ABSTRACT
The production of alloreactive cytotoxic T-lymphocytes (CTL) for therapy of recurrent brain tumours was performed in the CELLMAX artificial capillary system composed of cell-culture modules containing cellulose acetate or cuprammonium rayon hollow fibres. Lymphocytes, obtained from the brain-tumour patient to be used for sensitization, were stimulated with OKT3 (anti-CD3) and interleukin-2 (IL-2) and inoculated into the extracapillary space of artificial capillary modules. For allogeneic CTL production, the expanded patient lymphocytes were harvested, irradiated and placed into a second artificial capillary system with allogeneic lymphocytes from a healthy donor. In a one-way mixed lymphocyte reaction, CTL developed in the presence of low-concentration IL-2 (60 i.u. of IL-2/ml). In 18-21 days the cell preparation usually displayed a predominantly CD3+, CD8+ phenotype, which consorted with the dual-labelled CD8/CD11a markers used to identify CTL. Chromium (Cr)-release assays demonstrated lysis of patient tumour in relation to allogeneic glioma; the response observed in cold-target-inhibition assays confirmed lysis of the relevant tumour by the CTL.
Subject(s)
Brain Neoplasms/therapy , Immunotherapy, Adoptive , Neoplasm Recurrence, Local/therapy , T-Lymphocytes, Cytotoxic/immunology , Antibodies, Monoclonal/immunology , Biocompatible Materials/therapeutic use , CD3 Complex/genetics , CD3 Complex/immunology , CD8 Antigens/genetics , CD8 Antigens/immunology , Cells, Cultured , Cellulose/analogs & derivatives , Cellulose/therapeutic use , Chromium/metabolism , Gene Expression Regulation, Neoplastic/genetics , Gene Expression Regulation, Neoplastic/immunology , Glioma/metabolism , Glioma/pathology , Glioma/therapy , Humans , Immunosuppressive Agents/pharmacology , Interleukin-2/pharmacology , Lymphocyte Activation/drug effects , Lymphocyte Culture Test, Mixed , Lymphocyte Function-Associated Antigen-1/genetics , Lymphocyte Function-Associated Antigen-1/immunology , Membranes, Artificial , Muromonab-CD3/pharmacology , Phenotype , T-Lymphocytes, Cytotoxic/cytology , T-Lymphocytes, Cytotoxic/radiation effects , Tumor Cells, CulturedABSTRACT
A colorimetric enzyme immunoassay (EIA) method for detection of Salmonella in foods has been compared to the AOAC colorimetric monoclonal EIA screening method (986.35, 15th ed.; 46.B21-46.B29, 14th ed.). The assays use the same monoclonal antibodies and have similar reactivity toward Salmonella. However, the new assay uses antibody-coated microtiter wells instead of coated magnetic beads to capture Salmonella antigens. Compared with the bead assay, the coated-well assay format requires significantly less time to complete, and was consistently able to detect lower levels of Salmonella in mixed culture. Compared to the standard AOAC culture method for food samples, the plate assay was as productive. No false negatives were obtained by the immunoassay; the false negative rate was 1.1% by the culture method. The rate of agreement between the 2 methods was 99.1%. The official final action bead assay method for Salmonella in foods, 986.35, and the same assay for use with low-moisture foods, 987.11, have been modified official first action to use antibody-coated microtiter strip-wells.
