Skip to main content
SpringerLink
Log in

Alterations of glycan branching and differential expression of sialic acid on alpha fetoprotein among hepatitis patients

  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

The level of serum glycoproteins and their glycosylation pattern change in liver diseases including hepatocellular carcinoma (HCC). Some of them, especially alpha fetoprotein (AFP), serve as useful biomarkers for HCC. The present investigation showed high level of AFP in hepatitis B cirrhosis (HBV-LC) and hepatitis C cirrhosis (HCV-LC) patients. However, increase of AFP level was not significantly high in chronic hepatitis B (HBV-CH) as determined by ELISA using monoclonal anti-human AFP (mAb-AFP). The differential expression of sialic acid linkage was observed in HBV-CH and HCV-LC by ELISA; the former bound strongly with Sambucus nigra agglutinin (SNA), which has exclusive binding specificity for NeuAcα2-6-, whereas HCV-LC reacted preferably with Maackia amurensis agglutinin (MAA) which recognizes NeuAcα2-3-. There was significantly high glycan branching in HBV-LC and HCV-LC in comparison to controls as illustrated by concanavalin A. This was further confirmed by Phaseolus vulgaris erythroagglutinin (E-PHA) and Datura stramonium agglutinin (DSA). Enhanced fucosylation of AFP was observed in HBV-LC, HCV-LC and HCC patients by ELISA using fucose binding Aleuria aurantia lectin; however, maximum binding was found in HCC. Fucosylation with α1-6 linkage was further confirmed by Lens culinaris agglutinin (LCA). From the above results it is concluded that the changes in concentration of AFP, differential expression of sialic acid, increase of glycan branching and fucosylation have a prognostic value of hepatitis and it could be possible that lectin-based assay with AFP can aid in diagnosis of hepatitis diseases besides clinical examination and routine laboratory investigation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

AFP:

Alpha-fetoprotein

AGP:

Alpha1- acid glycoprotein

ALL:

Aleuria aurantia lectin

AST:

Aspartate aminotransferase

ALT:

Alanine aminotransferase

ALP:

Alkaline phosphatase

ConA:

Concanavalin A

HBV-CH:

Chronic hepatitis B

HBV-LC:

Hepatitis B cirrhosis

HCV-LC:

Hepatitis C cirrhosis

MAA:

Maackia amurensis agglutinin

SNA:

Sambucus nigra agglutinin

Tf:

Transferrin

References

  1. Fattovich, G., Stroffolini, T., Zagni, I., Donato, F.: Hepatocellular carcinoma in cirrhosis: incidence and risk factors. Gastroenterol. 127, S35–50 (2004)

    Article  Google Scholar 

  2. Sheth, S.G., Flamm, S.L., Gordon, F.D., Chopra, S.: AST/ALT ratio predicts cirrhosis in patients with chronic hepatitis C virus infection. Am. J. Gastroenterol. 93, 44–48 (1998)

    Article  PubMed  CAS  Google Scholar 

  3. Williams, A.L., Hoofnagle, J.H.: Ratio of serum aspartate to alanine aminotransferase in chronic hepatitis. Relationship to cirrhosis. Gastroenterol. 95, 734–739 (1988)

    CAS  Google Scholar 

  4. Turner, G.A.: N-glycosylation of serum proteins in disease and its investigation using lectins. Clin. Chim. Acta 208, 149–171 (1992)

    Article  PubMed  CAS  Google Scholar 

  5. Biou, D., Wieruszeski, J.M., Konan, D., Fournet, B., Durand, G.: Hyperfucosylation of α1-acid glycoprotein during cirrhosis. Prog. Clin. Biol. Res. 300, 215–218 (1989)

    PubMed  CAS  Google Scholar 

  6. Thompson, S., Matta, K.L., Turner, G.A.: Changes in fucose metabolism associated with heavy drinking and smoking: a preliminary report. Clin. Chim. Acta 201, 59–64 (1991)

