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Delivery of immunostimulatory monoclonal antibodies by encapsulated hybridoma cells

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Immunostimulatory monoclonal antibodies are immunoglobulins directed toward surface proteins of immune system cells that augment the immune response against cancer in a novel therapeutic fashion. Exogenous administration of the recombinant humanized immunoglobulins is being tested in clinical trials with agents of this kind directed at a variety of immune-controlling molecular targets. In this study, the encapsulation of antibody-producing hybridoma cells was tested in comparison with the systemic administration of monoclonal antibodies. Hybridomas producing anti-CD137 and anti-OX40 mAb were encapsulated in alginate to generate microcapsules containing viable cells that secrete antibody. Immobilized cells in vitro were able to release the rat immunoglobulin produced by the hybridomas into the supernatant. Microcapsules were implanted by injection into the subcutaneous tissue of mice and thereby provided a platform for viable secreting cells, which lasted for more than 1 week. The pharmacokinetic profile of the rat monoclonal antibodies following microcapsule implantation was similar to that attained following an intraperitoneal administration of the purified antibodies. The rat–mouse hybridoma cells did not engraft as tumors in immunocompetent mice, while they lethally xenografted in immunodeficient mice, if not microencapsulated. The antitumor therapeutic activity of the strategy was studied on established CT26 colon carcinomas resulting in complete tumor eradication in an elevated fraction of cases and strong tumor-specific CTL responses with either anti-CD137 or anti-OX40 producing hybridomas, thus offering proof of the concept. This form of administration permitted combinations of more than one immunostimulatory monoclonal antibody to exploit the synergistic effects such as those known to be displayed by anti-CD137 and anti-OX40 mAb.

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Abbreviations

mAb:

Monoclonal antibody

DC:

Dendritic cell

TNF:

Tumor necrosis factor

NK:

Natural killer

APA:

Alginate-poly-l-lysine-alginate

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Acknowledgments

We are grateful to Drs. Lieping Chen for the kind gift of the 2A hybridoma and Mario Colombo for OX86 cells. Elena Ciordia and Eneko Elizalde are acknowledged for their excellent animal facility management. We received financial support from MEC/MICINN (SAF2005-03131 and SAF2008-03294), Departamento de Educación del Gobierno de Navarra, Departamento de Salud del Gobierno de Navarra (Beca Ortiz de Landázuri), Redes temáticas de investigación cooperativa RETIC (RD06/0020/0065), Fondo de investigación sanitaria (FIS PI060932), European Commission FP7 (ENCITE) and SUDOE-IMMUNONET (FEDER) which supports JD, Fundacion Mutua Madrileña and “UTE for project FIMA”. M S-H received a Ramon y Cajal contract from Ministerio de Ciencia y tecnología and A P a scholarship from FIS.

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Authors and Affiliations

  1. Centro de Investigación Médica Aplicada (CIMA), University of Navarra, CIMA. Av. Pio XII, 55 31008, Pamplona, Spain

    Juan Dubrot, Asis Palazón, Ana Rouzaut, Sandra Hervás-Stubbs & Ignacio Melero

  2. Laboratory of Pharmacy and Pharmaceutical Technology, Networking Biomedical Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, SLFPB-EHU, Faculty of Pharmacy, University of the Basque Country, 01006, Vitoria-Gasteiz, Spain

    Aitziber Portero, Gorka Orive, Rosa María Hernández & Jose Luis Pedraz

  3. Clinica Universitaria, Universidad de Navarra, Pamplona, Spain

    Jose L. Perez-Gracia & Ignacio Melero

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  1. Juan Dubrot
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  2. Aitziber Portero
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  3. Gorka Orive
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  4. Rosa María Hernández
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  6. Ana Rouzaut
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  7. Jose L. Perez-Gracia
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  8. Sandra Hervás-Stubbs
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  9. Jose Luis Pedraz
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  10. Ignacio Melero
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Corresponding authors

Correspondence to Jose Luis Pedraz or Ignacio Melero.

Additional information

I. Melero and J. L. Pedraz equally share credit for senior authorship.

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Dubrot, J., Portero, A., Orive, G. et al. Delivery of immunostimulatory monoclonal antibodies by encapsulated hybridoma cells. Cancer Immunol Immunother 59, 1621–1631 (2010). https://doi.org/10.1007/s00262-010-0888-z

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  • DOI: https://doi.org/10.1007/s00262-010-0888-z

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