Search PubMed⌕ Search

Biomedical subjects

T Waks

Publications and source records attributed to T Waks.

26 records · Page 2Linked to original sources
PubMed1981

The production of monoclonal antibodies against glomerular and other antigens of the human nephron.

During our studies on human kidney tubular antigens we have applied the technique of cell fusion for the preparation of monoclonal antibodies. For this purpose, plasma membranes were prepared from human kidney cortex by homogenization, fractionated on density gradients and selected according to brush border marker enzyme activity. Spleen cells from Balb/c mice hyperimmunized with plasma membranes were fused (PEG) with NS1 plasmocytoma cells by standard procedure. Culture supernatants were tested for presence of specific antibodies by indirect immunofluorescence with fluorescein-conjugated rabbit anti-mouse Fab antibodies on human kidney slices. In two fusions (210 wells), 70 positive hybrids were found secreting antibodies for a variety of antigens in the kidney. Most of them were directed against tubular antigens. In addition, as a by-product, we detected hybridomas which secreted antibodies specific for antigens in other parts of the nephron, such as glomeruli, blood cloning, and monoclonal antibodies were produced in large amounts from ascitic fluid. Some of these antibodies are specific for antigens of the basement membrane, others for antigens of the mesangium. Some recognize antigens present on glomeruli alone, others recognize antigens present on glomeruli and tubules or on glomeruli and blood vessels. We are convinced that the new immunological technique will yield better information on the antigenic microstructure of the nephron. In addition, the monoclonal and, by definition, monospecific antibodies might be useful for diagnostic purposes: recognition and quantitation of the corresponding antigens in the serum and/or urine of patients suffering from kidney diseases.

Antibodies, Monoclonal↗
PubMed1980

Generation of hybridomas secreting murine reaginic antibodies of anti-DNP specificity.

High frequency of hybridoma lines secreting large amounts of reaginic anti-DNP antibodies were generated by the fusion of splenic lymphocytes enriched by adoptive transfer of immune cells and the NSI plasmacytoma cell line. The murine IgE molecule binds DNP with high affinity and was specifically purified by immunoabsorption. The 190,000 dalton molecule is composed of a lambda-light chain and an epsilon-heavy chain of an apparent m.w. of about 90,000. Absorption studies with anti-class-specific antisera suggest that some of the hybrid clones secrete also a mixed molecule consisting of lambda- and kappa-light chains in addition to the epsilon-heavy chain. The monoclonal reaginic antibody binds to mast cells or rat basophilic leukemia cells and, upon binding of DNP-protein, triggers an anaphylactic reaction.

Animals↗
PubMedDate not supplied

Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells.

The antitumor specificity of T cells can be induced by gene transfer using a recently developed therapeutic approach (T body). In this work, we genetically conferred anticarbohydrate specificity onto T cells using the variable regions of monoclonal antibody MLuC1, which binds the Lewis(Y) (LeY) tumor-associated antigen that is overexpressed on several human carcinomas. The variable regions of MLuC1, which are in a single-chain Fv (ScFv) configuration, were cloned and spliced in a eukaryotic expression vector with both the gene encoding the signal-transducing gamma-chain of the human Fcgamma receptor and a flexible hinge domain. The chimeric ScFv-gamma gene was expressed in a murine cytotoxic T-cell hybridoma. Transfectants receiving vector only served as a negative control (mock). Screening for functional transfectants was carried out using a tumor growth inhibition assay. The soluble form of MLuC1 ScFv was recovered from bacteria periplasm and tested for binding to LeY-expressing cells by the fluorescence-activated cell sorter analysis. Despite the low binding ability of the soluble MLuC1 ScFv, 7 of 13 genetically engineered cytotoxic T lymphocyte clones inhibited the growth of LeY-positive cells and did not affect growth of LeY-negative cells. None of the mock clones tested specifically inhibited tumor growth. These data indicate that, by chimeric MLuC1 ScFv-gamma gene transfer, it is possible to confer anticarbohydrate specificity onto T cells and extend the applicability of the T-body approach to tumor-associated antigens that are naturally not recognized by T cells.

Amino Acid Sequence↗