Nirmala, R and Narayanan, P R
(2002)
Flowcytometry – A rapid tool to correlate functional activities of
human peripheral blood lymphocytes with their corresponding
phenotypes after in vitro stimulation.
BMC Immunology, 3 (9).
ISSN ESSN: 1471-2172
Abstract
Background: While dealing with mixed in vitro lymphocyte cultures one is faced with the problem
of relative contributions of different populations to the activity being studied. This is especially true
in the controversy relating to the contributions of lymphocyte sub-populations to the Lymphokine
Activated Killer (LAK) phenomenon. Flowcytometry can be used to highlight relative contributions
of lymphocyte subpopulations towards LAK activity without resorting to difficult purification
strategies. We set up long-term in vitro lymphocyte cultures, stimulated them with cytokines IL-2/
IL-12, recorded their phenotypic changes and cytotoxic activity against U-937 tumor targets.
Results: The results indicated that natural killer cells (NK) constituted the predominant
proliferating cell population in the cytokine stimulatedcultures. Flowcytometric evidence revealed
that CD56+ T cells contributed little to LAK activity against U937 target cells as compared to cells
with NK phenotype which were predominantly responsible for spontaneous killing of the tumor
targets. The two cytokines, IL-2 and IL-12, had an additive effect on cell proliferation and
spontaneous cytotoxicity.
Conclusion: Flowcytometry can be used to rapidly delineate phenotypic changes in immune cells
after stimulation and simultaneously correlate them with corresponding functional activity. This
approach may find application as a initial screening tool for studying different types of cells in mixed
cultures and their respective activities under stimulatory / inhibitory conditions.
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