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

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.

Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor

“…Antibodies with binding capacity to sialylated glycosphingolipid structures can neutralize shed gangliosides. This helps to restore the antitumor reaction that is diminished by shed tumor-associated molecules (31) and might influence the strength of the immune response in general.…”

Challenging tumour immunological techniques that help to track cancer stem cells in malignant melanomas and other solid tumours

“…Our original model system was based on a rare type of invasive ductal breast carcinomas, showing extensive B cell infiltration and other specific characteristics [ 17 , 18 ]. Additional relevant information on the antibody repertoire of patients with breast cancer [ 19 ] or malignant melanomas [ 13 ] supported the further potentials of this approach. Comparative DNA sequence analyses seem to be an effective effort helping to reveal human antibody variable region genes of interest.…”

“…We approached the problem with another theory. Taking into account that „Nature must know the answer to cancer”, we have developed a novel strategy: our molecular genetic technology [ 10 , 11 ], essential findings on cancer associated antigens [ 12 ] and a complex tumorimmunological project [ 13 ] helped us to reveal cancer targeting molecules. B lymphocites accumulated in the tumor microenvironment, the so called tumor infiltrating B cells (TIL-B) that can be found in breast cancers and other solid tumors [ 14 , 15 ], can be analysed in terms of their antibody repertoire.…”

Notch4+ cancer stem‐like cells promote the metastatic and invasive ability of melanoma

“…As is well known, melanoma is a highly lethal malignant tumor with a high rate of metastasis. Once metastatic, the outcomes for patients are poor . Clarification of the mechanisms that underlie tumor metastasis is essential for the improvement in cancer survival of individuals with cancer.…”