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AssigneeUniversity of Illinois Foundation
InventorsMarius C. Teodorescu, Eugene P. Mayer, Sheldon Dray
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A method for identification of cell subpopulations in a population of cells which are morphologically alike, based on the ability of selected strains of bacteria to bind to the cell surface when they are brought in close contact. Selected strains of naturally occurring or artificially obtained bacteria of different species or genuses, having an affinity to bind to different subpopulations of a cell mixture, are brought into contact by centrifugation or the like with a mixture of cells such as leukocytes, the mixture is resuspended and the cells having bacteria attached are identified under the microscope. The method is useful particularly for the identification of B cells, several subpopulations of T cells, and the differentiation among leukemic cell populations.
A method for detecting, identifying and enumerating a subpopulation of leukocytes in a mixture of morphologically indistinguishable leukocytes having different membrane characteristics, which method comprises bringing said mixture into close contact with a previously selected strain of bacteria having an affinity for binding only to the cells of said subpopulation by a mechanism not involving the binding sites of any immunoglobulins on the surface of said leukocytes, and observing the presence of said bacteria bound to said subpopulation as a morphologically distinct marker for identification and enumeration of said subpopulation.
A method for quantifying each of a plurality of subpopulations of morphologically undistinguishable lymphocytes in a mixture thereof which method comprises contacting said mixture with a plurality of bacteria selected from the group consisting of Arizona hinshawii, ATTC 31241;
A method for producing mutants of bacteria having a propensity for binding spontaneously to a specific subpopulation of leukocytes which comprises treating with a mutagenic agent a non-binding strain of bacteria known to require a particular nutrient for growth, bringing the treated strain into contact with said subpopulation of leukocytes to permit the association of mutants of said bacteria with said subpopulation of leukocytes, washing to remove non-binding bacteria, plating the leukocytes onto a growth medium lacking the particular nutrient required for the growth of said strain, collecting any bacteria that grow into colonies, and selecting the clones which recognize said subpopulation of leukocytes.
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