Top

Published in:

01-04-2007 | Original Article

Synthesis of glycosylated human tumor necrosis factor α coupled with N-acetylneuraminic acid

Authors: Akiko Hayashi, Taku Chiba, Hidetoshi Hayashi, Satoshi Sasayama, Toshiyuki Ishiguro, Kikuo Onozaki

Published in: Cancer Immunology, Immunotherapy | Issue 4/2007

Abstract

In order to study the effect of glycosylation on its biological activities, and to develop TNFα with less deleterious effects, recombinant human TNFα was chemically coupled with N-acetylneuraminic acid (NeuAc). NeuAc with C9 spacer was coupled to TNFα by acyl azide method. Two glycosylated TNFαs, designated L NeuAc-TNFα and H NeuAc-TNFα, were purified by anion-exchange chromatography. NeuAc coupling to TNFα was confirmed by lectin blotting. Average number of carbohydrate molecules introduced per molecule of L NeuAc-TNFα and H NeuAc-TNFα were estimated to be 1.0 and 1.5, respectively. We examined a variety of TNFα activities in vitro, including antiproliferative or cytotoxic activities to tumor cells, proliferative effect on fibroblast cells, stimulatory effects on IL-6 production by melanoma cells and NF-κB activation in hepatoma cells. L NeuAc-TNFα and H NeuAc-TNFα exhibited reduced activities about 1/3 and 1/10 as compared to native TNFα in all the activities performed in vitro.

Tanaka T, Naruto M, Kawano G (1986) Production of recombinant mouse beta interferon. J Interferon Res 6:429–435PubMed

Tsuda E, Kawanishi G, Ueda M, Masuda S, Sasaki R (1990) The role of carbohydrate in recombinant human erythropoietin. Eur J Biochem 188:405–411PubMedCrossRef

Fischer T, Thoma B, Scheurich P, Pfizenmaier K (1990) Glycosylation of the human interferon-gamma receptor. N-linked carbohydrates contribute to structural heterogeneity and are required for ligand binding. J Biol Chem 265:1710– 1717PubMed

Mancilla J, Ikejima T, Dinarello CA (1992) Glycosylation of the interleukin-1 receptor type I is required for optimal binding of interleukin-1. Lymphokine Cytokine Res 11:197–205PubMed

Ashwell G, Harford J (1982) Carbohydrate-specific receptors of the liver. Annu Rev Biochem 51:531–554PubMedCrossRef

Webb LM, Ehrengruber MU, Clark-Lewis I, Baggiolini M, Rot A (1993) Binding to heparan sulfate or heparin enhances neutrophil responses to interleukin 8. Proc Natl Acad Sci USA. 90:7158–7162PubMedCrossRef

Lopez-Casillas F, Cheifetz S, Doody J, Andres JL, Lane WS, Massague J. (1991) Structure and expression of the membrane proteoglycan betaglycan, a component of the TGF-beta receptor system. Cell 67:785–795PubMedCrossRef

Old J (1985) Tumor necrosis factor (TNF). Science 230:6630–6632CrossRef

Beutler B, Cerami A (1988) Tumor necrosis, cachexia, shock, and inflammation: a common mediator. Ann Rev Biochem 57:505–518PubMedCrossRef

10.

Takei Y, Wada K, Chiba T, Hayashi H, Ishihara H, Onozaki K (1994) Development of glycosylated human interleukin-1 alpha, neoglyco IL-1 alpha, coupled with D-mannose dimer: synthesis and biological activities in vitro. Lymphokine Cytokine Res 13:265–270PubMed

11.

Takei Y, Wada K, Chiba T, Hayashi H, Yamada M, Kuwashima J, Onozaki K (1995) Glycosylated human recombinant interleukin-1 alpha, neo interleukin-1 alpha, with D-mannose dimmer exhibits selective activities in vivo. Lymphokine Cytokine Res 15:713–719

12.

Takei Y, Yang D, Chiba T, Nabeshima S, Naruoka M, Wada K, Onozaki K (1996) D-mannose dimmer introduced human recombinant interleukin-1 alpha, neo IL-1 alpha exhibits altered tissue distribution in mice. J Interferon Cytokine Res 16:333–336PubMed

13.

Chiba T, Nabeshima S, Takei Y, Onozaki K (1998) Development of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, by coupling with D-galactose monosaccharide: synthesis and purification. Glycoconjugate J 15:63–67CrossRef

14.

Nabeshima S, Chiba T, Takei Y, Watanabe S, Okuyama H, Onozaki K (1998) Development of glycosylated human interleukin-1alpha, neoglyco IL-1 alpha, by coupling with D-galactose monosaccharide: biological activities in vitro. Glycoconjugate J 15:69–74CrossRef

15.

