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H5D12 monoclonal antibody identifies a 24-kD immunosuppressor factor produced by human chorio carcinoma cells
1Department of Obstetrics, Gynecology and Reproductive Sciences, University of Manitoba, Winnipeg, Canada.
Insights
Researchers identified a unique 24-kD embryo-associated immunosuppressor factor (EASF) secreted by chorio carcinoma cells. This EASF possesses immunosuppressive activity, potentially regulating reproductive functions.
Area of Science:
- Reproductive immunology
- Cell biology
- Cancer research
Background:
- Chorio carcinoma cells secrete factors that can influence the maternal immune response during pregnancy.
- Identifying these factors is crucial for understanding reproductive health and potential complications.
Purpose of the Study:
- To identify and characterize the embryo-associated immunosuppressor factor (EASF) secreted by chorio carcinoma cells.
- To utilize the EASF-specific monoclonal antibody H5D12 for this characterization.
Main Methods:
- Immunohistochemistry on chorio carcinoma and control cell lines using antibody H5D12.
- Affinity purification of EASF from cell culture media.
- Assessing immunosuppressive activity via lymphocyte proliferation assays.
- Determining molecular weight using metabolic labeling, immunoprecipitation, and SDS-PAGE.
Main Results:
- Antibody H5D12 strongly stained BeWo chorio carcinoma cells.
- Biosynthetic labeling identified EASF as a 24-kD molecule.
- EASF demonstrated significant immunosuppressive activity in bioassays.
- Control cell lines did not exhibit similar immunosuppressive factors or molecules.
Conclusions:
- Monoclonal antibody H5D12 successfully identified a 24-kD immunosuppressive factor secreted by chorio carcinoma cells.
- This factor, EASF, is unique and may play a key role in regulating reproductive functions.
Problem:
The purpose of this study was to identify and characterize embryo-associated immunosuppressor factor (EASF) secreted by chorio carcinoma cells with the help of EASF-specific monoclonal antibody H5D12 (raised against EASF purified from embryo growth media of in vitro fertilized human ova).
Method:
Paraffin-embedded slides of human chorio carcinoma as well as control cell lines were prepared, and immunohistochemistry was done by the avidin-biotin-peroxidase technique. EASF was affinity purified using H5D12-Sepharose 4B from culture media of cell lines and analyzed for immunosuppressive activity (by Concanavalin-A-induced lymphocyte proliferation assay) and molecular weight identity (by metabolic-labelling studies with 35S-methionine followed by immunoprecipitation and SDS-PAGE).
Results:
H5D12 showed intense immunostaining of BeWo chorio carcinoma cells. Biosynthetic labeling studies identifies this factor as a 24-kD molecule, and EASF bioassay indicates that this factor possesses immunosuppressive activity. No such immunosuppressive activity or similar molecules were identified when control cell lines were analyzed.
Conclusions:
Monoclonal antibody H5D12 recognizes a 24-kD factor with immunosuppressive activity that is secreted by chorio carcinoma cells, which suggests that this is a unique factor and may be one of the key regulators of reproductive functions.
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