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. 1998 Nov;95(3):360–369. doi: 10.1046/j.1365-2567.1998.00622.x

High-level production of alternatively spliced soluble interleukin-6 receptor in serum of patients with adult T-cell leukaemia/HTLV-I-associated myelopathy.

S Horiuchi 1, W Ampofo 1, Y Koyanagi 1, A Yamashita 1, M Waki 1, A Matsumoto 1, M Yamamoto 1, N Yamamoto 1
PMCID: PMC1364401  PMID: 9824498

Abstract

We have previously shown, using human T-cell lymphocytotrophic virus-I (HTLV-I)-infected cell lines, that soluble interleukin-6 receptor (sIL-6R) is generated through an alternative splicing mechanism. In this study, we examined human sera for the presence of alternatively spliced soluble IL-6R (AS-sIL-6R). We produced a monoclonal antibody (mAb) recognizing the unique sequence of AS-sIL-6R peptide, generated by an altered reading frame. We also made recombinant AS-sIL-6R protein in Spodoptera frugiperda-9 (Sf-9) cells carrying baculovirus, which encoded altered sIL-6R or conventional IL-6R cDNA. mAbs specifically recognized AS-sIL-6R, but not conventional IL-6R, as demonstrated by Western blot analyses, fluorescence-activated cell sorter, immunofluorescence analyses and enzyme-linked immunosorbent assay (ELISA). We adapted an ELISA system and used it for detection of altered sIL-6R in sera from 23 healthy persons, 12 patients with adult T-cell leukaemia (ATL) and 13 patients with HTLV-I-associated myelopathy (HAM). Serum levels of AS-sIL-6R were 6.4 or 6.1 times greater in ATL (28.7+/-20.4 ng/ml, P<0.0001) and in HAM patients (27.5+/-12.1 ng/ml, P<0.0001) than in healthy individuals (4.5+/-2.1 ng/ml). High levels of AS-sIL-6R were also observed in plasma from rheumatoid arthritis patients and in persons with elevated levels of alanine aminotransferase (ALT), antinuclear antibody (ANA), or alpha-fetoprotein (AFP). However, in human immunodeficiency virus-1 (HIV-1), hepatitis B virus (HBV) or hepatitis C virus (HCV)-infected individuals, AS-sIL-6R levels were not elevated. In this study, we confirmed that AS-sIL-6R is indeed present in human sera. These observations suggest that alternative splicing of IL-6R mRNA is of consequence in ATL, HAM and in some autoimmune diseases. The HTLV-I-infected T cells appeared to play an important role in AS-sIL-6R production.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. De Benedetti F., Massa M., Pignatti P., Albani S., Novick D., Martini A. Serum soluble interleukin 6 (IL-6) receptor and IL-6/soluble IL-6 receptor complex in systemic juvenile rheumatoid arthritis. J Clin Invest. 1994 May;93(5):2114–2119. doi: 10.1172/JCI117206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gabay C., Silacci P., Genin B., Mentha G., Le Coultre C., Guerne P. A. Soluble interleukin-6 receptor strongly increases the production of acute-phase protein by hepatoma cells but exerts minimal changes on human primary hepatocytes. Eur J Immunol. 1995 Aug;25(8):2378–2383. doi: 10.1002/eji.1830250838. [DOI] [PubMed] [Google Scholar]
  3. Gaillard J. P., Bataille R., Brailly H., Zuber C., Yasukawa K., Attal M., Maruo N., Taga T., Kishimoto T., Klein B. Increased and highly stable levels of functional soluble interleukin-6 receptor in sera of patients with monoclonal gammopathy. Eur J Immunol. 1993 Apr;23(4):820–824. doi: 10.1002/eji.1830230408. [DOI] [PubMed] [Google Scholar]
