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. 1996 Jan 1;97(1):244–249. doi: 10.1172/JCI118398

Interleukin 6 receptor antibody inhibits muscle atrophy and modulates proteolytic systems in interleukin 6 transgenic mice.

T Tsujinaka 1, J Fujita 1, C Ebisui 1, M Yano 1, E Kominami 1, K Suzuki 1, K Tanaka 1, A Katsume 1, Y Ohsugi 1, H Shiozaki 1, M Monden 1
PMCID: PMC507086  PMID: 8550842

Abstract

The muscles of IL-6 transgenic mice suffer from atrophy. Experiments were carried out on these transgenic mice to elucidate activation of proteolytic systems in the gastrocnemius muscles and blockage of this activation by treatment with the anti-mouse IL-6 receptor (mIL-6R) antibody. Muscle atrophy observed in 16-wk-old transgenic mice was completely blocked by treatment with the mIL-6R antibody. In association with muscle atrophy, enzymatic activities and mRNA levels of cathepsins (B and L) and mRNA levels of ubiquitins (poly- and mono-ubiquitins) increased, whereas the mRNA level of muscle-specific calpain (calpain 3) decreased. All these changes were completely eliminated by treatment with the mIL-6R antibody. This IL-6 receptor antibody could, therefore, be effective against muscle wasting in sepsis and cancer cachexia, where IL-6 plays an important role.

