Sequence, expression and function of an mRNA encoding a soluble form of the human interleukin-6 receptor (sIL-6R)

J. A. Lust, D. F. Jelinek, K. A. Donovan, L. A. Frederick, B. K. Huntley, J. K. Braaten, Nita Jane Maihle

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Soluble receptors have been shown to be potent immunomodulators of their respective ligands. Since IL-6 is a central growth factor for myeloma cells, as sIL-6R may modulate IL-6 activity. We have previously reported a novel IL-6R mRNA from myeloma cells that exhibits a 94-nt deletion of the entire transmembrane domain from codons 356 (G-TG) to 387 (AG-G). The transmembrane domain deletion results in a shift in the translational reading frame with the insertion of 10 new amino acids followed by a stop codon. Sequence analysis shows the ligand-binding domain of the sIL-6R to be identical to that of the membrane-bound IL-6R up to the transmembrane domain deletion. The sIL-6R cDNA was expressed in QT-6 fibroblasts and PA-1 ovarian cells using the expression vector pCDM8. Supernates were immunoprecipitated with anti-IL-6R antibody and cells transfected with the sIL-6R cDNA produced a single band with a molecular weight of 50-55 kDa. This molecular weight corresponds to the size of the sIL-6R protein observed in normal human urine. Supernates were collected from mock or sIL-6R transfected PA-1 cells after 48 hours and assayed for their ability to stimulate or suppress the growth of an IL-6 dependent cell line, ANBL-6. Soluble IL-6R alone had no effect on the growth of the ANBL-6 cells. However, the growth of ANBL-6 cells by sIL-6R was potentiated in the presence of IL-6 and could be blocked by anti-IL-6 antibody. The above results suggest that, in the presence of IL-6, sIL-6R associates with gp130 leading to signal transduction and cell growth.

Original languageEnglish (US)
Pages (from-to)199-206
Number of pages8
JournalCurrent Topics in Microbiology and Immunology
Volume194
StatePublished - Jan 1 1994
Externally publishedYes

Fingerprint

Interleukin-6 Receptors
Interleukin-6
Messenger RNA
Growth
Complementary DNA
Molecular Weight
Ligands
Reading Frames
Antibodies
Terminator Codon
Immunologic Factors
Codon
Sequence Analysis
Signal Transduction
Intercellular Signaling Peptides and Proteins
Fibroblasts
Urine
Amino Acids
Cell Line
Membranes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Microbiology
  • Immunology
  • Microbiology (medical)

Cite this

Lust, J. A., Jelinek, D. F., Donovan, K. A., Frederick, L. A., Huntley, B. K., Braaten, J. K., & Maihle, N. J. (1994). Sequence, expression and function of an mRNA encoding a soluble form of the human interleukin-6 receptor (sIL-6R). Current Topics in Microbiology and Immunology, 194, 199-206.

Sequence, expression and function of an mRNA encoding a soluble form of the human interleukin-6 receptor (sIL-6R). / Lust, J. A.; Jelinek, D. F.; Donovan, K. A.; Frederick, L. A.; Huntley, B. K.; Braaten, J. K.; Maihle, Nita Jane.

In: Current Topics in Microbiology and Immunology, Vol. 194, 01.01.1994, p. 199-206.

Research output: Contribution to journalArticle

Lust, JA, Jelinek, DF, Donovan, KA, Frederick, LA, Huntley, BK, Braaten, JK & Maihle, NJ 1994, 'Sequence, expression and function of an mRNA encoding a soluble form of the human interleukin-6 receptor (sIL-6R)', Current Topics in Microbiology and Immunology, vol. 194, pp. 199-206.
Lust, J. A. ; Jelinek, D. F. ; Donovan, K. A. ; Frederick, L. A. ; Huntley, B. K. ; Braaten, J. K. ; Maihle, Nita Jane. / Sequence, expression and function of an mRNA encoding a soluble form of the human interleukin-6 receptor (sIL-6R). In: Current Topics in Microbiology and Immunology. 1994 ; Vol. 194. pp. 199-206.
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abstract = "Soluble receptors have been shown to be potent immunomodulators of their respective ligands. Since IL-6 is a central growth factor for myeloma cells, as sIL-6R may modulate IL-6 activity. We have previously reported a novel IL-6R mRNA from myeloma cells that exhibits a 94-nt deletion of the entire transmembrane domain from codons 356 (G-TG) to 387 (AG-G). The transmembrane domain deletion results in a shift in the translational reading frame with the insertion of 10 new amino acids followed by a stop codon. Sequence analysis shows the ligand-binding domain of the sIL-6R to be identical to that of the membrane-bound IL-6R up to the transmembrane domain deletion. The sIL-6R cDNA was expressed in QT-6 fibroblasts and PA-1 ovarian cells using the expression vector pCDM8. Supernates were immunoprecipitated with anti-IL-6R antibody and cells transfected with the sIL-6R cDNA produced a single band with a molecular weight of 50-55 kDa. This molecular weight corresponds to the size of the sIL-6R protein observed in normal human urine. Supernates were collected from mock or sIL-6R transfected PA-1 cells after 48 hours and assayed for their ability to stimulate or suppress the growth of an IL-6 dependent cell line, ANBL-6. Soluble IL-6R alone had no effect on the growth of the ANBL-6 cells. However, the growth of ANBL-6 cells by sIL-6R was potentiated in the presence of IL-6 and could be blocked by anti-IL-6 antibody. The above results suggest that, in the presence of IL-6, sIL-6R associates with gp130 leading to signal transduction and cell growth.",
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