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Possible 110 kDa receptor for interleukin 2 in the chicken
1Department of Biomedical and Environmental Health Sciences, School of Public Health, University of California, Berkeley 94720.
Insights
Researchers developed monoclonal antibodies to identify chicken lymphokine receptors on activated lymphocytes. One antibody specifically targets a 110 kDa surface antigen, potentially analogous to the human interleukin 2 receptor gamma subunit.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Chicken lymphoblasts are activated lymphocytes crucial for immune responses.
- Lymphokines mediate communication between immune cells, but their receptors in chickens are not well characterized.
Purpose of the Study:
- To produce and characterize monoclonal antibodies (mAbs) for identifying chicken lymphokine receptors.
- To investigate the function and molecular properties of a specific lymphokine receptor on chicken lymphoblasts.
Main Methods:
- Generation of chicken lymphoblasts via concanavalin A (Con A) stimulation.
- Production of mAbs by immunizing mice with activated lymphoblasts.
- Selection of specific mAbs using cellular enzyme immunoassay (EIA) and immunofluorescence staining.
- Assessment of mAb inhibitory activity on lymphokine function.
- Immunoprecipitation to determine the molecular weight of the target antigen.
Main Results:
- A specific mAb was generated that strongly reacted with lymphoblasts but weakly with resting cells, erythrocytes, and bursal cells.
- This mAb inhibited lymphokine activity in conditioned media from activated spleen cells.
- Immunoprecipitation identified a lymphoblast surface antigen of approximately 110 kDa.
Conclusions:
- A novel monoclonal antibody successfully identified a specific lymphokine receptor on chicken lymphoblasts.
- The identified 110 kDa receptor may be functionally and structurally analogous to the interleukin 2 receptor gamma subunit in mammals.
- This research provides a tool for further studying chicken immune cell activation and lymphokine signaling.
Abstract:
Chicken lymphoblasts were generated from spleen cells which had been incubated with concanavalin A (Con A) for 48 h. Monoclonal antibodies (mAbs) were produced by immunizing mice with 48-h Con A lymphoblasts. These antibodies were used to identify chicken lymphokine receptors on Con A-activated lymphocytes. The cellular enzyme immunoassay (EIA), with lymphoblasts and spleen cells as the target cells, was used to select for specific mAbs. One mAb was found by EIA to have a strong response against the lymphoblasts but weak against resting spleen cells. By immunofluorescence staining, it reacted strongly with lymphoblasts and weakly with resting spleen, erythrocytes, and bursal cells. This mAb was also shown to inhibit the lymphokine activity present in conditioned medium that was collected from 24-h cultures of Con A-activated spleen cells. By immunoprecipitation, this mAb precipitated a lymphoblast surface antigen with a molecular weight of about 110 kDa. This receptor may be analogous to the putative interleukin 2 gamma subunit recently described in human and mouse cell lines.
