Monoclonal antibodies reactive with chicken interleukin-17.
Chicken interleukin-17 (chIL-17) gene was previously characterized through cloning from a chicken intestinal expressed sequence tag (EST) cDNA library. To further investigate the biological properties of chIL-17, six monoclonal antibodies (mAbs) against a bacterially expressed chIL-17 recombinant protein were produced and their binding specificities characterized. Antibodies which were initially selected on the basis of their specific binding reactivity with recombinant chIL-17 in ELISA were further characterized by Western blot analysis. Monoclonal antibodies specific for chIL-17 identified 20 and 21kDa protein bands in the culture supernatant and cell lysate of CU205 cells. These mAbs also recognized specific bands for chIL-17 in the cell lysate from conconavalin A (Con A)-activated, but not from normal splenic lymphocytes. Furthermore, these mAbs detected a 16kDa protein in the lysate of CU205 cells treated with tunicamycin and stained an intracellular protein in CU205 cells in flow cytometric analysis. Together, these results indicate that these new mAbs are specific for chIL-17 and will be a useful tool for structural and immunological studies of IL-17 in poultry.
Yoo
Jeongmi
J
College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Korea.
Chang
Hong H
HH
Bae
Yeong H
YH
Seong
Chi-Nam
CN
Choe
Nong-Hoon
NH
Lillehoj
Hyun S
HS
Park
Jong-Hyeon
JH
Min
Wongi
W
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D002461:Cell Line, Transformed; D002645:Chickens / Q000235:genetics / Q000276:immunology*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D006825:Hybridomas / Q000276:immunology; D020381:Interleukin-17 / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D020381:Interleukin-17; D011994:Recombinant Proteins; D004247:DNA