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Anti-tumor antibody profile analysis to harness the potentials of B cells in melanomas and the natural humoral immune response
  1. Beatrix Kotlan1,
  2. Gabriella Liszkay2,8,9,
  3. Miri Blank3,
  4. Judit Olasz4,
  5. Orsolya Csuka4,8,
  6. Timea Balatoni2,
  7. Kinga Borbola2,
  8. Laszlo Toth5,
  9. Gyorgy Naszados6,
  10. Francesco M Marincola7,
  11. Maria Godeny6,8,9,
  12. Miklos Kasler8,9 and
  13. Yehuda Shoenfeld3
  1. Aff1 grid.419617.c0000000106678064Molecular Immunology and ToxicologyNational Institute of Oncology Budapest Hungary
  2. Aff2 grid.419617.c0000000106678064OncodermatologyNational Institute of Oncology Budapest Hungary
  3. Aff3 grid.413795.d0000000121072845Zabludowicz Center for Autoimmune DiseasesSheba Medical Center Tel Hashomer Israel
  4. Aff4 grid.419617.c0000000106678064PathogeneticsNational Institute of Oncology Budapest Hungary
  5. Aff5 grid.419617.c0000000106678064OncosurgeryNational Institute of Oncology Budapest Hungary
  6. Aff6 grid.419617.c0000000106678064Radiological DiagnosticsNational Institute of Oncology Budapest Hungary
  7. Aff7 grid.467063.00000000403974222SIDRA Medical and Research Center Doha Qatar
  8. Aff8 grid.419617.c0000000106678064Board of DirectorsNational Institute of Oncology Budapest Hungary
  9. Aff9 Univ Med Pharm Tirgu Mures Romania

http://dx.doi.org/10.1186/2051-1426-1-S1-O4

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    Meeting abstracts

    Objectives

    Natural humoral immune response and autoimmune mechanisms have great importance in keeping the balance of tumorimmunity, although it has not yet been fully understood. We aimed to reveal potential anti-tumor immune response by immunoglobulin (Ig) profile analysis of patients with metastatic melanomas.

    Methods

    A complex panel assay has been performed at expressed DNA and protein levels on antibodies originating from patients' peripheral blood (n = 92) or cancerous tissue biopsies (n= 87) (ETT TUKEB 16462- 02/2010). Heavy and light chain immunoglobulin variable gene regions were sequenced and analysed with Vector NTI Advance 11, Bioedit 7.0, ClustalX2.0.11, TreeView 1.6.6 programs using available databases (IMGT, Blast). Patients' sera, purified human Ig preparations and antibody fragments from tumor infiltrating B cells were tested by ELISA, immunofluorescence FACS and confocal laser microscopy.

    Results

    Cluster analysis revealed specific antibody variable region gene subgroups in the VH3 family, amongst which there are the ones with cancer associated antigen binding capacity. The purified immunoglobulin's strong SK-Mel28 melanoma binding potential paralleled with the clinical outcome. Some selected expressed antibody fragments showed tumor associated antigen labelling on melanoma tissues (Fig. 1). A competitive cell membrane ELISA has been standardized for measuring patients' sera in terms of various cancer associated antigen binding capacity. Data are under evaluation concerning their value to predict anti cancer humoral immune response.

    Figure 1
    Figure 1

    Human purified immunoglobulin of patient (a/) and antibody fragments developed (b/) show cancer associated antigen specific binding on SK-Mel 28 cells and melanoma tissues.

    Conclusions

    We could prove the extensive presence of highly tumor associated unique GD3 sialilated glycosphingolipid specific antibody variable regions in patients with melanoma. Our novel panel assay confirmed the potentials of antibody profile analysis in characterizing anti tumor humoral immune response and its worth for diagnostics.

    Acknowledgments

    The Harrry J. Lloyd Charitable Trust Melanoma Research Foundation Award and previous Fulbright No1206103 and OTKA T048933 Grants are acknowledged.