The relative radioresistance of interleukin-2 production by human peripheral blood lymphocytes: consequences for the development of a new limiting dilution assay for the enumeration of helper T lymphocyte precursor frequencies

J Immunol Methods. 1994 Mar 10;169(2):221-30. doi: 10.1016/0022-1759(94)90266-6.

Abstract

We describe a limiting dilution assay for the enumeration of alloreactive helper T lymphocyte precursor frequencies in human peripheral blood. The proliferation rate of the murine indicator cell line, cytotoxic T lymphoblastic line 2 (CTLL-2) induced by interleukin-2 (IL-2) culture supernatants was determined by staining with the fluorescent DNA dye propidium-iodide. Lymphocytes from healthy individuals as well as from patients with end stage kidney disease and no previous allosensitization exhibited a relative radioresistance of their IL-2 production up to gamma irradiation doses of 40-60 Gy. This differs from previous findings in the literature, showing a total inhibition of the IL-2 production in unsensitized individuals using a gamma irradiation dose of 20 Gy. The consequences of this relative radioresistance are that for a reliable stimulator cell inactivation in assays for the enumeration of helper T lymphocyte precursors gamma irradiation doses of at least 50 (-60) Gy are needed. Increasing the gamma irradiation dose for the inactivation of the stimulator cells can result in a decrease of the antigen presenting capacity of these cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation / immunology
  • Cell Division
  • Cell Line
  • Cell Survival / radiation effects
  • Cells, Cultured
  • DNA / biosynthesis
  • Fetal Blood / immunology
  • HLA-DR Antigens / immunology
  • Humans
  • Immunologic Techniques*
  • Interleukin-2 / biosynthesis*
  • Kidney Failure, Chronic / immunology
  • Kidney Failure, Chronic / pathology
  • Leukocyte Count
  • Mice
  • Radiation Tolerance*
  • Stem Cells / immunology
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / radiation effects
  • T-Lymphocytes, Helper-Inducer / immunology
  • T-Lymphocytes, Helper-Inducer / radiation effects*

Substances

  • HLA-DR Antigens
  • Interleukin-2
  • DNA