Abstract
Chickens were immunized subcutaneously with an Eimeria recombinant profilin protein plus Montanide™ ISA 70 VG (ISA 70) or Montanide™ ISA 71 VG (ISA 71) water-in-oil adjuvants, or with profilin alone, and comparative RNA microarray hybridizations were performed to ascertain global transcriptome changes induced by profilin/ISA 70 vs. profilin alone and by profilin/ISA 71 vs. profilin alone. While immunization with profilin/ISA 70 vs. profilin alone altered the levels of more total transcripts compared with profilin/ISA 71 vs. profilin alone (509 vs. 296), the latter was associated with a greater number of unique biological functions, and a larger number of genes within these functions, compared with the former. Further, canonical pathway analysis identified 10 pathways that were associated with genes encoding the altered transcripts in animals immunized with profilin/ISA 71 vs. profilin alone, compared with only 2 pathways in profilin/ISA 70 vs. profilin alone. Therefore, ISA 71 was selected as a candidate adjuvant in conjunction with profilin vaccination for in vivo disease protection studies. Vaccination with profilin/ISA 71 was associated with greater body weight gain following E. acervulina infection, and decreased parasite fecal shedding after E. maxima infection, compared with profilin alone. Anti-profilin antibody levels were higher in sera of E. maxima- and E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. Finally, the levels of transcripts encoding interferon-γ, interleukin (IL)-2, IL-10, and IL-17A were increased in intestinal lymphocytes from E. acervulina-, E. maxima-, and/or E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. None of these effects were seen in chickens injected with ISA 71 alone indicating that the adjuvant was not conferring non-specific immune stimulation. These results suggest that profilin plus ISA 71 augments protective immunity against selective Eimeria species in chickens.
Highlights
Avian coccidiosis is one of the most costly infectious diseases affecting the commercial poultry industry [1]
This study demonstrated that chickens immunized with profilin plus ISA or profilin plus ISA exhibited spleen lymphocyte transcriptome changes, compared with immunization with profilin alone, that were consistent with alteration of levels of mRNAs encoded by genes belonging to the IPA database category ‘‘Disease and Disorder’’
Weight gains and parasite shedding were equal in the ISA 71 VG (ISA 71) alone vs. PBS negative control groups, and intraepithelial lymphocytes (IELs) transcript levels were generally equivalent in the ISA 71 only vs. profilin alone groups, indicating that the adjuvant itself was not responsible for these effects
Summary
Avian coccidiosis is one of the most costly infectious diseases affecting the commercial poultry industry [1]. Coccidia that infect chickens include Eimeria acervulina, E. tenella, E. maxima, E. brunetti, E. necatrix, E. praecox, and E. mitis. These apicomplexan protists invade cells of the intestinal epithelium, evoking necrotic tissue destruction and resulting in reduced body weight gain in broilers, decreased egg production in layers, and fecal shedding of viable parasites that re-infect susceptible animals upon ingestion. The basis of using live coccidia vaccines involves a continuous excretion/reingestion cycle of an initial low dose of parasites, which progressively induces protective flock immunity. Novel approaches are needed to more effectively control coccidiosis in commercial poultry flocks
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