A recombinant Eimeria protein inducing interferon-γ production:: Comparison of different gene expression systems and immunization strategies for vaccination against coccidiosis

被引:97
作者
Lillehoj, HS
Choi, KD
Jenkins, MC
Vakharia, VN
Song, KD
Han, JY
Lillehoj, EP
机构
[1] USDA, BARC E, Livestock & Poultry Sci Inst, Immunol & Dis Resistance Lab, Beltsville, MD 20705 USA
[2] Virginia Maryland Reg Coll Vet Med, College Pk, MD 20742 USA
[3] Ctr Agr Biotechnol, College Pk, MD 20742 USA
[4] Seoul Natl Univ, Coll Agr & Life Sci, Dept Anim Sci & Technol, Suwon 441744, South Korea
[5] Dexall Biomed Labs Inc, Gaithersburg, MD 20879 USA
关键词
Eimeria; coccidiosis; recombinant protein; interferon-gamma; vaccination; chickens;
D O I
10.2307/1592553
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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 recombinant 3-1E cDNA expressed in Escherichia coli produced a 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 Sm insect cells and a 2D-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)-gamma production upon stimulation with the recombinant 3-1E protein, indicating that the protein activates cell-mediated immunity during coccidiosis. Immunization of chickens with either the E. coli- or Sf9-expressed recombinant 3-1E 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-gamma 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.
引用
收藏
页码:379 / 389
页数:11
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