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Selected References

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  1. Bando Y., Kominami E., Katunuma N. Purification and tissue distribution of rat cathepsin L. J Biochem. 1986 Jul;100(1):35–42. doi: 10.1093/oxfordjournals.jbchem.a121703. [DOI] [PubMed] [Google Scholar]
  2. Baracos V. E., DeVivo C., Hoyle D. H., Goldberg A. L. Activation of the ATP-ubiquitin-proteasome pathway in skeletal muscle of cachectic rats bearing a hepatoma. Am J Physiol. 1995 May;268(5 Pt 1):E996–1006. doi: 10.1152/ajpendo.1995.268.5.E996. [DOI] [PubMed] [Google Scholar]
  3. Barrett A. J., Kirschke H. Cathepsin B, Cathepsin H, and cathepsin L. Methods Enzymol. 1981;80(Pt 100):535–561. doi: 10.1016/s0076-6879(81)80043-2. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. Dubiel W., Ferrell K., Pratt G., Rechsteiner M. Subunit 4 of the 26 S protease is a member of a novel eukaryotic ATPase family. J Biol Chem. 1992 Nov 15;267(32):22699–22702. [PubMed] [Google Scholar]
  7. Dubiel W., Ferrell K., Rechsteiner M. Peptide sequencing identifies MSS1, a modulator of HIV Tat-mediated transactivation, as subunit 7 of the 26 S protease. FEBS Lett. 1993 Jun 1;323(3):276–278. doi: 10.1016/0014-5793(93)81356-5. [DOI] [PubMed] [Google Scholar]
  8. Ebisui C., Tsujinaka T., Morimoto T., Kan K., Iijima S., Yano M., Kominami E., Tanaka K., Monden M. Interleukin-6 induces proteolysis by activating intracellular proteases (cathepsins B and L, proteasome) in C2C12 myotubes. Clin Sci (Lond) 1995 Oct;89(4):431–439. doi: 10.1042/cs0890431. [DOI] [PubMed] [Google Scholar]
  9. Fagan J. M., Waxman L., Goldberg A. L. Skeletal muscle and liver contain a soluble ATP + ubiquitin-dependent proteolytic system. Biochem J. 1987 Apr 15;243(2):335–343. doi: 10.1042/bj2430335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fong Y., Moldawer L. L., Marano M., Wei H., Barber A., Manogue K., Tracey K. J., Kuo G., Fischman D. A., Cerami A. Cachectin/TNF or IL-1 alpha induces cachexia with redistribution of body proteins. Am J Physiol. 1989 Mar;256(3 Pt 2):R659–R665. doi: 10.1152/ajpregu.1989.256.3.R659. [DOI] [PubMed] [Google Scholar]
  11. Fujinami K., Tanahashi N., Tanaka K., Ichihara A., Cejka Z., Baumeister W., Miyawaki M., Sato T., Nakagawa H. Purification and characterization of the 26 S proteasome from spinach leaves. J Biol Chem. 1994 Oct 14;269(41):25905–25910. [PubMed] [Google Scholar]
  12. Furuno K., Goodman M. N., Goldberg A. L. Role of different proteolytic systems in the degradation of muscle proteins during denervation atrophy. J Biol Chem. 1990 May 25;265(15):8550–8557. [PubMed] [Google Scholar]
  13. García-Martínez C., Agell N., Llovera M., López-Soriano F. J., Argilés J. M. Tumour necrosis factor-alpha increases the ubiquitinization of rat skeletal muscle proteins. FEBS Lett. 1993 Jun 1;323(3):211–214. doi: 10.1016/0014-5793(93)81341-v. [DOI] [PubMed] [Google Scholar]
  14. García-Martínez C., Llovera M., Agell N., López-Soriano F. J., Argilés J. M. Ubiquitin gene expression in skeletal muscle is increased by tumour necrosis factor-alpha. Biochem Biophys Res Commun. 1994 Jun 15;201(2):682–686. doi: 10.1006/bbrc.1994.1754. [DOI] [PubMed] [Google Scholar]
  15. García-Martínez C., López-Soriano F. J., Argilés J. M. Acute treatment with tumour necrosis factor-alpha induces changes in protein metabolism in rat skeletal muscle. Mol Cell Biochem. 1993 Aug 11;125(1):11–18. doi: 10.1007/BF00926829. [DOI] [PubMed] [Google Scholar]
  16. Goodman M. N. Interleukin-6 induces skeletal muscle protein breakdown in rats. Proc Soc Exp Biol Med. 1994 Feb;205(2):182–185. doi: 10.3181/00379727-205-43695. [DOI] [PubMed] [Google Scholar]
  17. Hall-Angerås M., Hasselgren P. O., Dimlich R. V., Fischer J. E. Myofibrillar proteinase, cathepsin B, and protein breakdown rates in skeletal muscle from septic rats. Metabolism. 1991 Mar;40(3):302–306. doi: 10.1016/0026-0495(91)90114-c. [DOI] [PubMed] [Google Scholar]
  18. Hershko A., Ciechanover A. The ubiquitin system for protein degradation. Annu Rev Biochem. 1992;61:761–807. doi: 10.1146/annurev.bi.61.070192.003553. [DOI] [PubMed] [Google Scholar]
  19. Hirata Y., Taga T., Hibi M., Nakano N., Hirano T., Kishimoto T. Characterization of IL-6 receptor expression by monoclonal and polyclonal antibodies. J Immunol. 1989 Nov 1;143(9):2900–2906. [PubMed] [Google Scholar]
