Immunologic Research

, Volume 61, Issue 1–2, pp 11–23 | Cite as

The novel panel assay to define tumor-associated antigen-binding antibodies in patients with metastatic melanomas may have diagnostic value

  • Beatrix Kotlan
  • Gabriella Liszkay
  • Miri Blank
  • Orsolya Csuka
  • Timea Balatoni
  • Laszlo Toth
  • Klara Eles
  • Szabolcs Horvath
  • Gyorgy Naszados
  • Judit Olasz
  • Balazs Banky
  • Jozsef Toth
  • Maria Godeny
  • Francesco M. Marincola
  • Miklos Kasler
  • Yehuda Shoenfeld
CUTTING EDGE IN AUTOIMMUNITY

Abstract

We aim to harness the natural humoral immune response by various technologies to get novel biomarkers. A complex antibody analysis in sera and in the tumor microenvironment leads to reveal tumor-specific antibodies. More strategies were introduced to select the most effective one to identify potential tumor antigen-binding capacity of the host. Epstein–Barr virus transformation and cloning with limiting dilution assay, magnetic cell sorting and antibody phage display with further methodological improvements were used in epithelial and neuroectodermal cancers. Column-purified sera of patient with melanoma were tested by immunofluorescence assay, while sera of further melanoma patients were processed for membrane-binding enzyme-linked immunosorbent assay. Some supernatants of selected B cell clones and purified antibodies showed considerable cancer cell binding capacity by immunofluorescence FACS analysis and confocal laser microscopy. Our native tumor cell membrane preparations helped to test soluble scFv and patients’ sera for tumor binder antibodies. A complex tumor immunological study was introduced for patients with melanoma (ethical permission: ETT TUKEB 16462-02/2010); peripheral blood (n = 57) and surgically removed primary or metastatic tumors (n = 44) were gathered and processed at cellular immunological level. The technological developments proved to be important steps forward to the next antibody profile analyses at DNA sequence level. Cancer cell binding of patient-derived antibodies and natural immunoglobulin preparations of pooled plasma product intravenous immunoglobulins support the importance of natural human antibodies. Important cancer diagnostics and novel anticancer strategies are going to be built on these tools.

Keywords

Autoantibody Human antibody Tumor antigen Melanoma 

Abbreviations

ATCC

American Type Cell Collections

EBV

Epstein–Barr virus

GD3

Ganglioside 3 type disialylated glycosphingolipids

ELISA

Enzyme-linked immunosorbent assay

FDA

Food and Drug Administration

JITC

Journal for ImmunoTherapy of Cancer

HJLCT

Harry J Loyd Charitable Trust

HSA

Human serum albumin

IDC

Invasive ductal carcinoma of the breast

IF

Immunofluorescence

IHC

Immunohistochemistry

alphaIFN

Interferon alpha

IVIG

Intracenous immunoglobulin

LDA

Limiting dilution assay

MACS

Magnetic cell sorting

MBC

Medullary breast carcinoma

MTT

2-(4,5-Dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide

ETT-TUKEB

Ministry of Human Resources in Hungary, Hungarian Medical Research Council

NM

Nodular melanoma

PFA

Paraformaldehyde

PBMC

Peripheral blood mononuclear cells

SSM

Superficial spreading melanoma

TAA

Tumor-associated antigen

TIL-B

Tumor infiltrating B cells

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Beatrix Kotlan
    • 1
    • 2
  • Gabriella Liszkay
    • 3
  • Miri Blank
    • 4
  • Orsolya Csuka
    • 5
    • 9
  • Timea Balatoni
    • 3
  • Laszlo Toth
    • 6
  • Klara Eles
    • 2
  • Szabolcs Horvath
    • 2
  • Gyorgy Naszados
    • 7
  • Judit Olasz
    • 5
  • Balazs Banky
    • 8
  • Jozsef Toth
    • 2
  • Maria Godeny
    • 7
    • 9
  • Francesco M. Marincola
    • 10
  • Miklos Kasler
    • 9
  • Yehuda Shoenfeld
    • 4
  1. 1.Molecular Immunology and ToxicologyNational Institute of OncologyBudapestHungary
  2. 2.Center of Surgical and Molecular TumorpathologyNational Institute of OncologyBudapestHungary
  3. 3.OncodermatologyNational Institute of OncologyBudapestHungary
  4. 4.Zabludowitz Center for Autoimmune Diseases, Sheba Medical Center Affiliated to Sackler Faculty of MedicineTel-Aviv UniversityTel AvivIsrael
  5. 5.PathogeneticsNational Institute of OncologyBudapestHungary
  6. 6.OncosurgeryNational Institute of OncologyBudapestHungary
  7. 7.Department of Radiological DiagnosticsNational Institute of OncologyBudapestHungary
  8. 8.County Hospital TatabanyaTatabanyaHungary
  9. 9.National Institute of OncologyBudapestHungary
  10. 10.Sidra Medical and Research CentreDohaQatar

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