Specific antibody synthesis in vitro. I. Technical considerations.
Previously described methods for measuring specific antibody production in vitro are complex and frequently require the addition of a non-specific mitogen and 2 or more separate steps including a lymphocyte culture and then an antibody assay. In this report we present further details regarding a 1-step, non-mitogen requiring assay for specific antibody synthesis combining microculture and enzyme-linked immunoassay techniques. Peripheral blood mononuclear cells from recently immunized subjects are cultured in antigen-coated plastic wells and the specific antibody produced and adhered to the solid-phase antigen is measured in an enzyme-linked immunoassay. The production of specific antibody in vitro did not occur in lymphocyte preparations that were frozen, thawed and incubated, nor did it occur in cultures containing puromycin. T cell separation and remixture experiments indicated that T cells provide significant augmentation under the conditions of this assay. It is apparent that the antibody measured was synthesized in vitro and that cell-cell interactions are operant. The assay may prove useful in the assessment of immune competence and in determining in vitro the effect of certain biologic response modifiers on antibody production.
D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D000918:Antibody Specificity; D000921:Antibody-Producing Cells / Q000276:immunology* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007117:Immunization, Secondary; D008213:Lymphocyte Activation; D008211:Lymphocyte Cooperation; D013601:T-Lymphocytes / Q000276:immunology; D013745:Tetanus Toxoid / Q000008:administration & dosage / Q000276:immunology
D000907:Antibodies, Bacterial; D013745:Tetanus Toxoid