  20. Ishidoh K., Towatari T., Imajoh S., Kawasaki H., Kominami E., Katunuma N., Suzuki K. Molecular cloning and sequencing of cDNA for rat cathepsin L. FEBS Lett. 1987 Oct 19;223(1):69–73. doi: 10.1016/0014-5793(87)80511-2. [DOI] [PubMed] [Google Scholar]
  21. Johnson P. Calpains (intracellular calcium-activated cysteine proteinases): structure-activity relationships and involvement in normal and abnormal cellular metabolism. Int J Biochem. 1990;22(8):811–822. doi: 10.1016/0020-711x(90)90284-a. [DOI] [PubMed] [Google Scholar]
  22. Kanayama H., Tanaka K., Aki M., Kagawa S., Miyaji H., Satoh M., Okada F., Sato S., Shimbara N., Ichihara A. Changes in expressions of proteasome and ubiquitin genes in human renal cancer cells. Cancer Res. 1991 Dec 15;51(24):6677–6685. [PubMed] [Google Scholar]
  23. Kettelhut I. C., Goldberg A. L. Tumor necrosis factor can induce fever in rats without activating protein breakdown in muscle or lipolysis in adipose tissue. J Clin Invest. 1988 May;81(5):1384–1389. doi: 10.1172/JCI113467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kominami E., Kunio I., Katunuma N. Activation of the intramyofibral autophagic-lysosomal system in muscular dystrophy. Am J Pathol. 1987 Jun;127(3):461–466. [PMC free article] [PubMed] [Google Scholar]
  25. Kominami E., Tsukahara T., Bando Y., Katunuma N. Distribution of cathepsins B and H in rat tissues and peripheral blood cells. J Biochem. 1985 Jul;98(1):87–93. doi: 10.1093/oxfordjournals.jbchem.a135277. [DOI] [PubMed] [Google Scholar]
  26. Libert C., Brouckaert P., Shaw A., Fiers W. Induction of interleukin 6 by human and murine recombinant interleukin 1 in mice. Eur J Immunol. 1990 Mar;20(3):691–694. doi: 10.1002/eji.1830200333. [DOI] [PubMed] [Google Scholar]
  27. Lowell B. B., Ruderman N. B., Goodman M. N. Evidence that lysosomes are not involved in the degradation of myofibrillar proteins in rat skeletal muscle. Biochem J. 1986 Feb 15;234(1):237–240. doi: 10.1042/bj2340237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lundholm K., Bylund A. C., Holm J., Scherstén T. Skeletal muscle metabolism in patients with malignant tumor. Eur J Cancer. 1976 Jun;12(6):465–473. doi: 10.1016/0014-2964(76)90036-0. [DOI] [PubMed] [Google Scholar]
  29. Matsuda T., Hirano T., Kishimoto T. Establishment of an interleukin 6 (IL 6)/B cell stimulatory factor 2-dependent cell line and preparation of anti-IL 6 monoclonal antibodies. Eur J Immunol. 1988 Jun;18(6):951–956. doi: 10.1002/eji.1830180618. [DOI] [PubMed] [Google Scholar]
  30. Medina R., Wing S. S., Goldberg A. L. Increase in levels of polyubiquitin and proteasome mRNA in skeletal muscle during starvation and denervation atrophy. Biochem J. 1995 May 1;307(Pt 3):631–637. doi: 10.1042/bj3070631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mitch W. E., Medina R., Grieber S., May R. C., England B. K., Price S. R., Bailey J. L., Goldberg A. L. Metabolic acidosis stimulates muscle protein degradation by activating the adenosine triphosphate-dependent pathway involving ubiquitin and proteasomes. J Clin Invest. 1994 May;93(5):2127–2133. doi: 10.1172/JCI117208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Moldawer L. L., Svaninger G., Gelin J., Lundholm K. G. Interleukin 1 and tumor necrosis factor do not regulate protein balance in skeletal muscle. Am J Physiol. 1987 Dec;253(6 Pt 1):C766–C773. doi: 10.1152/ajpcell.1987.253.6.C766. [DOI] [PubMed] [Google Scholar]
  33. Rechsteiner M. Natural substrates of the ubiquitin proteolytic pathway. Cell. 1991 Aug 23;66(4):615–618. doi: 10.1016/0092-8674(91)90104-7. [DOI] [PubMed] [Google Scholar]
  34. Richard I., Broux O., Allamand V., Fougerousse F., Chiannilkulchai N., Bourg N., Brenguier L., Devaud C., Pasturaud P., Roudaut C. Mutations in the proteolytic enzyme calpain 3 cause limb-girdle muscular dystrophy type 2A. Cell. 1995 Apr 7;81(1):27–40. doi: 10.1016/0092-8674(95)90368-2. [DOI] [PubMed] [Google Scholar]
  35. Sato K., Tsuchiya M., Saldanha J., Koishihara Y., Ohsugi Y., Kishimoto T., Bendig M. M. Reshaping a human antibody to inhibit the interleukin 6-dependent tumor cell growth. Cancer Res. 1993 Feb 15;53(4):851–856. [PubMed] [Google Scholar]
  36. Sehgal P. B. Interleukin 6 in infection and cancer. Proc Soc Exp Biol Med. 1990 Nov;195(2):183–191. doi: 10.3181/00379727-195-43129d. [DOI] [PubMed] [Google Scholar]
