B-lymphoma cells are activated by peptide ligands of the antigen binding receptor or by anti-idiotypic antibody to induce extracellular acidification.
Synthetic peptide ligands specific for the surface immunoglobulin receptor of the human Burkitt's lymphoma cell line SUP-B8, previously identified using phage display libraries, induced apoptosis of the SUP-B8 cells in vitro when administered as dimers or tetramers. The use of synthetic peptide ligands is being explored for immunotherapy of B-cell lymphoma. It will be critical to identify which of the peptide ligands identified are the most active functionally. Using the Cytosensor microphysiometer, SUP-B8 cells and B-lymphoma cells obtained from patients were found to acidify their extracellular environment within minutes of specific activation by surrogate peptide ligands or by anti-idiotype antibodies. This signal was blocked by pretreatment of the lymphoma cells with the tyrosine kinase inhibitor genistein. Treatment of SUP-B8 cells with dimeric and tetrameric specific peptide ligands caused a rapid increase in extracellular acidification rate, which peaked after 10 min at approximately 15 and 20% above basal rates, respectively. These responses were blocked by excess monomeric peptide. To evaluate the ability of different peptide ligands to induce a signal directly on lymphoma cells, thereby establishing their relative affinity to the surface immunoglobulin receptor, acidification rate changes were measured at varying peptide concentrations. The microphysiometer signal correlated with the known relative affinities and antiproliferative potencies of the peptides. This approach is particularly useful for primary tumor cells that cannot be cultured. The signal may be predictive of the efficacy of treatment with synthetic peptide ligands and may be useful in the evaluation of ligands for other cell surface receptors with biological effects on B-lymphoma cells.
Renschler
M F
MF
Division of Oncology, Stanford University School of Medicine, California 94305, USA.
Wada
H G
HG
Fok
K S
KS
Levy
R
R
D000595:Amino Acid Sequence; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism; D004745:Enkephalins / Q000737:chemistry / Q000494:pharmacology*; D005110:Extracellular Space / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007075:Immunoglobulin M / Q000378:metabolism; D008024:Ligands; D016393:Lymphoma, B-Cell / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D011498:Protein Precursors / Q000737:chemistry / Q000494:pharmacology*; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D004745:Enkephalins; D007075:Immunoglobulin M; D008024:Ligands; D010446:Peptide Fragments; D010455:Peptides; D011498:Protein Precursors; D011947:Receptors, Antigen, B-Cell; D011971:Receptors, Immunologic; C056943:peptide B; C050632:polypeptide C