Measurement of polyclonal immunoglobulin synthesis using the reverse plaque technique.

Area of Science:

• Immunology
• Cell Biology

Background:

• Immunoglobulin (Ig)-secreting cells are vital for adaptive immunity.
• Accurate quantification of these cells is essential for immunological studies.

Purpose of the Study:

• To describe the reverse hemolytic plaque assay (RHPA) for measuring Ig-secreting cells.
• To provide a detailed protocol for conducting the RHPA.

Main Methods:

• The RHPA involves incubating test cell populations with protein A-sensitized sheep red blood cells (SRBC), complement, and anti-Ig antibody.
• Incubation is followed by scoring of resulting hemolytic plaques.
• A support protocol details the preparation of specialized plaquing chambers.

Main Results:

• The RHPA effectively measures the number of Ig-secreting cells in a population.
• The assay can be applied to diverse cell sources, including peripheral blood mononuclear cells and tonsil cells.

Conclusions:

• The reverse hemolytic plaque assay is a valuable and effective technique for enumerating Ig-secreting cells.
• This method facilitates the study of humoral immunity and antibody production in various biological samples.