Effects of hypoxia on interleukin-2 mRNA expression by T lymphocytes

A. L. Zuckerberg, L. I. Goldberg, Howard M Lederman

Research output: Contribution to journalArticle

Abstract

Objective: To investigate the effects of hypoxia on T-lymphocyte expression of IL-2 messenger RNA (mRNA), after cell activation with phorbol ester and ionomycin. Design: Prospective, controlled, cellular trial. Setting: University research laboratory. Subjects: EL4.6.1 cells, a murine T- cell lymphoma line. Interventions: Tissue culture media was deoxygenated and flushed continuously with 100% helium to maintain a PO2 of 30 to 40 torr (2 of 45 to 55 torr (>45 torr [>6.0 kPa]). The pH was maintained between 7.3 and 7.6. The media was inoculated with EL4 cells. Aliquots of cells were obtained at intervals and divided into two groups: an immediate group, stimulated immediately, and an overnight group that was returned to normal incubator conditions of 5% CO2/humidified room air for 18 hrs before stimulation. Measurements and Main Results: Gas tension, pH, cell count, and viability were determined for each aliquot. Cells were stimulated with phorbol myristate acetate and ionomycin for 4 hrs, at which time levels of interleukin-2 (IL-2) messenger RNA (mRNA) and gamma actin mRNA were measured by solution hybridization and enzyme immunoassay. The results were expressed as IL-2 mRNA/gamma actin mRNA ratio, normalized to baseline room air values. Cell viability and housekeeping functions (gamma actin mRNA expression) were unaffected by hypoxia. Cells exposed to a PO2 of 2 of >45 torr (>6.0 kPa). Conclusions: The regulation of IL-2 transcription in the T lymphocyte appears to be exquisitely sensitive to changes in oxygen tension. Exposure to a PO2 of

Original languageEnglish (US)
Pages (from-to)197-203
Number of pages7
JournalCritical Care Medicine
Volume22
Issue number2
StatePublished - 1994

Fingerprint

Interleukin-2
T-Lymphocytes
Messenger RNA
Actins
Ionomycin
Cell Survival
Air
Incubators
Housekeeping
Helium
T-Cell Lymphoma
Phorbol Esters
Tetradecanoylphorbol Acetate
Immunoenzyme Techniques
Culture Media
Hypoxia
Cell Count
Gases
Oxygen
Cell Line

Keywords

  • blood gases
  • cell hypoxia
  • critical illness
  • cytokines
  • interleukin-2
  • ionomycin
  • oxygen
  • phorbol esters
  • RNA, messenger
  • T lymphocytes

ASJC Scopus subject areas

  • Critical Care and Intensive Care Medicine

Cite this

Effects of hypoxia on interleukin-2 mRNA expression by T lymphocytes. / Zuckerberg, A. L.; Goldberg, L. I.; Lederman, Howard M.

In: Critical Care Medicine, Vol. 22, No. 2, 1994, p. 197-203.

Research output: Contribution to journalArticle

Zuckerberg, A. L. ; Goldberg, L. I. ; Lederman, Howard M. / Effects of hypoxia on interleukin-2 mRNA expression by T lymphocytes. In: Critical Care Medicine. 1994 ; Vol. 22, No. 2. pp. 197-203.
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