Specific inhibition of interleukin 1 beta gene expression by an antisense oligonucleotide: obligatory role of interleukin 1 in the generation of lymphokine-activated killer cells.
The purpose of the studies reported here is to determine whether interleukin 1 (IL-1) plays an important role in the regulation of lymphokine-activated killer (LAK) cell induction. The addition of exogenous IL-1 to peripheral blood lymphocyte culture containing suboptimal concentrations of interleukin 2 (IL-2) resulted in induction of cytoplasmic pore-forming protein expression. Polymerase chain reaction results revealed that the mRNAs of both IL-1 alpha and IL-1 beta were induced within 6 h when cultured in IL-2 alone or in a combination of IL-2 and IL-1; however, tumor necrosis factor alpha and beta mRNAs were expressed earlier in peripheral blood lymphocytes stimulated with the combination of IL-1 and IL-2. Furthermore, we have examined the functional role of endogenous IL-1 in LAK activity. The lytic potential was significantly inhibited by an IL-1 receptor antagonist, which could block IL-1-mediated effects, or by specific neutralizing antibodies for IL-1, suggesting that the extracellular autocrine/paracrine pathway of IL-1 is involved in LAK activation. However, a synthetic IL-1 beta antisense oligonucleotide, which could specifically inhibit intracellular IL-1 beta protein expression as detected by Western blot, was more effective in reducing LAK killing, but it could not suppress the cytotoxicity generated by exogenous IL-1 plus IL-2. These findings clearly indicate the existence of an intracellular IL-1 autocrine circuit. Taken together, our results strongly indicate that IL-1 should be considered an obligatory factor in the regulation of IL-2-mediated lymphocyte functions.
Fujiwara
T
T
Department of Tumor Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Grimm
E A
EA
D001483:Base Sequence; D016207:Cytokines / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000235:genetics / Q000502:physiology*; D007376:Interleukin-2 / Q000494:pharmacology; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology*; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016376:Oligonucleotides, Antisense; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D011506:Proteins / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D012795:Sialoglycoproteins
D016207:Cytokines; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D052899:Pore Forming Cytotoxic Proteins; D011506:Proteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D054353:Perforin