A NOVEL SENSITIVE ASSAY TO DEFINE IMMUNE STATUS USING SHORT-TERM PERIPHERAL-BLOOD DERIVED CELL-CULTURE AND DUAL-COLOR FLOW-CYTOMETRY

In this study we describe a novel and highly sensitive in vitro system to determine the functionality of immune cells based on short term cu lture of peripheral blood derived mononuclear cells (PBMCs) and subseq uent analysis of cellular proliferation and surface marker expression by automated dual-color flow cytometry. The standardized mild stimuli introduced into the culture system by supplemented medium (containing exogenous interleukin-2 (IL-2), and fetal bovine serum (FBS)) allow a more physiological interaction of the different cell subsets contained in PBMCs (including CD14(+) accessory cells) than other methods that are based on potent and harsh cell activators, such as phytohemaggluti nin (PHA) or anti-CD3 antibodies. Measurement of T-cell proliferation and cell surface marker (CD3, C25, CD26, CD71, HLA-DR) analysis reveal ed that activation response capacity in our assay depends on both the status of the obtained cells and their ability to interact in culture with CD14+ cells. This in vitro assay proved to be very sensitive in d etecting changes in the status of T-cell activation and proliferation capacity, and avoid the use of radioactive reagents. (C) 1998 Elsevier Science B.V. All rights reserved.