  37. Sorimachi H., Imajoh-Ohmi S., Emori Y., Kawasaki H., Ohno S., Minami Y., Suzuki K. Molecular cloning of a novel mammalian calcium-dependent protease distinct from both m- and mu-types. Specific expression of the mRNA in skeletal muscle. J Biol Chem. 1989 Nov 25;264(33):20106–20111. [PubMed] [Google Scholar]
  38. Sorimachi H., Saido T. C., Suzuki K. New era of calpain research. Discovery of tissue-specific calpains. FEBS Lett. 1994 Apr 18;343(1):1–5. doi: 10.1016/0014-5793(94)80595-4. [DOI] [PubMed] [Google Scholar]
  39. Strassmann G., Fong M., Freter C. E., Windsor S., D'Alessandro F., Nordan R. P. Suramin interferes with interleukin-6 receptor binding in vitro and inhibits colon-26-mediated experimental cancer cachexia in vivo. J Clin Invest. 1993 Nov;92(5):2152–2159. doi: 10.1172/JCI116816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Strassmann G., Fong M., Kenney J. S., Jacob C. O. Evidence for the involvement of interleukin 6 in experimental cancer cachexia. J Clin Invest. 1992 May;89(5):1681–1684. doi: 10.1172/JCI115767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Suematsu S., Matsuda T., Aozasa K., Akira S., Nakano N., Ohno S., Miyazaki J., Yamamura K., Hirano T., Kishimoto T. IgG1 plasmacytosis in interleukin 6 transgenic mice. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7547–7551. doi: 10.1073/pnas.86.19.7547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Suematsu S., Matsusaka T., Matsuda T., Ohno S., Miyazaki J., Yamamura K., Hirano T., Kishimoto T. Generation of plasmacytomas with the chromosomal translocation t(12;15) in interleukin 6 transgenic mice. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):232–235. doi: 10.1073/pnas.89.1.232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Suzuki H., Yasukawa K., Saito T., Goitsuka R., Hasegawa A., Ohsugi Y., Taga T., Kishimoto T. Anti-human interleukin-6 receptor antibody inhibits human myeloma growth in vivo. Eur J Immunol. 1992 Aug;22(8):1989–1993. doi: 10.1002/eji.1830220806. [DOI] [PubMed] [Google Scholar]
  44. Tamura T., Lee D. H., Osaka F., Fujiwara T., Shin S., Chung C. H., Tanaka K., Ichihara A. Molecular cloning and sequence analysis of cDNAs for five major subunits of human proteasomes (multi-catalytic proteinase complexes). Biochim Biophys Acta. 1991 May 2;1089(1):95–102. doi: 10.1016/0167-4781(91)90090-9. [DOI] [PubMed] [Google Scholar]
  45. Tamura T., Udagawa N., Takahashi N., Miyaura C., Tanaka S., Yamada Y., Koishihara Y., Ohsugi Y., Kumaki K., Taga T. Soluble interleukin-6 receptor triggers osteoclast formation by interleukin 6. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11924–11928. doi: 10.1073/pnas.90.24.11924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Temparis S., Asensi M., Taillandier D., Aurousseau E., Larbaud D., Obled A., Béchet D., Ferrara M., Estrela J. M., Attaix D. Increased ATP-ubiquitin-dependent proteolysis in skeletal muscles of tumor-bearing rats. Cancer Res. 1994 Nov 1;54(21):5568–5573. [PubMed] [Google Scholar]
  47. Tsujinaka T., Ebisui C., Fujita J., Kishibuchi M., Morimoto T., Ogawa A., Katsume A., Ohsugi Y., Kominami E., Monden M. Muscle undergoes atrophy in association with increase of lysosomal cathepsin activity in interleukin-6 transgenic mouse. Biochem Biophys Res Commun. 1995 Feb 6;207(1):168–174. doi: 10.1006/bbrc.1995.1168. [DOI] [PubMed] [Google Scholar]
  48. Tågerud S., Libelius R. Receptor-mediated uptake of horseradish peroxidase in innervated and denervated skeletal muscle. Exp Cell Res. 1985 Sep;160(1):95–105. doi: 10.1016/0014-4827(85)90239-3. [DOI] [PubMed] [Google Scholar]
  49. Warren R. S., Starnes H. F., Jr, Gabrilove J. L., Oettgen H. F., Brennan M. F. The acute metabolic effects of tumor necrosis factor administration in humans. Arch Surg. 1987 Dec;122(12):1396–1400. doi: 10.1001/archsurg.1987.01400240042007. [DOI] [PubMed] [Google Scholar]
  50. Wing S. S., Goldberg A. L. Glucocorticoids activate the ATP-ubiquitin-dependent proteolytic system in skeletal muscle during fasting. Am J Physiol. 1993 Apr;264(4 Pt 1):E668–E676. doi: 10.1152/ajpendo.1993.264.4.E668. [DOI] [PubMed] [Google Scholar]
  51. Yamamura K., Kikutani H., Takahashi N., Taga T., Akira S., Kawai K., Fukuchi K., Kumahara Y., Honjo T., Kishimoto T. Introduction of human gamma 1 immunoglobulin genes into fertilized mouse eggs. J Biochem. 1984 Aug;96(2):357–363. doi: 10.1093/oxfordjournals.jbchem.a134845. [DOI] [PubMed] [Google Scholar]

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