Establishment of swine interleukin-6 sandwich ELISA.
We established a sandwich enzyme-linked immunosorbent assay (ELISA) for swine interleukin-6 (SwIL-6), which was applied for detection of SwIL-6 in vitro and in vivo. Anti-SwIL-6 rabbit- and goat-polyclonal antibodies, and monoclonal antibody (mAb) were prepared, conforming that all of the antibodies were reactive with recombinant SwIL-6 by Western blotting and indirect ELISA. A sandwich ELISA was developed using the mAb as a capture antibody and biotinylated goat-polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA for rSwIL-6 was 49pg/ml and did not show cross-reactivity with swine IL-1b, IL-4, IL-8, IL-18, IL-12, and IFN-g. Using the ELISA, SwIL-6 was detected in culture medium of the monocytes stimulated with PHA-P and PMA, and the plasma or the bronchoalveolar lavage fluid (BALF) of pigs experimentally infected with Actinobacillus pleuropneumoniae or Mycoplasma hyopneumoniae. This ELISA for SwIL-6 may be useful for understanding the role of this cytokine in various swine diseases.
Nuntaprasert
A
A
Laboratory of Animal Research Center, Institution of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai Minato-ku, Tokyo 108-8639, Japan.
Mori
Y
Y
Tsukiyama-Kohara
K
K
Kai
C
C
D000189:Actinobacillus Infections / Q000276:immunology / Q000662:veterinary; D016977:Actinobacillus pleuropneumoniae / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015850:Interleukin-6 / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D045705:Mycoplasma hyopneumoniae / Q000276:immunology; D045729:Pneumonia of Swine, Mycoplasmal / Q000276:immunology; D013552:Swine / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015850:Interleukin-6