  4. Gauldie J., Richards C., Harnish D., Lansdorp P., Baumann H. Interferon beta 2/B-cell stimulatory factor type 2 shares identity with monocyte-derived hepatocyte-stimulating factor and regulates the major acute phase protein response in liver cells. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7251–7255. doi: 10.1073/pnas.84.20.7251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hirota H., Yoshida K., Kishimoto T., Taga T. Continuous activation of gp130, a signal-transducing receptor component for interleukin 6-related cytokines, causes myocardial hypertrophy in mice. Proc Natl Acad Sci U S A. 1995 May 23;92(11):4862–4866. doi: 10.1073/pnas.92.11.4862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Holmdahl R., Moran T., Andersson M. A rapid and efficient immunization protocol for production of monoclonal antibodies reactive with autoantigens. J Immunol Methods. 1985 Nov 7;83(2):379–384. doi: 10.1016/0022-1759(85)90260-1. [DOI] [PubMed] [Google Scholar]
  7. Honda M., Yamamoto S., Cheng M., Yasukawa K., Suzuki H., Saito T., Osugi Y., Tokunaga T., Kishimoto T. Human soluble IL-6 receptor: its detection and enhanced release by HIV infection. J Immunol. 1992 Apr 1;148(7):2175–2180. [PubMed] [Google Scholar]
  8. Horiuchi S., Koyanagi Y., Tanaka Y., Waki M., Matsumoto A., Zhou Y. W., Yamamoto M., Yamamoto N. Altered interleukin-2 receptor alpha-chain is expressed in human T-cell leukaemia virus type-I-infected T-cell lines and human peripheral blood mononuclear cells of adult T-cell leukaemia patients through an alternative splicing mechanism. Immunology. 1997 May;91(1):28–34. doi: 10.1046/j.1365-2567.1997.00236.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Horiuchi S., Koyanagi Y., Zhou Y., Miyamoto H., Tanaka Y., Waki M., Matsumoto A., Yamamoto M., Yamamoto N. Soluble interleukin-6 receptors released from T cell or granulocyte/macrophage cell lines and human peripheral blood mononuclear cells are generated through an alternative splicing mechanism. Eur J Immunol. 1994 Aug;24(8):1945–1948. doi: 10.1002/eji.1830240837. [DOI] [PubMed] [Google Scholar]
  10. Hoyland J. A., Freemont A. J., Sharpe P. T. Interleukin-6, IL-6 receptor, and IL-6 nuclear factor gene expression in Paget's disease. J Bone Miner Res. 1994 Jan;9(1):75–80. doi: 10.1002/jbmr.5650090111. [DOI] [PubMed] [Google Scholar]
  11. Ikebuchi K., Wong G. G., Clark S. C., Ihle J. N., Hirai Y., Ogawa M. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9035–9039. doi: 10.1073/pnas.84.24.9035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inoue J., Seiki M., Taniguchi T., Tsuru S., Yoshida M. Induction of interleukin 2 receptor gene expression by p40x encoded by human T-cell leukemia virus type 1. EMBO J. 1986 Nov;5(11):2883–2888. doi: 10.1002/j.1460-2075.1986.tb04583.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lust J. A., Donovan K. A., Kline M. P., Greipp P. R., Kyle R. A., Maihle N. J. Isolation of an mRNA encoding a soluble form of the human interleukin-6 receptor. Cytokine. 1992 Mar;4(2):96–100. doi: 10.1016/1043-4666(92)90043-q. [DOI] [PubMed] [Google Scholar]
  14. Maniatis T. Mechanisms of alternative pre-mRNA splicing. Science. 1991 Jan 4;251(4989):33–34. doi: 10.1126/science.1824726. [DOI] [PubMed] [Google Scholar]
