35 research outputs found
Immunization of pigs with replication-incompetent adenovirus-vectored African swine fever virus multi-antigens induced humoral immune responses but no protection following contact challenge
IntroductionAfrican swine fever virus (ASFV) is a pathogen of great economic importance given that continues to threaten the pork industry worldwide, but there is no safe vaccine or treatment available. Development of a vaccine is feasible as immunization of pigs with some live attenuated ASFV vaccine candidates can confer protection, but safety concerns and virus scalability are challenges that must to be addressed. Identification of protective ASFV antigens is needed to inform the development of efficacious subunit vaccines.MethodsIn this study, replication-incompetent adenovirus-vectored multicistronic ASFV antigen expression constructs that covered nearly 100% of the ASFV proteome were generated and validated using ASFV convalescent serum. Swine were immunized with a cocktail of the expression constructs, designated Ad5-ASFV, alone or formulated with either Montanide ISA-201ā¢ (ASFV-ISA-201) or BioMizeĀ® adjuvant (ASFV-BioMize).ResultsThese constructs primed strong B cell responses as judged by anti-pp62-specific IgG responses. Notably, the Ad5-ASFV and the Ad5-ASFV ISA-201, but not the Ad5-ASFV BioMizeĀ®, immunogens primed significantly (p < 0.0001) higher anti-pp62-specific IgG responses compared with Ad5-Luciferase formulated with Montanide ISA-201ā¢ adjuvant (Luc-ISA-201). The anti-pp62-specific IgG responses underwent significant (p < 0.0001) recall in all the vaccinees after boosting and the induced antibodies strongly recognized ASFV (Georgia 2007/1)-infected primary swine cells. However, following challenge by contact spreaders, only one pig nearly immunized with the Ad5-ASFV cocktail survived. The survivor had no typical clinical symptoms, but had viral loads and lesions consistent with chronic ASF.DiscussionBesides the limited sample size used, the outcome suggests that in vivo antigen expression, but not the antigen content, might be the limitation of this immunization approach as the replication-incompetent adenovirus does not amplify in vivo to effectively prime and expand protective immunity or directly mimic the gene transcription mechanisms of attenuated ASFV. Addressing the in vivo antigen delivery limitations may yield promising outcomes
Biochemical Detection of Esterases in the Adult Female Integument of Organophosphate-Resistant Boophilus Microplus
Membrane Charge and Curvature Determine Interaction with Acyl-CoA Binding Protein (ACBP) and Fatty Acyl-CoA Targeting ā
A study of the systematics of Theileria spp. based upon small-subunit ribosomal RNA gene sequences
Persistence of replication-incompetent adenovirus in cattle.
<p>Viable recombinant adenovirus inoculated intradermally is only recoverable within three days. Presence of adenovirus rescued from tissue samples of four steers at defined time points was tracked by immunocytometric analysis of HEK-293A cells. Representative data from one steer is shown: A, D, G, J, and M are positive controls at 24 hr., 48 hr., 72 hr., day 7, and day 21, respectively. B, E, H, and K, are skin biopsies taken from the inoculation sites on the neck of the steers at 24 hr., 48 hr., 72 hr., and day 7, respectively, whereas C, F, I, and L, are cognate control skin biopsies taken concurrently from the flank. N and O are draining lymph node and spleen samples, respectively, collected three weeks post-inoculation.</p
Clinical manifestations post-challenge.
<p>A) Mean rectal temperature fluctuation; and B) Mean change ratios of white blood cell counts in the vaccinated and negative control groups post-challenge. Asterisks denote statistically significant differences as compared to the negative controls. *P<0.05; **P<0.01 and ***P<0.001.</p
Validation of mosaic antigens using BVDV-specific T-cells.
<p>Authenticity of T-cell epitopes in the mosaic BVDV antigens was validated by proliferation assay using PBMCs from a BVDV-1 & 2 hyper-immune steer [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0170425#pone.0170425.ref059" target="_blank">59</a>]. The data shown is minus background counts from negative control (media alone) treatment. The asterisks denote a statistically significant difference (P<0.01) between the proliferation induced by the N<sup>pro</sup>E2<sup>1-3</sup>, NS<sup>2-31</sup> and the NS<sup>2-32</sup> antigens and both whole killed viruses BVDV-1b and BVDV-2. This outcome is representative of assays conducted using PBMCs from other BVDV immune steers.</p