Computer-assisted, quantitative cytokine enzyme-linked immunospot analysis of human immune effector cell function

Abstract

Originally developed for detecting antibody production from B lymphocytes, the enzyme-linked immunospot (ELISPOT) assay was later modified to assess cytokine production from various immune effector cells. Although the ELISPOT assay can detect antibody or cytokine production at the single-cell level, the visual counting of spots in a 96-well plate under a microscope makes this method unsuitable for handling large sample sizes. Here, we introduce a computer-assisted image analysis system to overcome this problem. This system makes the data analysis step of the ELISPOT assay convenient, objective, sensitive and suitable for handling large sample pools. Studies requiring lymphocyte proliferation assay, cytotoxic lymphocyte assay and precursor frequency assay can be conducted through the ELISPOT assay. This is demonstrated here using examples such as mixed lymphocyte allogeneic reactions and human immunodeficiency virus antigen-specific, cell-mediated immune responses.