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

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- (IFN-) 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- 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

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

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- (IFN-) 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- 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

Genetic Association of Bovine Lymphocyte Antigen DRB3 Alleles with Immunological Traits of Holstein Cattle

The associations between alleles at the BoLA (bovine lymphocyte antigen) DRB3 locus and 20 indicator traits of innate and adaptive immunity were investigated. Periparturient Holsteins (n = 127) were genotyped at the BoLA DRB3 locus using polymerase chain reaction and restriction fragment length polymorphism. Twenty-two alleles were observed in the study population, and frequencies ranged from 21 to <1%. The same cattle were tested for a total of 20 innate and adaptive immunity traits, including lymphocyte response to mitogens (proliferative responses and Ig secretion), serum Ig, complement and conglutinin concentrations, total leukocyte count, and selected assays for neutrophil function. Models with gene substitution effects were used to investigate associations between BoLA DRB3 alleles and each of the immunological variables. Significant associations were found with 13 of the leukocyte functions tested. The number of immune parameters with significant associations with any allele ranged from 0 (with alleles DRB3.2*23 and DRB3.2*27) to 7 (with DRB3.2*8). The immunological parameter that had the most associations with alleles was serum IgG2 concentration with 6 alleles. One group of 4 alleles (representing 46% of the total allele frequency) was uniformly associated with increased IgM and complement and decreased mononuclear cell numbers. Thus, we demonstrated that the BoLA DRB3 genotype can influence measures of innate and adaptive immunity