S-Space College of Agriculture and Life Sciences (농업생명과학대학) Dept. of Food and Animal Biotechnology (식품·동물생명공학부) Journal Papers (저널논문_식품·동물생명공학부)
A recombinant Eimeria protein inducing interferon-γ production: comparison of different gene expression systems and immunization strategies for vaccination against coccidiosis
- Lillehoj, Hyun S.; Choi, Kang D.; Jenkins, Mark C.; Vakharia, Vikram N.; Song, Ki D.; Han, Jae Yong; Lillehoj, Erik P.
- Issue Date
- Avian Diseases, vol.44 no.2, pp. 379-389
- SUMMARY. A rabbit antiserum against an 18- to 27-kD native protein fraction (F3)
from Eimeria acervulina merozoites identified a cDNA (3-1E) containing a 1086-base pair
insertion with an open reading frame of 170 amino acids (predicted molecular weight,
18,523). The recombinant3 -1E cDNA expressedi n Escherichiaco li produceda 60-kD fusion
protein and a 23-kD protein after factor Xa treatment of the fusion protein. Both proteins
were reactive with the F3 antiserum by western blot analysis. A rabbit antiserum against a
synthetic peptide deduced from the amino acid sequence of the 3-1E cDNA reacted with a
27-kD recombinant 3-1E protein expressed in Sf9 insect cells and a 20-kD native protein
expressed by E. acervulina sporozoites and Eimeria tenella sporozoites and merozoites. By
immunofluorescence staining, a monoclonal antibody produced against the recombinant 3-
1E protein reacted with sporozoites and merozoites of E. acervulina, E. tenella, and Eimeria
maxima. Spleen lymphocytes from E. acervulina-immune chickens showed antigen-specific
proliferation and interferon (IFN)-y production upon stimulation with the recombinant 3-
1E protein, indicating that the protein activates cell-mediated immunity during coccidiosis.
Immunizationo f chickensw ith either the E. coli- or Sf9-expressedr ecombinant3 -1E protein
with adjuvant, or direct injection of the 3-1E cDNA, induced protective immunity against
live E. acervulina. Simultaneous injection of the recombinant 3-1E protein, or the 3-1E
cDNA, with cDNAs encoding chicken IFN-y or interleukin (IL)-2/15 further enhanced
protective immunity. These results indicate that the recombinant E. acervulina 3-1E cDNA
or its polypeptide product may prove useful as vaccines against avian coccidiosis.
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