METHODS FOR DETERMINING INTERACTION BETWEEN BIOLOGICAL CELLS
The present invention relates to methods for determining the propensity of cells to interact in a population of cells comprising at least two distinguishable subpopulations of cells. In addition, the present invention provides methods for diagnosing a disease or predisposition to a disease of a cell donor, wherein the method comprises the determination of the propensity of cells to interact in a population of cells obtained from the donor, wherein the population of cells comprises at least two distinguishable subpopulations of cells. The invention also provides methods for determining whether a subject suffering from or predisposed to a disease will respond or is responsive to treatment with a therapeutic agent by determining the alteration of the propensity of cells to interact in a population of ceils obtained from the subject, wherein the population of cells comprises at least two distinguishable subpopulations of cells. The invention also relates to methods for screening for a therapeutic agent comprising the determination whether one or more test substances alter the propensity of cells to interact.
1 . A method for determining the propensity of cells to interact in a population of cells comprising at least two distinguishable subpopulations of cells, wherein the method comprises the following steps:
(a) determination of the number of interactions between cells of a first distinguishable subpopulation and cells of a second distinguishable subpopulation;
(b) determination of the number of interactions in the population of cells by randomly assigning cells comprised in the population of cells to first and second distinguishable subpopulation of cells, wherein the absolute number of cells in the first and second distinguishable subpopulation of cells, respectively, are identical to the number of cells in the first and second distinguishable subpopulation of cells in (a); and
(c) determination of the propensity of cells to interact by dividing (a) through (b), wherein propensity increases with the resulting number.
2 . A population of cells obtained from a cell donor to determine whether the cell donor suffers from a disease or is predisposed to suffer from a disease, wherein the determination comprises the determination of the propensity of cells in the population to interact, wherein the population comprises at least two distinguishable subpopulations of cells and wherein the method comprises the following steps:
(a) determination of the number of interactions between cells of a first distinguishable subpopulation and cells of a second distinguishable subpopulation;
(b) determination of the number of interactions in the population of cells by randomly assigning cells comprised in the population of cells to first and second distinguishable subpopulation of cells, wherein the absolute number of cells in the first and second distinguishable subpopulation of cells, respectively, are identical to the number of cells in the first and second distinguishable subpopulation of cells in (a);
(c) determination of the propensity of cells to interact by dividing (a) through (b), wherein propensity increases with the resulting number; and
(d) determination whether the cell donor has a disease or is predisposed to have a disease based on the propensity of cells to interact.
3 . A method for diagnosing a disease or predisposition to a disease of a cell donor, wherein the method comprises the determination of the propensity of cells to interact in a population of cells obtained from the donor, wherein the population of cells comprises at least two distinguishable subpopulations of cells and wherein the method comprises the following steps:
(a) determination of the number of interactions between cells of a first distinguishable subpopulation and cells of a second distinguishable subpopulation;
(b) determination of the number of interactions in the population of cells by randomly assigning cells comprised in the population of cells to first and second distinguishable subpopulation of cells, wherein the absolute number of cells in the first and second distinguishable subpopulation of cells, respectively, are identical to the number of cells in the first and second distinguishable subpopulation of cells in (a);
(c) determination of the propensity of cells to interact by dividing (a) through (b), wherein propensity increases with the resulting number; and
(d) determination whether the cell donor suffers from a disease or is predisposed to suffer from a disease based on the propensity of cells to interact.
4 . A method for determining whether a subject suffering from or predisposed to a disease will respond or is responsive to treatment with a therapeutic agent by determining the alteration of the propensity of cells to interact in a population of cells obtained from the subject, wherein the population of cells comprises at least two distinguishable subpopulations of cells and wherein the method comprises the following steps:
(a) determination of the number of interactions between cells of a first distinguishable subpopulation and cells of a second distinguishable subpopulation;
(b) determination of the number of interactions in the cell sample by randomly assigning cells comprised in the population of cells to first and second distinguishable subpopulation of cells, wherein the absolute number of cells in the first and second distinguishable subpopulation of cells are identical to the number of cells in the first and second distinguishable subpopulation of cells in (a);
(c) determination of the propensity of cells to interact by dividing (a) through (b), wherein propensity increases with the resulting number; and
(d) determination of the alteration of the propensity of cells to interact by comparing the propensity prior to addition of a therapeutic agent and subsequent to addition of a therapeutic agent and thereby determining whether the subject will respond or is responsive to treatment with the therapeutic agent.
5 . (canceled)
6 . A method for screening of a therapeutic agent by determining the alteration of the propensity of cells to interact in a population of cells due to addition of one or more test substance(s), wherein the population of cells comprises at least two distinguishable subpopulations of cells and wherein the method comprises the following steps:
(a) determination of the number of interactions between cells of a first distinguishable subpopulation and cells of a second distinguishable subpopulation;
(b) determination of the number of interactions in the cell sample by randomly assigning cells comprised in the cell sample to first and second distinguishable subpopulation of cells, wherein the absolute number of cells in the first and second distinguishable subpopulation of cells, respectively, are identical to the number of cells in the first and second distinguishable subpopulation of cells in (a);
(c) determination of the propensity of cells to interact by dividing (a) through (b), wherein propensity increases with the resulting number; and
(d) determination of the alteration of the propensity of cells to interact by comparing the propensity prior to addition of one or more test substance(s) and subsequent to addition of one or more test substance(s) and thereby determining whether the one or more test substance(s) qualify as therapeutic agent.
7 . (canceled)
8 . The method of claim 1 , wherein first and second distinguishable subpopulations are identical.
9 . The method of claim 1 , wherein step (b) is repeated and the determined number of interactions is averaged, preferably wherein step (b) is repeated more than at least 1000 times.
10 . The method of claim 1 , wherein the cells are (a) peripheral blood mononuclear cells (PBMCs) or (b) bone marrow cells.
11 . The method of claim 1 , wherein the disease is a myeloproliferative disorder, inflammatory disorder, latent virus infection, cellular growth disorder, cellular chemotaxis disorder, metabolic disorder or autoimmune disorder.
12 . The method of claim 10 , wherein the disease is leukemia or lymphoma.
13 . The method of claim 1 , wherein the cells are present in form of a monolayer.