scope:

This document provides guidance for the performance of single cell immune response assays in the clinical areas including infectious disease (especially human immunodeficiency virus (HIV)), cancer, transplantation, autoimmune disease, and allergy. It has recently been recognized that there is a need to identify and validate markers of immune competency that can be used in clinical trials and for approval of immune therapy by regulatory agencies. This guideline will therefore focus on specific assays to evaluate CD4 and CD8 T-cell function. The assays in this document include methods for measurement of specific antigen-induced responses by flow cytometric determination of intracellular cytokines and other functional molecules (e.g., perforin) and by ELISPOT assay, as well as MHC tetramer technology for the quantification of antigen-specific CD4 and CD8 T cells. The document also covers issues such as specimen collection and transport, sample preparation, and quality control.

The guideline will provide laboratory methods for the growing field of immune-based therapy. It will provide guidance to pharmaceutical manufacturers in the laboratory evaluation of new products before submission to regulatory agencies. It will also be a valuable resource for academic investigators coordinating research between laboratories participating in multicenter trials. Overall, this guideline establishes consensus methods for a rapidly evolving field of single cell immune functional assays.