Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 1, 2007
Publication Date: June 1, 2007
Citation: Scott, T.R., Lillehoj, H.S. 2007. Monoclonal antibodies against chicken interleukin-6. Veterinary Immunology and Immunopathology 114:173-177.
Interpretive Summary: Study of host-pathogen interactions is important in order to develop logical vaccines for many poultry diseases. In this paper, ARS scientist collaborated with a scientist at Clemson University to develop several mouse hybridomas which secrete monoclonal antibodies that detect chicken interleukin 6 (IL-6). IL-6 is a pleiotropic, proinflammatory cytokine, and its presence in serum of infected chickens indicates ongoing inflammatory host response. This is the first report documenting the development of antibodies which are specific for chicken IL-6 and availability of these antibodies will facilitate basic and applied research in disease of poultry.
Monoclonal antibodies (mAb) were produced against a recombinant (r) chicken interleukin-6 (IL-6). Eight mAbs that were produced were tested for isotype; ability to inhibit recombinant forms of chicken (ch), human (h) and murine (m) IL-6; and recognition of rchIL-6 by Western immunoblotting. The mAb isotypes were represented by IgG1 (one), IgG2a (six) and IgG2b (one). In an initial mouse B9 hybridoma cell bioassay with rmIL-6, four mAbs against rchIL-6 effectively inhibited the activity of rmIL-6. Further bioassays with the four mAbs at varying concentrations showed that two of these mAbs (1.20.7 and 1.26.4) were quite effective at inhibiting rmIL-6. Recombinant forms of ch, h and mIL-6 were all tested in a bioassay with the most potent inhibiting mAb (1.26.4), and this mAb was effective in inhibiting the activity of all three recombinant IL-6 proteins. Western immunoblotting revealed identification of the original IL-6 immunogen used for mAb production. Based upon inhibition of IL-6 activity in a standard bioassay and IL-6 recognition by Western immunoblotting, mAb 1.26.4 was judged the most useful antibody for future studies and applications.