The delayed type hypersensitivity assay using protein and xenogeneic cell antigens

The delayed-type hypersensitivity (DTH) assay has a lengthy history in immunotoxicity testing since it was one of the original functional assays included in the National Toxicology Program (NTP) immunotoxicology test panel. Based on NTP data analysis, the DTH assay is among the most predictive immunotoxicity tests when included with at least two other immune parameters. The DTH assay has the advantage of being: (1) a useful measure of cell-mediated immunity, (2) an in vivo assay where there is less opportunity for ex vivo confounders and (3) a clinically significant human correlate to the tuberculin test. Disadvantages of the DTH assay are that it is potentially labor-intensive to perform, it is somewhat resistant to automation and, when compared with the cyctotoxic T lymphocyte (CTL) assay, it is a relatively crude measurement. However, some groups have been attempting to address the limitations of the DTH assays (see Note 1).The assay is related to the contact hypersensitivity response (CHR), which is covered in another chapter. The DTH response has been used as an indicator of cell-mediated immune status and is dependent upon both T helper 1(Th1)-driven responses as well as cell recruitment and chemotaxis to a local site. As a result, the DTH functional response may be influenced by disruption of either Th1-driven, antigen-dependent T cell development or mobilization of sensitized T cells to a local site. The present chapter describes four common protocols with consideration restricted to protein and xenogeneic cell immunogens.