Quantitative analysis of cytokine mRNA expression in peripheral blood mononuclear cells following treatment with interleukin-2.
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.
Adachi
M
M
Department of Surgery, Medical Institute of Bioregulation, Kyushu University, Beppu, Japan.
Inoue
H
H
Arinaga
S
S
Li
J
J
Ueo
H
H
Mori
M
M
Akiyoshi
T
T
D002277:Carcinoma / Q000097:blood / Q000188:drug therapy; D002448:Cell Adhesion / Q000502:physiology; D016207:Cytokines / Q000096:biosynthesis* / Q000097:blood; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis / Q000097:blood; D007376:Interleukin-2 / Q000494:pharmacology*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000187:drug effects; D009369:Neoplasms / Q000097:blood / Q000188:drug therapy; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000097:blood*; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D016207:Cytokines; D007375:Interleukin-1; D007376:Interleukin-2; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha