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Measurement of in vitro specific-antibody synthesis
1National Cancer Institute, Bethesda, Maryland, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
This study details a protocol to measure human lymphocyte responses to specific antigens like influenza virus. The method involves isolating cells, stimulating them, and quantifying antibody production using enzyme-linked immunosorbent assay (ELISA).
Area of Science:
- Immunology
- Cellular Biology
Background:
- Quantifying antigen-specific immune responses is crucial for understanding immunity.
- Human lymphocyte function assays require precise methodologies for reliable results.
Purpose of the Study:
- To provide a detailed protocol for eliciting and quantifying antigen-specific antibody responses from human lymphocytes.
- To establish a standardized method for immunological research using enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Isolation of mononuclear cells using Ficoll-Hypaque density gradient centrifugation.
- Treatment of cells to remove cytophilic antibodies prior to antigen stimulation.
- Stimulation of lymphocytes with specific antigens (e.g., influenza virus) or mitogens.
- Incubation for 12 days followed by supernatant harvesting for antibody quantification via ELISA.
Main Results:
- The protocol successfully elicits antigen-specific responses in human lymphocytes.
- Enzyme-linked immunosorbent assay (ELISA) effectively quantitates the antibody production.
Conclusions:
- This protocol offers a reliable method for assessing antigen-specific lymphocyte-mediated antibody responses.
- The described technique is adaptable for studying responses to various antigens and mitogens, aiding immunological research.