Nabeshima S, Chiba T, Takei Y, Ono A, Moriya K, Onozaki K (1998) Development of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with D-galactose monosaccharide: biological activities in vivo. Glycoconjugate J 15:491–498CrossRef

16.

Schauer R (2004) Sialic acids: fascinating sugars in higher animals and man. Zoology (Jena) 107:49–64

17.

Ashwell G, Morell AG (1974) The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv Enzymol Relat Areas Mol Biol 41:99–128PubMedCrossRef

18.

van Rijk, Heinsius HL, van den Hamer CJ (1976) Preparation of 131I-asialo-alpha1-acid glycoprotein. Vox Sang 30:412–419PubMedCrossRef

19.

Crocker PR, Varki A. (2001) Siglecs, sialic acids and innate immunity. Trends Immunol 22:337–42PubMedCrossRef

20.

Crocker PR (2005) Siglecs in innate immunity. Curr Opin Pharmacol 5:431–437PubMedCrossRef

21.

Chiba T, Moriya K, Nabeshima S, Hayashi H, Kobayashi Y, Sasayama S, Onozaki K. (1999) Synthesis of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with N-acetylneuraminic acid. Glycoconj J 16:499–505PubMedCrossRef

22.

Moriya K, Chiba T, Nabeshima S, Hayashi H, Onozaki K (1999) In vitro biological activities of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with N-acetylneuraminic acid. Glycoconj J 16:563–568PubMedCrossRef

23.

Sasayama S, Moriya K, Chiba T, Matsumura T, Hayashi H, Hayashi A, Onozaki K (2000) Glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with N-acetylneuraminic acid exhibits selective activities in vivo and altered tissue distribution. Glycoconj J 17:353–359PubMedCrossRef

24.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227:680–685CrossRef

25.

Matsuda T, Hirano T, Kishimoto T (1988) 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 18:951–956PubMed

26.

Onozaki K, Matsushima K, Aggarwal BB, Oppenheim JJ (1985) Human interleukin 1 is a cytocidal factor for several tumor cell lines. J Immunol 135:3962–3968PubMed

27.

Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63PubMedCrossRef

28.

Hattori T, Hayashi H, Chiba T, Onozaki K (2001) Activation of two distinct anti-proliferative pathways, apoptosis and p38 MAP kinase-dependent cell cycle arrest, by tumor necrosis factor in human melanoma cell line A375. Eur Cytokine Netw 12:244–252PubMed

29.

Sato T., Asamitsu K., Yang JP, Takahashi N, Tetsuka T, Yoneyama A, Kanagawa A., Okamoto T (1998) Inhibition of human immunodeficiency virus type 1 replication by a bioavailable serine/threonine kinase inhibitor, fasudil hydrochloride. AIDS Res Hum Retroviruses 14:293–298PubMedCrossRef

30.

Pennica D, Nedwin GE, Hayflick JS, Seeburg PF, Derynck R, Palladino MA, Kohr WJ, Aggarwal BB, Goeddel DV (1984) Nature 312:724–729PubMedCrossRef

31.

Eck MJ, Sprang SR (1989) Human tumour necrosis factor: precursor structure, expression and homology to lymphotoxin. J Biol Chem 264:17595–17605PubMed

32.

Tartaglia LA, Weber RF, Figari IS, Reynolds C, Palladino MA Jr, Goeddel DV (1991) The two different receptors for tumor necrosis factor mediate distinct cellular responses. Proc Natl Acad Sci USA 88:9292–9296PubMedCrossRef

33.

Lewis M, Tartaglia LA, Lee A, Bennett GL, Rice GC, Wong GH, Chen EV, Goeddel DV (1991) Cloning and expression of cDNAs for two distinct murine tumor necrosis factor receptors demonstrate one receptor is species specific. Proc Natl Acad Sci USA 88:2830–2834PubMedCrossRef

Title: Synthesis of glycosylated human tumor necrosis factor α coupled with N-acetylneuraminic acid
Authors: Akiko Hayashi
Taku Chiba
Hidetoshi Hayashi
Satoshi Sasayama
Toshiyuki Ishiguro
Kikuo Onozaki
Publication date: 01-04-2007
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 4/2007
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-006-0209-8

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 4/2007

R-Ras promotes metastasis of cervical cancer epithelial cells

Vaccination therapy in prostate cancer

International Meeting “Immunotherapy of Cancer: Challenges and Needs”

Decreased response rates by the combination of histamine and IL-2 in melphalan-based isolated limb perfusion

huBC1-IL12, an immunocytokine which targets EDB-containing oncofetal fibronectin in tumors and tumor vasculature, shows potent anti-tumor activity in human tumor models

N-acetylneuraminic acid coupled human recombinant TNFα exhibits enhanced anti-tumor activity against Meth-A fibrosarcoma and reduced toxicity

Keynote webinar | Spotlight on antibody–drug conjugates in cancer