Vaccination with recombinant adenovirus expressing peste des petits ruminants virus-F or -H proteins elicits T cell responses to epitopes that arises during PPRV infection

Abstract

Peste des petits ruminants virus (PPRV) causes an economically important disease that limits productivity in small domestic ruminants and often affects the livestock of the poorest populations in developing countries. Animals that survive PPRV develop strong cellular and humoral responses, which are probably necessary for protection. Vaccination should thus aim at mimicking these natural responses. Immunization strategies against this morbillivirus using recombinant adenoviruses expressing PPRV-F or -H proteins can protect PPRV-challenged animals and permit differentiation of infected from vaccinated animals. Little is known of the T cell repertoire these recombinant vaccines induce. In the present work, we identified several CD4+ and CD8+ T cell epitopes in sheep infected with PPRV. We also show that recombinant adenovirus vaccination induced T cell responses to the same epitopes, and led to memory T cell differentiation. T cells primed by these recombinant adenovirus vaccines expanded after PPRV challenge and probably contributed to protection. These data validate the use of recombinant adenovirus expressing PPRV genes as DIVA strategies to control this highly contagious disease.Electronic supplementary materialThe online version of this article (10.1186/s13567-017-0482-x) contains supplementary material, which is available to authorized users.

Bovine Herpesvirus-4-Based Vector Delivering Peste des Petits Ruminants Virus Hemagglutinin ORF Induces both Neutralizing Antibodies and Cytotoxic T Cell Responses

“…For responses to PPRV, splenocytes were cultured overnight with BEI-inactivated PPRV (Nig’75) ( 28 ). To assess responses to PPRV-H murine T cell epitopes H5 (H(551-559) YFYPVRLNF) and H9 (H(427-441) ITSVFGPLIPHLSGM) ( 29 ), splenocytes were expanded in vitro for 1 week with 10 µg/ml peptide before measuring IFN-γ responses. For intracellular IFN-γ measurements, cells were cultured at 10 6 cells per well in the presence of different stimuli (peptide or PPRV) overnight before the addition of 10 µg/ml brefeldin-A (Sigma) for the last 5 h of incubation.…”

“…Gating strategy is described in Ref. ( 29 ). Gating for positive IFN-γ positive events was set using isotype and fluorescence minus one channel controls.…”

Virus‐induced autophagic degradation of STAT 2 as a mechanism for interferon signaling blockade

“…Cells were then permeabilized and intracellular staining performed as described in [69]. Cells were then permeabilized and intracellular staining performed as described in [69].…”

“…For flow cytometry analysis of STAT2 expression, Vero cells were infected with BTV-8 at the indicated MOI and fixed after 24 h with 4% PFA for 15 min. Cells were then permeabilized and intracellular staining performed as described in [69]. Cells were stained with anti-STAT2 antibody and secondary Alexa Fluor 647-conjugated anti-rabbit IgG antibody (Thermo Fisher).…”

Comparative Analysis of BTS-34 and Vero-76 Cell lines for Isolation of Peste des Petits Ruminants (PPR) Virus

“…The disease is caused by a morbillivirus (PPRV) belonging to the family Paramyxoviridae (Chowdhury et al, 2014) and is characterized by high fever, ocular and nasal discharges; oral erosion, bronchopneumonia, gastroenteritis, necrotic stomatitis, and can lead to death. The morbidity and mortality rates can reach upto 100% (Hailat et al, 2018) and 50-90% respectively in susceptible populations (Rojas et al, 2017).…”