A scFv Phage Display Mini Library Generated from the Immunoglobulin Repertoire of Breast Medullary Carcinoma Infiltrating B Lymphocytes

Abstract

(VL) variable region genes was performed. A single chain Fv phage display mini-library was generated from the selected VH and VL genes.

Novel Ganglioside Antigen Identified by B Cells in Human Medullary Breast Carcinomas: The Proof of Principle Concerning the Tumor-Infiltrating B Lymphocytes

“…The construction of the scFv minilibrary was conducted, as described (27,28), by selecting V H and V genes and amplified with specific primers based upon DNA sequence analysis. Assembly reactions of the selected rearranged Ig V region H (26VH) and L chain (32V) genes were conducted by a three-step PCR amplification, using a linker peptide (Gly 4 Ser 3 ) coding sequence.…”

“…Our model system for characterizing the expressed Ig repertoire (22,27,28) was established from high grade MBC because this has the highest B cell infiltration (29). A combinatorial single chain Fv (scFv) minilibrary from selected V H and V region genes was generated and tested thereafter for breast tumor cell-binding capacity.…”

Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells

“…Early evidence that tumor-infiltrating B lymphocyte (TIL-B)-derived antibodies may also recognize tumor cells was obtained in the following ways: by production of human hybridomas derived from TIL, able to secrete tumor-specific antibodies [ 19 , 20 ]; B cell expansion of TIL from human tumor biopsies [ 21 ]; B cell expansion of melanoma-derived TIL, and following cloning of the scFv antibody with specific melanoma reactivity from single B cell clone [ 22 ]; and subcutaneous transplantation of human lung cancer tissue in immunodeficient mice [ 23 , 24 ], all of which suggest a specific function of TIL-B in the tumor. Recently, a rare type of breast cancer, classified as medullary carcinoma (MCB, medullary carcinoma of breast), characterized by strong lymphoplasmacytic infiltrates correlated with improved prognosis and patient survival, and cervical carcinoma, were investigated to understand the nature of tumor-infiltrated B lymphocytes through analysis of TIL-derived Ig repertoire [ 25 - 28 ]. A study of the molecular structure of variable antibody regions gave evidence of antigen-driven humoral immune responses in medullary breast carcinomas, as well as in cervical tumors.…”

The Ectopic Germinal Centre Response in Autoimmune Disease and Cancer

“…Some TAAs appear to overcome tolerance and induce a natural immune response as a result of mutation or altered expression; humoral immune responses to these antigens in breast cancer patients are associated with better early disease stage-specific survival (Angelopoulou et al, 2000;Visco et al, 2000;von Mensdorff-Pouilly et al, 2000) and anti-MUC-1 antibodies are cytotoxic to tumour cells (Snijdewint et al, 2001). TAA-specific tumour infiltrating (TIL) B-lymphocytes and recombinant antibodies have been isolated from both tumour (Kotlan et al, 2000; and lymph nodes (Petrarca et al, 1999;Rothe et al, 2004) of medullary and ductal carcinoma patients, showing that they are responding specifically to the tumour. Evasion of the immune response by the tumour can be overcome by passive immunotherapy or active immunisation regimes.…”