    Article  PubMed  CAS  Google Scholar 

  7. Mann, A.C., Record, C.O., Self, C.H., Turner, G.A.: Monosaccharide composition of haptoglobin in liver diseases and alcohol abuse: large changes in glycosylation associated with alcoholic liver disease. Clin. Chim. Acta 227, 69–78 (1994)

    Article  PubMed  CAS  Google Scholar 

  8. Kondo, M., Hada, T., Fukui, K., Iwasaki, A., Higashino, K., Yasukawa, K.: Enzyme-linked immunosorbent assay (ELISA) for Aleuria aurantia lectin-reactive serum cholinesterase to differentiate liver cirrhosis and chronic hepatitis. Clin. Chim. Acta 243, 1–9 (1995)

    Article  PubMed  CAS  Google Scholar 

  9. Hada, T., Kondo, M., Yasukawa, K., Amuro, Y., Higashino, K.: Discrimination of liver cirrhosis from chronic hepatitis by measuring the ratio of Aleuria aurantia lectin-reactive serum cholinesterase to immunoreactive protein. Clin. Chim. Acta 281, 37–46 (1999)

    Article  PubMed  CAS  Google Scholar 

  10. Mondal, G., Chatterjee, U., Das, H.R., Chatterjee, B.P.: Enhanced expression of α1-acid glycorptein and fucosylation in hepatitis B patients provides an insight into pathogenesis. Glycoconj. J. 26, 1225–1234 (2009)

    Article  PubMed  CAS  Google Scholar 

  11. Crandall, B.F.: Alpha-fetoprotein: a review. CRC Crit. Rev. Clin. Lab. Sci. 15, 127–185 (1981)

    Article  CAS  Google Scholar 

  12. Mizejewski, G.J.: New insights into AFP structure and function: potential biomedical applications. In: Mizejewski, G.J., Porter, I.H. (eds.) Alpha-fetoprotein and congenital disorders, pp. 5–34. Academic, Orlando (1985)

    Google Scholar 

  13. Deutsch, H.F.: Chemistry and biology of α-fetoprotein. Adv. Cancer Res. 56, 253–312 (1991)

    Article  PubMed  CAS  Google Scholar 

  14. Gitlin, D., Perricelli, A., Gitlin, G.M.: Synthesis of AFP by liver, yolk sac and GI tract of the human conception. Cancer Res. 32, 979–982 (1972)

    PubMed  CAS  Google Scholar 

  15. Gitlin, D.: Normal biology of α fetoprotein. Ann. NY Acad. Sci. 259, 7–16 (1975)

    Article  PubMed  CAS  Google Scholar 

  16. Ball, D., Rose, E., Alpert, E.: Alpha-fetoprotein levels in normal adults. Am. J. Med. Sci. 303, 157–159 (1992)

    Article  PubMed  CAS  Google Scholar 

  17. Sizaret, P., Martel, N., Tuyns, A., Reynaud, S.: Mean alpha-fetoprotein values of 1, 333 males over 15 years by age groups. Digestion. 15, 97–103 (1977)

    Article  PubMed  CAS  Google Scholar 

  18. Aoyagi, Y., Suzuki, Y., Isemura, M., Soga, K., Ozaki, T., Ichida, T., Inoue, K., Sasaki, H., Ichida, F.: Differential reactivity of α-fetoprotein with lectins and evaluation of its usefulness in the diagnosis of hepatocellular carcinoma. Gann 75, 809–815 (1984)

    PubMed  CAS  Google Scholar 

  19. Chan, D.W., Miao, Y.C.: Affinity chromatographic separation of alpha-fetoprotein variants: development of a mini-column procedure and application to cancer patients. Clin. Chem. 32, 2143–2146 (1986)

    PubMed  CAS  Google Scholar 

  20. Breborowicz, J., Mackiewicz, A., Breborowicz, D.: Microheterogeneity of alpha-fetoprotein in patient serum as demonstrated by lectin afftno-electrophoresis. Scand. J. Immunol. 14, 15–20 (1981)

