ELISpot

ELISpot is a type of assay that focuses on the quantification and detection of immune cells which secrete specific cytokines. This assay is commonly used in the field of immunology and virology to study immune responses to infections, vaccines, and cancer.

History

The ELISpot assay was first developed in the early 1980s by Cecil Czerkinsky and his colleagues at the University of Gothenburg in Sweden. The initial application of the assay was to detect and quantify B cells secreting antigen-specific antibodies.

Principle

The principle of the ELISpot assay is based on the enzyme-linked immunosorbent assay (ELISA). It involves the capture of cytokines secreted by individual cells on a surface coated with specific antibodies. The captured cytokines are then detected using a second set of antibodies labeled with an enzyme. The enzyme catalyzes a color reaction at the site of cytokine capture, resulting in the formation of a visible spot. Each spot represents a single cytokine-secreting cell.

Procedure

The procedure of the ELISpot assay involves several steps. First, a surface (usually a microtiter plate) is coated with a specific antibody. The cells of interest are then added to the plate and incubated for a certain period of time. During this incubation, the cells secrete cytokines which are captured by the antibodies on the surface. After the incubation, the cells are removed and a second set of antibodies labeled with an enzyme is added. These antibodies bind to the captured cytokines. Finally, a substrate for the enzyme is added, which results in a color reaction at the site of cytokine capture. The spots are then counted using a specialized microscope or an automated reader.

Applications

The ELISpot assay has a wide range of applications in the field of immunology and virology. It is used to study immune responses to infections, vaccines, and cancer. It can also be used to detect and quantify specific types of immune cells, such as T cells and B cells.

Advantages and Limitations

The main advantage of the ELISpot assay is its high sensitivity and specificity. It allows the detection and quantification of individual cells secreting specific cytokines. However, the assay has some limitations. It requires specialized equipment and trained personnel. In addition, the results can be influenced by various factors, such as the quality of the antibodies and the conditions of the assay.

See Also

• Enzyme-linked immunosorbent assay
• Immunology
• Virology

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