Cytokine expression by high‐density human lymphocytes

• Published in: Immunology 1996-03, Vol.87 (3), p.408-413
• Main Authors: SEWELL, W. A., BERGER, M. F., SKIPSEY, L. J., COOLEY, M. A., WARREN, H. S.
• Format: Article
• Language: English
• Subjects: Base Sequence; Cell Separation; Cytokines - genetics; Cytokines - metabolism; DNA Primers - genetics; Humans; Interferon-gamma - metabolism; Interleukin-2 - metabolism; Interleukin-3 - metabolism; Interleukin-4 - metabolism; Interleukin-5 - metabolism; Leukocyte Common Antigens - analysis; Leukocytes, Mononuclear - immunology; Lymphocyte Activation; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Messenger - metabolism; T-Lymphocytes - immunology
• ISSN: 0019-2805; 1365-2567
• DOI: 10.1046/j.1365-2567.1996.501572.x

T lymphocytes spend much of the time as small non‐cycling cells. To determine the pattern of cytokine expression in such resting cells, they were purified from human peripheral blood mononuclear cells (PBMC) on the basis of high buoyant density. The cells were stimulated and cytokine mRNA expression was assessed by reverse transcription–polymerase chain reaction (RT‐PCR). Expression of interleukin‐2 (IL‐2), IL‐3 and interferon‐γ (IFN‐γ) was similar in high‐density lymphocytes and in unfractionated PBMC. In contrast, the high‐density lymphocytes expressed less IL‐4 than PBMC, and little or no IL‐5. Because a substantial minority of the high‐density lymphocytes was CD45RO+, the presence of this marker was not an indicator of the ability to express IL‐4 and IL‐5. In the high‐density lymphocytes, IFN‐γ expression was confined to the CD45RO+ fraction, whereas IL‐2 was expressed by both CD45RO+ and CD45RO− subsets. To assess whether high‐density lymphocytes could give rise to cells with a broader range of inducible cytokine expression, they were activated and then restimulated between 10 and 22 days of culture. Cells derived from both the CD45RO+ and CD45RO− fractions of high‐density lymphocytes expressed IL‐5 after restimulation. Thus the high‐density lymphocyte population has the potential to acquire a broader range of inducible cytokine expression.