    Article  PubMed  CAS  Google Scholar 

  21. Buamah, P.K., Cornell, C., Cassells-Smith, A.J., Skillen, A.W.: Differential reactivity of α-fetoprotein with lectins and its usefulness in the diagnosis of various liver diseases. Clin. Chim. Acta 168, 69–73 (1987)

    Article  PubMed  CAS  Google Scholar 

  22. Toftager-Larsen, K., Peterson, P.L., Norgood-Pedersen, B.: Carbohydrate heterogeneity of human alpha-fetoprotein: oncodevelopmental aspects. In: Bog-Hanson, T.C. (ed.) Lectins-biology, biochemistry, clinical biochemistry, vol. 1, pp. 283–292. Walter de Gruyter, Berlin (1981)

    Google Scholar 

  23. Ishiguro, T., Sakaguchi, H., Fukui, M.: Serum alpha-fetoprotein subfractions in hepatic malignancies identified by different reactivities with concanavalin A, lentil lectin or phytohemagglutinin-E. Jpn J. Surg. 16, 16–21 (1986)

    Article  PubMed  CAS  Google Scholar 

  24. Buamah, P.K., Gibb, I., Bates, G., Mitford, W.A.: Serum alpha fetoprotein heterogeneity as a means of differentiation between primary hepatocellular carcinoma and hepatic secondaries. Clin. Chim. Acta 139, 313–316 (1984)

    Article  PubMed  CAS  Google Scholar 

  25. Green, E.D., Baenziger, J.U.: Characterization of oligosaccharides by lectin affinity high-performance liquid chromatography. Trends Biochem. Sci. 14, 168–172 (1989)

    Article  PubMed  CAS  Google Scholar 

  26. Ishibashi, K., Nishikawa, A., Hayashi, N., Kasahara, A., Sato, N., Fujii, S., Kamada, T., Taniguchi, N.: N-Acetylglucosaminyltransferase in human serum and liver and hepatoma tissues: increased activity in liver cirrhosis and hepatoma patients. Clin. Chim. Acta 185, 325–332 (1989)

    Article  PubMed  CAS  Google Scholar 

  27. Miyoshi, E., Noda, K., Yamaguchi, Y., Inoue, S., Ikeda, Y., Wang, W., Ko, J.H., Uozumi, N., Li, W., Taniguchi, N.: The α 1-6-fucosyltransferase gene and its biological significance. Biochim. Biophys. Acta 1473, 9–20 (1999)

    PubMed  CAS  Google Scholar 

  28. Breborowicz, J., Mackiewicz, A., Breborowicz, D.: Microheterogeneity of α-fetoprotein in patient serum as demonstrated by lectin affino-electrophoresis. Scand. J. Immunol. 14, 15–20 (1981)

    Article  PubMed  CAS  Google Scholar 

  29. Miyazaki, J., Endo, Y., Oda, T.: Lectin affinities of α-fetoprotein in liver cirrhosis, hepatocellular carcinoma and metastatic liver tumor. Acta Hepatol. Jpn. 22, 1559–1568 (1981)

    CAS  Google Scholar 

  30. Kornfeld, S., Reitman, M.L., Kornfeld, R.: The carbohydrate binding specificity of pea and lentil lectins. J. Biol. Chem. 256, 6633–6640 (1981)

    PubMed  CAS  Google Scholar 

  31. Taketa, K., Ichikawa, E., Taga, H., Hirai, H.: Antibody-affinity blotting, a sensitive technique for the detection of α-fetoprotein separated by lectin affinity electrophoresis in agarose gels. Electrophoresis 6, 492–497 (1985)

    Article  CAS  Google Scholar 

  32. Taketa, K.: A tetrazolium method for peroxidase staining; application to the antibody-affinity blotting of α-fetoprotein in cancer diagnosis. Electrophoresis 8, 409–414 (1987)

