Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood.

TLDR

Conventional T‑cell enumeration methods are laborious, taking up to two days and requiring at least 20 mL of blood. The study presents a simple, rapid method to determine T‑cell subclasses in buffy coat or whole blood. The method uses flow cytometry to separate lymphocytes from other leukocytes by light‑scattering, then enumerates T‑cell subclasses via immunofluorescence with monoclonal antibodies against surface differentiation antigens. The technique requires only 50 µL of whole blood per antibody and delivers results in under two hours.

Abstract

A simple and rapid method for the determination of human T lymphocyte subclasses in buffy coat preparations or whole blood is described. This technique uses flow cytometry to distinguish lymphocytes from other leukocytes on the basis of their light-scattering properties. Lymnphocyte subclasses were enumerated by cellular immunofluorescence; the immunofluorescent signals were produced by monoclonal antibodies to surface differentiation antigens on T cells. Conventional techniques of enumerating T lymphocyte subclasses entail time-consuming (up to 2 days) density gradient and E rosette enrichement, and require at least 20 ml of blood. The method described here uses as little as 50 microliters of whole blood for each antibody tested and produces results within in 2 hr.