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United States Department of Agriculture

Agricultural Research Service

Title: Production and Characterization of Monoclonal Antibodies Detecting Chicken Interleukin-2 and the Development of An Antigen Capture Enzyme-Linked Immunosorbent Assay

item Miyamoto, Tadashi - USDA:ARS:ANRI:PBESL
item Lillehoj, Hyun
item Sohn, Eun - USDA:ARS:ANRI:PBESL
item Min, Wongi - USDA:ARS:ANRI:PBESL

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 20, 2001
Publication Date: N/A

Interpretive Summary: For many poultry diseases, understanding complex interactions between host immune system and microbes is critical to develop a novel control strategy. Recent studies have clearly shown that cell-mediated immunity (CMI) is an important aspect of protective host response to microbial pathogens and interleukin-2 (IL-2) is an essential cytokine that mediates many fundamental immune processes in mammals and chickens. IL-2 is a powerful growth factor for a variety of cell types and plays a key role in many aspects of host immune response. Studies from this laboratory indicate that chicken IL-2 has practical importance in potentially enhancing immune responses to vaccines and infectious agents in livestock and poultry. Therefore, ARS scientists developed many mouse monoclonal antibodies which specifically identify chicken IL-2 to develop a simple, rapid, and highly reliable ELISA in this study. Availability of sensitive ELISA will enhance the ability of US poultry industry to assess the immune response of poultry in many disease processes

Technical Abstract: Eleven monoclonal antibodies (mAbs) which are specific for chicken interleukin-2 (chIL-2) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting and neutralizing assays. These mAbs were used to develop a mAb-based antigen capture ELISA specific for chicken IL-2 detection. Anti- IL-2 mAbs bound specifically to E. coli-derived rchIL-2 in ELISA and identified a 16-kDa IL-2 polypeptide band in Western blot. Several mAbs were shown to neutralize the biological activities of both rchIL-2 and native chicken IL-2. To develop a sensitive ELISA for the detection of chicken IL-2, an antigen capture ELISA was developed using the mAb chIL-2/16 as the antigen capture antibody and rabbit anti-IL-2 peptide antibody as the detection antibody. Using the mAb-based antigen capture ELISA, significant correlation between the level of IL-2 detected in bioassays and in ELISA was observed. These results showed that the mAb-based antigen capture ELISA is less time-consuming and reliable compare to a conventional IL-2 bioassay for chicken IL-2. These neutralizing mAbs will facilitate basic immunobiological studies of the role of IL-2 in normal and disease states in chickens.

Last Modified: 4/22/2015
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