Evaluation of protection in a mouse model after vaccination with Mycobacterium avium subsp. paratuberculois protein cocktails

Abstract

Whole-cell vaccines successfully reduce signs of clinical disease and fecal shedding of Mycobacterium avium subsp. paratuberculosis (MAP), however, these vaccines have some limitations. The present study was conducted to identify MAP proteins that might be candidates for the development of an improved vaccine. MAP proteins were screened for immunogenicity in naturally infected cattle and selected based upon reactivity in the interferon-γ (IFN-γ) and Western blot assays. Proteins (MAP1087, MAP1204, MAP1272c, and MAP2077c) were arrayed into 4 overlapping cocktails containing 3 proteins each. The efficacy of the proteins within these cocktails as vaccine candidates was evaluated by subcutaneous immunization of mice, followed by challenge with live, virulent MAP. All MAP protein cocktails significantly reduced the recovery of live MAP from the ileum, while cocktails 1 and 3 reduced colonization in the liver. No significant differences were seen in the mesenteric lymph node or spleen, however, cocktail 1 reduced viable MAP in the mesenteric lymph node compared to other treatments. Stimulation of splenocytes upregulated antigen-specific IFN-γ and IL-23 secretion in all treatment groups, regardless of vaccination. Interestingly, IL-4 was moderately downregulated for vaccinates compared to control infected mice. An increase in total CD25 expression was noted for 3 of the 4 vaccinate groups upon stimulation of splenocytes with a whole cell sonicate of MAP, with this effect becoming more significant within CD4CD25+ and CD8CD25+ subpopulations. The present study demonstrated that MAP proteins are useful as vaccine candidates to reduce MAP tissue burden.

Virulence and Immunity Orchestrated by the Global Gene Regulator sigL in Mycobacterium avium subsp. paratuberculosis

“…Following trypan blue dye enumeration for cell viability, splenocytes were cultured in duplicate in round-bottom 96-well tissue culture plates with 1 ϫ 10 6 cells/well containing 100 U/ml human interleukin 2 (IL-2) (BD Biosciences) (26). Cells were restimulated in vitro with medium alone or medium supplemented with 10 g/ml Johnin purified protein derivative (PPDj) (USDA-National Veterinary Services Laboratory, Ames, IA) for 48 and 72 h at 37°C with 5% CO 2 (27). Cell supernatants were collected and levels of secreted cytokines were analyzed by mouse cytokine enzymelinked immunosorbent assay (ELISA) kit according to the manufacturer's instructions (BioLegend, San Diego, CA).…”

Molecular and histopathological evaluation of the efficacy of Saponifiedand Freund’s incomplete adjuvanted paratuberculosiskilled vaccine in murine model

“…Killed MAP vaccine was prepared using two different adjuvants, FIA and saponin (whole bacterin-QS21) as per Sigurdsson and co-workers (Sigurdsson, 1960) with suitable modifications to make killed saponified vaccine and FIA vaccine of MAP. Final concentration was adjusted in such a way that each ml contains 5 mg dry weight of bacteria (5 mg/ml) and a dose of 0.03ml containing 150µg of the antigen is given to each mice by S/C route (Stabel et al, 2012).…”

Therapeutic Potential of a Candidate relA Deletion Mutant Vaccine of Mycobacterium Avium Subsp. Paratuberculosis

“…paratuberculosis (Map) that induces sterile immunity against Johne's disease (JD) or prevents disease progression. These approaches have included development of killed whole vaccines, live attenuated, DNA and subunit vaccines and tests in different animal species (Bull et al 2007;Park et al 2008;Juste et al 2009;Kathaperumal et al 2009;Scandurra et al 2010;Bastida and Juste 2011;Stabel et al 2012;Faisal et al 2013). Although none of the vaccines induced sterile immunity they did slow disease progression and demonstrated that development of a vaccine that induces sterile immunity is feasible (reviewed in (Rosseels and Huygen 2008).…”