Quantitative analysis of cytokine mRNA expression in peripheral blood mononuclear cells following treatment with interleukin-2

Cancer Immunol Immunother. 1997 Aug;44(6):329-34. doi: 10.1007/s002620050390.

Abstract

After activation with interleukin-2 (IL-2), peripheral blood mononuclear cells (PBMC) have been reported to induce the expression of mRNA coding various cytokines, including interleukin(IL)-1alpha, -1beta and tumor necrosis factor alpha (TNF alpha). We examined the cytokine mRNA expression of PBMC following treatment with IL-2 in vitro and in vivo by a quantitative method using the reverse transcription/polymerase chain reaction (RT-PCR). After stimulating PBMC with IL-2 in vitro, peak levels of IL-1alpha mRNA were reached between 3 h and 12 h, and thereafter declined. The IL-1beta expression increased, with levels peaking at 1-6 h and, had decreased by 96 h. The expression of TNF alpha was elevated both 1-3 h and 24-48 h after stimulation. The peak levels of IL-1alpha and -1beta mRNA and the early elevation of TNF alpha mRNA mainly accounted for the cytokine mRNA expression in adherent cells; however, the late induction of TNF alpha mRNA was observed in nonadherent cells. In patients with advanced carcinoma, the IL-1alpha and -1beta mRNA expression were elevated after IL-2 treatment for 5 consecutive days, while the expression of TNF alpha mRNA also increased. These results indicate that the quantitative RT-PCR method appears to be useful for analyzing the cytokine mRNA expression of PBMC after treatment with IL-2.

MeSH terms

  • Carcinoma / blood
  • Carcinoma / drug therapy
  • Cell Adhesion / physiology
  • Cytokines / biosynthesis*
  • Cytokines / blood
  • Humans
  • Interleukin-1 / biosynthesis
  • Interleukin-1 / blood
  • Interleukin-2 / pharmacology*
  • Kinetics
  • Leukocytes, Mononuclear / drug effects*
  • Leukocytes, Mononuclear / metabolism*
  • Lymphocyte Activation / drug effects
  • Neoplasms / blood
  • Neoplasms / drug therapy
  • Polymerase Chain Reaction
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / blood*
  • Transcription, Genetic
  • Tumor Necrosis Factor-alpha / biosynthesis

Substances

  • Cytokines
  • Interleukin-1
  • Interleukin-2
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha