T-cell interleukin 1 receptor cDNA expressed in Chinese hamster ovary cells regulates functional responses to interleukin 1.
We have cloned a cDNA encoding a receptor identical to the native Mr 80,000 glycoprotein that binds interleukin (IL) 1 alpha and -beta in murine T cells. Chinese hamster ovary (CHO) cells transfected with this T-cell IL-1 receptor (IL-1R) [CHO(IL-1R)] cDNA express approximately 100,000 IL-1Rs per cell, compared to the less than 100 receptors present on control CHO cells. For two functional responses to IL-1, prostaglandin synthesis and cytokine secretion, CHO(IL-1R) cells were 1000 times more sensitive to IL-1 alpha than were control CHO cells. Northern blot analysis and antibody precipitation demonstrated that one of the cytokines induced was granulocyte colony-stimulating factor and that mRNA levels for this cytokine were increased in CHO(IL-1R) cells by IL-1 alpha concentrations that had no effect on control cells. To establish the role of the recombinant receptors in signal transduction, an IL-1R cDNA modified by deletion of the predicted cytoplasmic domain was expressed in the CHO cell line termed CHO(IL-1R delta CT). CHO(IL-1R delta CT) cells expressed approximately 100,000 high-affinity IL-1 binding sites per cell, but these cells were less sensitive than control lines to IL-1, as measured by prostaglandin and cytokine release. These results show that the IL-1R cDNA encodes the entire functional receptor and that the cytoplasmic domain is required for signal transduction but not ligand binding.
Curtis
B M
BM
Department of Protein Chemistry, Immunex Corp., Seattle, WA 98101.
Gallis
B
B
Overell
R W
RW
McMahan
C J
CJ
DeRoos
P
P
Ireland
R
R
Eisenman
J
J
Dower
S K
SK
Sims
J E
JE
D000818:Animals; D002460:Cell Line; D003115:Colony-Stimulating Factors / Q000096:biosynthesis / Q000235:genetics; D006224:Cricetinae; D004247:DNA / Q000235:genetics*; D015232:Dinoprostone / Q000096:biosynthesis; D005786:Gene Expression Regulation; D016179:Granulocyte Colony-Stimulating Factor; D007375:Interleukin-1 / Q000502:physiology*; D007700:Kinetics; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D011971:Receptors, Immunologic / Q000235:genetics*; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
D003115:Colony-Stimulating Factors; D007375:Interleukin-1; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D004247:DNA; D015232:Dinoprostone