Simultaneous assay for T and B lymphocyte populations and subpopulations
AssigneeBio-Rad Laboratories, Inc.
InventorsMadelyn M. Baran, Dennis M. Bleile
Life of this patent
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Abstract
An assay for simultaneously determining the ratio of B cells and T cells to the total cell population and subpopulations thereof present in a lymphocyte population utilizes excess amounts of B cell binding protein and T cell binding protein bound to solid phase particles. By exposing the particles to the lymphocyte population, rosettes are formed which may be visually distinguished and counted under a microscope, yielding the proportions of B cells and T cells, including any subpopulations for which a specific label was provided. Macrophages may be identified by their ability to ingest the particles.
Claims26
A method for simultaneously determining the relative number of B cells and T cells in a lymphocyte population of a vertebrate, said method employing (1) B cell labels comprising insoluble polymeric particles having a size in the range from 5 to 15 microns bound to B cell binding proteins and (2) T cell labels comprising insoluble polymeric particles having a size in the range from 5 to 15 microns bound to T cell binding protein, at least one of said B cell and T cell labels being covalently coupled to a dye which imparts a color to the particle which is visible under a light microscope, said method comprising:
- incubating the lymphocyte population simultaneously with the B cell labels and T cell labels for a time sufficient for all of the B cells and T cells to specifically form rosettes with the corresponding label;
- and
- counting the number of B cell rosettes and T cell rosettes under a light microscope.
A kit for use in an immunoassay for determining the relative number of B cells and T cells present in an isolated human lymphocyte population, said kit including:
- B cell label comprising anti-(human immunoglobulin) antibody covalently bonded to insoluble polymeric particles having a size in the range from 5 to 15 microns;
- and
- T cell label comprising anti-(T cell) antibody covalently bonded to insoluble polymeric particles having a size in the range from 5 to 15 microns, where at least one of said B cell label and T cell label has been covalently coupled to a dye so that the markers may be distinguishable under a light microscope.
Dependencies are inferred from the claim text.