  15. Maruyama M., Shibuya H., Harada H., Hatakeyama M., Seiki M., Fujita T., Inoue J., Yoshida M., Taniguchi T. Evidence for aberrant activation of the interleukin-2 autocrine loop by HTLV-1-encoded p40x and T3/Ti complex triggering. Cell. 1987 Jan 30;48(2):343–350. doi: 10.1016/0092-8674(87)90437-5. [DOI] [PubMed] [Google Scholar]
  16. Mattox W., Ryner L., Baker B. S. Autoregulation and multifunctionality among trans-acting factors that regulate alternative pre-mRNA processing. J Biol Chem. 1992 Sep 25;267(27):19023–19026. [PubMed] [Google Scholar]
  17. Mihara M., Moriya Y., Kishimoto T., Ohsugi Y. Interleukin-6 (IL-6) induces the proliferation of synovial fibroblastic cells in the presence of soluble IL-6 receptor. Br J Rheumatol. 1995 Apr;34(4):321–325. doi: 10.1093/rheumatology/34.4.321. [DOI] [PubMed] [Google Scholar]
  18. Murakami-Mori K., Taga T., Kishimoto T., Nakamura S. The soluble form of the IL-6 receptor (sIL-6R alpha) is a potent growth factor for AIDS-associated Kaposi's sarcoma (KS) cells; the soluble form of gp130 is antagonistic for sIL-6R alpha-induced AIDS-KS cell growth. Int Immunol. 1996 Apr;8(4):595–602. doi: 10.1093/intimm/8.4.595. [DOI] [PubMed] [Google Scholar]
  19. Müllberg J., Oberthür W., Lottspeich F., Mehl E., Dittrich E., Graeve L., Heinrich P. C., Rose-John S. The soluble human IL-6 receptor. Mutational characterization of the proteolytic cleavage site. J Immunol. 1994 May 15;152(10):4958–4968. [PubMed] [Google Scholar]
  20. Ohtani K., Ninomiya H., Hasegawa Y., Kobayashi T., Kojima H., Nagasawa T., Abe T. Clinical significance of elevated soluble interleukin-6 receptor levels in the sera of patients with plasma cell dyscrasias. Br J Haematol. 1995 Sep;91(1):116–120. doi: 10.1111/j.1365-2141.1995.tb05255.x. [DOI] [PubMed] [Google Scholar]
  21. Paonessa G., Graziani R., De Serio A., Savino R., Ciapponi L., Lahm A., Salvati A. L., Toniatti C., Ciliberto G. Two distinct and independent sites on IL-6 trigger gp 130 dimer formation and signalling. EMBO J. 1995 May 1;14(9):1942–1951. doi: 10.1002/j.1460-2075.1995.tb07186.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rose-John S., Heinrich P. C. Soluble receptors for cytokines and growth factors: generation and biological function. Biochem J. 1994 Jun 1;300(Pt 2):281–290. doi: 10.1042/bj3000281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Taga T., Hibi M., Hirata Y., Yamasaki K., Yasukawa K., Matsuda T., Hirano T., Kishimoto T. Interleukin-6 triggers the association of its receptor with a possible signal transducer, gp130. Cell. 1989 Aug 11;58(3):573–581. doi: 10.1016/0092-8674(89)90438-8. [DOI] [PubMed] [Google Scholar]
  24. Udagawa N., Takahashi N., Katagiri T., Tamura T., Wada S., Findlay D. M., Martin T. J., Hirota H., Taga T., Kishimoto T. Interleukin (IL)-6 induction of osteoclast differentiation depends on IL-6 receptors expressed on osteoblastic cells but not on osteoclast progenitors. J Exp Med. 1995 Nov 1;182(5):1461–1468. doi: 10.1084/jem.182.5.1461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yamasaki K., Taga T., Hirata Y., Yawata H., Kawanishi Y., Seed B., Taniguchi T., Hirano T., Kishimoto T. Cloning and expression of the human interleukin-6 (BSF-2/IFN beta 2) receptor. Science. 1988 Aug 12;241(4867):825–828. doi: 10.1126/science.3136546. [DOI] [PubMed] [Google Scholar]

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