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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.

Evaluation of Montanide™ ISA 71 VG Adjuvant during Profilin Vaccination against Experimental Coccidiosis