novel mosaic polypeptide chimeras, designated NproE2123; NS231; and NS232, which incorporate protective determinants that are highly conserved among BVDV-1a, 1b, and BVDV-2 genotypes. In addition, strain-specific protective antigens from disparate BVDV strains were included to broaden coverage. We confirmed that adenovirus constructs expressing these antigens were strongly recognized by monoclonal antibodies, polyclonal sera, and IFN-γ-secreting T cells generated against diverse BVDV strains. In a proof-of-concept efficacy study, the multi-antigen proto-type vaccine induced higher, but not significantly different, IFN-γ spot forming cells and T-cell proliferation compared to a commercial MLV vaccine. In regards to the humoral response, the prototype vaccine induced higher BVDV-1 specific neutralizing antibody titers, whereas the MLV vaccine induced higher BVDV-2 specific neutralizing antibody titers. Following BVDV type 2a (1373) challenge, calves immunized with the proto-type or the MLV vaccine had lower clinical scores compared to naïve controls. These results support the hypothesis that a broadly protective subunit vaccine can be generated using mosaic polypeptides that incorporate rationally selected and validated protective determinants from diverse BVDV strains. Furthermore, regarding biosafety of using a live vector in cattle, we showed that recombinant human adenovirus-5 was cleared within one week following intradermal inoculation.

Partial Text

Bovine viral diarrhea virus (BVDV), an infectious pathogen that is prevalent in cattle herds globally, is a key agent responsible for causing Bovine Respiratory Disease Complex (BRDC) [1]. Infection with BVDV can cause severe diarrhea, respiratory disease, immunosuppression, abortion, congenital malformations, and birth of persistently infected (PI) calves, which play a major role in virus transmission in herds [2]. Immunosuppression caused by acute infection of unprotected calves allows secondary infections to establish and cause pneumonia or enteritis [3]. The secondary infections are responsible for high rates of morbidity and mortality, and it is estimated that the U.S. livestock industry loses >$1billion annually due to BRDC [4, 5].

The purpose of this study was to develop an efficacious prototype BVDV vaccine which a) overcomes the several disadvantages associated with the MLV vaccine mentioned previously and b) provides broad protection against multiple BVDV genotypes. To this end, we designed mosaic polypeptide consensus sequences of highly immunogenic BVDV antigens such as Npro, E2 glycoprotein and the Nonstructural protein 2–3 based on multiple genotypes. We selected live replication deficient adenovirus as a vector for delivery of these antigens to prime strong humoral as well as cell mediated immune responses. Polyclonal anti-BVDV sera and monoclonal anti-E2 antibodies strongly recognized these mosaic antigens by immunocytometric analysis. Furthermore, PBMCs from BVDV immune steers proliferated strongly upon stimulation by these mosaic antigens. The above outcomes confirmed the authenticity of both B-cell and T-cell epitopes in all the mosaic antigens.




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