Gamma-irradiated and nonirradiated Eimeria tenella sporozoites exhibit differential uracil uptake and expression of a 7- to 10-kDa metabolic antigen.
Eimeria tenella sporozoites were exposed in the oocyst form to either an optimum (15 kRad) or a high (25 kRad) dose of gamma irradiation and used to infect cultured chicken embryo fibroblasts (CEF). The sporozoite-infected CEF monolayer was pulsed at time of infection or 24 hr postinfection with [3H]uracil and harvested 24 hr later to measure sporozoite metabolic activity. Sporozoites exposed to either 0 or 15 kRad gamma irradiation incorporated similar (P > 0.05) amounts of [3H]uracil during the first and second 24-hr periods after infection. However, there was a significant decrease (P < 0.05) in [3H]uracil uptake by 25 kRad-exposed sporozoites compared to nonirradiated and 15 kRad-irradiated sporozoites. Indirect immunofluorescence (IFA) staining of E. tenella sporozoite-infected CEFs using monoclonal antibodies (MAb) specific for somatic or "metabolic" antigens showed that gamma irradiation also affected the release of intracellular metabolites. Regardless of irradiation dose, extracellular sporozoites exhibited similar intensity of immunofluorescence when stained with either somatic antigen- or metabolic antigen-reactive MAb. Also, somatic antigen expression was similar for intracellular parasites irrespective of radiation dose. However, metabolic 7- to 10-kDa antigen expression by 25 kRad-irradiated sporozoites was markedly reduced compared to nonirradiated or 15 kRad-irradiated intracellular sporozoites. These results were corroborated by immunostaining sporozoite/CEF protein-impregnated Immobilon membrane with somatic or metabolic 7- to 10-kDa antigen-reactive MAb. These findings may indicate that the metabolic 7- to 10-kDa antigen is involved in protective immunity elicited by nonirradiated and/or 15 kRad-irradiated E. tenella sporozoites.
Jenkins
M C
MC
Parasite Immunobiology Laboratory, LPSI, Agricultural Research Service, USDA, Beltsville, Maryland 20705, USA.
Chute
M B
MB
Danforth
H D
HD
Lillehoj
H S
HS
D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis*; D002478:Cells, Cultured; D002642:Chick Embryo; D016786:Eimeria tenella / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects*; D005347:Fibroblasts / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D005720:Gamma Rays; D015151:Immunoblotting; D015800:Protozoan Proteins / Q000032:analysis / Q000276:immunology; D014498:Uracil / Q000378:metabolism*
D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D014498:Uracil