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Toll-like receptor 4 gene polymorphism influences dendritic cell in vitro function and clinical outcomes in vaccinated melanoma patients

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Abstract

Toll-like receptor 4 (TLR4) is expressed on dendritic cells (DCs), sensing environmental danger molecules that induce their activation and maturation. Recently, we reported a method for the production of therapeutic DCs against melanoma, called tumor antigen-presenting cells (TAPCells), using a heat-shocked allogeneic melanoma cell lysate (TRIMEL) as an activation factor and antigen provider. Since TRIMEL contains endogenous TLR4 ligands, we evaluated the role of TLR4 in TAPCells differentiation by antibody neutralization and the association of a Tlr4 polymorphism (896A/G) (Asp299Gly), determined by PCR–RFLP, with the in vitro activation capacity and the clinical outcome of TAPCells-vaccinated patients. Antibody blocking of monocyte TLR4 inhibited surface expression, determined by flow cytometry, of the major histocompatibility complex class I, CCR7, CD80, CD83 and CD86 on TAPCells, reduced interleukin (IL)-6 and tumor necrosis factor -α gene expression evaluated by qRT-PCR, and also inhibited the TAPCells-mediated interferon-γ (IFN-γ) secretion of melanoma-specific CD8+ T cells determined by ELISpot (p < 0.01). Moreover, CD8+ T-cell activation capacity was significantly reduced in TAPCells bearing the TLR4 Asp299Gly receptor (p < 0.05). Finally, TAPCells-vaccinated stage-IV melanoma patients bearing the Tlr4 896G allele showed a shortened post-therapy median survival rate compared with those carrying the Tlr4 896A allele (p < 0.05; log-rank test). Our results indicate that TLR4 is a key receptor for the tumor lysate-mediated in vitro generation of clinically efficient antigen-presenting cells. Further analysis of patients included in different vaccine protocols is necessary for definitively establishing a role for TLR4 polymorphism in clinical responses.

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Acknowledgments

This study was supported by grants from the National Fund for Scientific and Technological Development (FONDECYT 1090238, and 1090243), the Fund for the Promotion of Scientific and Technological Development (FONDEF DO5I10366), and the Millennium Science Initiative from the Ministry for the Economy, Development and Tourism (P09/016-F).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, 8380453, Santiago Chile, Chile

    Andrés Tittarelli, Fermín E. González, Cristián Pereda, Gabriela Mora, Leonel Muñoz, Carlos Saffie, Tamara García, David Díaz, Cristián Falcón, Marcela Hermoso, Mercedes N. López & Flavio Salazar-Onfray

  2. Millennium Institute on Immunology and Immunotherapy, University of Chile, 8380453, Santiago, Chile

    Andrés Tittarelli, Fermín E. González, Cristián Pereda, Gabriela Mora, Tamara García, Cristián Falcón, Mercedes N. López & Flavio Salazar-Onfray

  3. Department of Conservative Dentistry, Faculty of Dentistry, University of Chile, 8380492, Santiago, Chile

    Fermín E. González

  4. Research Support Office, University of Chile Clinical Hospital, 8380453, Santiago, Chile

    Mercedes N. López

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  1. Andrés Tittarelli
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Correspondence to Flavio Salazar-Onfray.

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Tittarelli, A., González, F.E., Pereda, C. et al. Toll-like receptor 4 gene polymorphism influences dendritic cell in vitro function and clinical outcomes in vaccinated melanoma patients. Cancer Immunol Immunother 61, 2067–2077 (2012). https://doi.org/10.1007/s00262-012-1268-7

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  • DOI: https://doi.org/10.1007/s00262-012-1268-7

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