    Article  CAS  Google Scholar 

  33. Aoyagi, Y., Isemura, Y., Yoshizawa, Z., Suzuki, Y., Sekine, C., Ono, T., Ichida, F.: Fucosylation of serum α-fetoprotein with primary hepatocellular carcinoma. Biochim. Biophys. Acta 830, 217–223 (1985)

    Article  PubMed  CAS  Google Scholar 

  34. Suzuki, Y., Aoyagi, Y., Muramatsu, M., Sekine, C., Isemura, M., Ichida, F.: Close topographical relationship in α-fetoprotein (AFP) between a Lens culinaris binding glycan and the epitope recognized by AFP-reactive monoclonal antibody, 18H4. Br. J. Cancer 55, 147–152 (1987)

    Article  PubMed  CAS  Google Scholar 

  35. Aoyagi, Y., Suzuki, Y., Igarashi, K., Saitoh, A., Oguro, M., Yokota, T., Mori, S., Nomoto, M., Isemura, M., Asakura, H.: The usefulness of simultaneous determinations of glucosaminylation and fucosylation induces of α-fetoprotein in the differential diagnosis of neoplastic diseases of the liver. Cancer 67, 2390–2394 (1991)

    Article  PubMed  CAS  Google Scholar 

  36. Taketa, K., Sekiya, C., Namiki, M., Akamatsu, K., Ohta, Y., Endo, Y., Kosaka, K.: Lectin-reactive profiles of α-fetoprotein characterizing hepatocellular carcinoma and related conditions. Gastroenterol. 99, 508–518 (1990)

    CAS  Google Scholar 

  37. Taketa, K., Endo, Y., Sekiya, C., Tanikawa, K., Taga, H., Satomura, S., Matsuura, S., Kawai, T., Hirai, H.: A collaborative study for the evaluation of lectin-reactive α-fetoprotein in early detection of hepatocellular carcinoma. Cancer Res. 53, 5419–5423 (1993)

    PubMed  CAS  Google Scholar 

  38. Sato, Y., Nakata, K., Kato, Y., Shima, M., Ishii, N., Koji, T., Taketa, K., Endo, Y., Nagataki, S.: Early recognition of hepatocellular carcinoma based on altered profiles of α-fetoprotein. N. Engl. J. Med. 328, 1802–1806 (1993)

    Article  PubMed  CAS  Google Scholar 

  39. Shiraki, K., Takase, K., Tameda, Y., Hamada, M., Kosaka, Y., Nakano, T.: A clinical study of lectin-reactive α -fetoprotein as an early indicator of hepatocellular carcinoma in the follow- up of cirrhotic patients. Hepatology 22, 802–807 (1995)

    Article  PubMed  CAS  Google Scholar 

  40. Yamashita, F., Tanaka, M., Satomura, S., Tanikawa, K.: Prognostic significance of Lens culinaris agglutinin-reactive α-fetoprotein in small hepatocellular carcinomas. Gastroenterol. 111, 996–1001 (1996)

    Article  CAS  Google Scholar 

  41. Aoyagi, Y., Isokawa, O., Suda, T., Watanabe, M., Suzuki, Y., Asakura, H.: The fucosylation index of α-fetoprotein as a possible prognostic indicator for patients with hepatocellular carcinoma. Cancer 83, 2076–2082 (1998)

    Article  PubMed  CAS  Google Scholar 

  42. Ajdukiewicz, A.B., Kelleher, P.C., Krawittb, E.L., Waltersc, C.J.P., Masonc, P.B., Koffd, R.S., Bilanger, L.: α-Fetoprotein glycosylation is abnormal in some hepatocellular carcinomas, including white patients with a normal α -fetoprotein concentration. Cancer Lett. 74, 43–50 (1993)

    Article  PubMed  CAS  Google Scholar 

  43. Di Bisceglie, A.M.: Hepatocellular carcinoma: molecular biology of its growth and relationship to hepatitis B virus infection. Med. Clin. North Am. 73, 985–997 (1989)

