Immunodominant role of CCHA subunit of Concholepas hemocyanin is associated with unique biochemical properties

María Inés Becker, Alejandra Fuentes, Miguel Del Campo, Augusto Manubens, Esteban Nova, Harold Oliva, Fernando Faunes, María Antonieta Valenzuela, Marcelo Campos-Vallette, Alvaro Aliaga, Jorge Ferreira, Alfredo E. De Ioannes, Pablo De Ioannes, Bruno Moltedo

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31 Citations (Scopus)


Hemocyanin, the oxygen transporter metallo-glycoprotein from mollusks, shows strong relationship between its notable structural features and intrinsic immunomodulatory effects. Here we investigated the individual contribution of CCHA and CCHB subunits from Concholepas hemocyanin (CCH) to in vivo humoral immune response and their pre-clinical evaluation as immunotherapeutic agent in a mice bladder cancer model, in relation to their biochemical properties. To this end, subunits were purified and well characterized. Homogeneous subunits were obtained by anionic exchange chromatography, and its purity assessed by electrophoretic and immunochemical methods. While each CCH subunit contains eight functional units showing partial cross reaction, the vibrational spectral analysis showed several spectral differences, suggesting structural differences between them. In addition, we demonstrated differences in the carbohydrate content: CCHA had a 3.6% w/w sugar with both N- and O-linked moieties. In turn, CCHB had a 2.5% w/w sugar with N-linked, while O-linked moieties were nearly absent. Considering these differences, it was not possible to predict a priori whether the immunogenic and immunotherapeutic properties of subunits might be similar. Surprisingly, both subunits by itself induced a humoral response, and showed an antitumor effect in the bladder carcinoma cell line MBT-2. However, when immunologic parameters were analyzed, CCHA showed better efficiency than CCHB. No allergic reactions or any toxic effects were observed in mice treated with CCHA, sustaining its potential therapeutic use. Our study supports that CCHA subunit accounts for the most important features involved in the immunogenicity of CCH, such as better hydrophilicity and higher content of carbohydrates.

Original languageEnglish
Pages (from-to)330-339
Number of pages10
JournalInternational Immunopharmacology
Issue number3
Publication statusPublished - Mar 2009


  • Bladder cancer
  • CCHA and CCHB subunits characterization
  • Concholepas concholepas
  • Hemocyanin
  • Immunogen
  • Immunotherapy

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology


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