    PubMed  Google Scholar 

  44. Block, T.M., Mehta, A.S., Fimmel, C.J., Jordan, R.: Molecular viral oncology of hepatocellular carcinoma. Oncogene 22, 5093–5107 (2003)

    Article  PubMed  CAS  Google Scholar 

  45. Marrero, J.A.: Hepatocellular carcinoma. Curr. Opin. Gastroenterol. 22, 248–253 (2006)

    Article  PubMed  Google Scholar 

  46. Sallie, R., Di Bisceglie, A.M.: Viral hepatitis and hepatocellular carcinoma. Gastroenterol. Clin. North Am. 23, 567–579 (1994)

    PubMed  CAS  Google Scholar 

  47. Lok, A., McMahon, B.: Chronic hepatitis B. Hepatology 34, 1225–1241 (2001)

    Article  PubMed  CAS  Google Scholar 

  48. El Serag, H.B., Mason, A.C., Key, C.: Trends in survival of patients with hepatocellular carcinoma between 1977 and 1996 in the United States. Hepatology 33, 62–65 (2001)

    Article  PubMed  CAS  Google Scholar 

  49. Sarbah, S.A., Gramlich, T., Younoszai, A., Osmack, P., Goormastic, M., Grosso, L., Cooper, J.N., Di Bisceglie, A., Seneca, R., Younossi, Z.M.: Risk factors for hepatocellular carcinoma in patients with cirrhosis. Dig. Dis. Sci. 49, 850–853 (2004)

    Article  PubMed  CAS  Google Scholar 

  50. Jezequel, M., Seta, N.S., Corbic, M.M., Jeanne, M.F., Genevieve, M.D.: Modifications of Concanavalin A patterns of α1 acid glycoprotein and α-2-HS glycoprotein in alcoholic liver disease. Clin. Chim. Acta 176, 49–57 (1988)

    Article  PubMed  CAS  Google Scholar 

  51. Spik, G., Debruyne, V., Montreuil, J.: Alterations of the carbohydrate structure of human serotransferrin in liver diseases. In: Popper, H., Reutter, W. (eds.) Structural carbohydrates in the liver, pp. 477–483. MTP, Basel (1983)

    Google Scholar 

  52. Hachulla, E., Laine, A., Hendouin, V., Fournier, C., Maury, F., Mathieu, C., Pruvot, F.R., Declerck, N., Paris, J.C., Degand, P.: Variations in the glycoforms of serum α-antichymotrypsin in liver diseases and after liver transplantation. Clin. Sci. 82, 439–446 (1992)

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This study was financially supported by a research grant (BT/PR4462/BRB/10/350/2003) from Department of Biotechology, Government of India, New Delhi to B.P.C. The authors thank sincerely Mr. Suneel Arora, Department of Hepatology, Post Graduate Institute of Medical Education and Research, Chandigarh for collection of the serum samples.

Author information

Author notes

  1. Urmimala Chatterjee

    Present address: Department of Anatomy, Physiology and Biochemistry, Swedish University of Agriculture Science, Uppsala, Sweden

Authors and Affiliations

  1. Department of School of Management and Science, West Bengal University of Technology, Salt lake, Kolkata, 700064, India

    Gautam Mondal, Urmimala Chatterjee & Bishnu Pada Chatterjee

  2. Department of Hepatology, Post Graduate Institute of Medical Education and Research, Chandigarh, 160012, India

    Yogesh K. Chawla

Authors
  1. Gautam Mondal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Urmimala Chatterjee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yogesh K. Chawla
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bishnu Pada Chatterjee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bishnu Pada Chatterjee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mondal, G., Chatterjee, U., Chawla, Y.K. et al. Alterations of glycan branching and differential expression of sialic acid on alpha fetoprotein among hepatitis patients. Glycoconj J 28, 1–9 (2011). https://doi.org/10.1007/s10719-010-9316-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-010-9316-z

Keywords

Search

Navigation