Abstract: The present invention provides an epigenetic haemogram, also referred to as an epigenetic blood cell count that identifies the quantitative, comprehensive picture of cellular composition in a biological sample, wherein advantageously a normalization standard is used. The normalization standard is a nucleic acid molecule comprising at least one marker-region being specific for each of the blood cells to be detected, and at least one control-region being cell-unspecific, wherein said regions are present in the same number of copies on said molecule and/or a natural blood cell sample of known composition. Furthermore, the present invention relates to a kit and the use of a kit for performing the epigenetic assessment of comprehensive, quantitative cellular composition of a biological sample. The biological sample is derived from e.g.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying FoxP3-positive regulatory T cells, preferably CD25+CD4+ regulatory T cells of a mammal, comprising analysing the methylation status of at least one CpG position in the gene foxp3 or an orthologous or paralogous gene thereof, and the use of DNA-methylation analysis of the gene of the transcription factor FoxP3 for a detection and quality assurance and control of regulatory T cells. Furthermore, the present invention relates to a kit for performing the above methods as well as respective uses.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying CD8 positive subpopulations of a mammal, wherein said method comprises analyzing the bisulfite convertibility of at least one CpG position in the CD8 beta and CD8 alpha cell specific bisulfite convertibility gene region according to SEQ ID No. 1 and 2, wherein a bisulfite convertibility of at least one CpG position in said gene regions is indicative for a CD3+CD8+ and/or CD3+/?CD8+ cell. The analyses according to the invention can identify CD3+CD8+ and/or CD3+/?CD8+ cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood cells.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying certain immune cells of a mammal, comprising analysing the methylation status of at least one CpG position in the gene CCR6 and/or BLR1 or an orthologous or paralogous gene thereof, and the use of DNA-methylation analysis of the genes of the proteins CCR6 and/or BLR1 for a detection and quality assurance and control of certain immune cells. In particular, the present invention relates to analysing the methylation status of at least one CpG position in the gene CCR6 in T cells. Furthermore, the present invention relates to a kit for performing the above methods, as well as to respective uses.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying a subgroup of natural killer cells of a mammal, preferably CD3?, non T-lymphocyte derived NK cells, which often express the surface proteins CD56 and/or CD16, comprising analyzing the accessibility of the genomic DNA for OSBPL, such as OSBPL5, to bisulfite conversion and/or the methylation status of at least one CpG position in the genes for OSBPL, such as OSBPL5, in particular in their upstream and/or downstream regulatory regions, the promoter, introns, exons and introns exon borders and other conserved regions of said genes, wherein an increase of the accessibility of the genomic DNA and/or a demethylation in the sample as analyzed is indicative for said subgroup of NK cells. The analyses according to the invention can identify CD56+ cells and distinguish them from all other cells such as, for example, either CD56? and/or CD56 bright cells.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying CD8 positive subpopulations of a mammal, wherein said method comprises analyzing the bisulfite convertibility of at least one CpG position in the CD8 beta and CD8 alpha cell specific bisulfite convertibility gene region according to SEQ ID No. 1 and 2, wherein a bisulfite convertibility of at least one CpG position in said gene regions is indicative for a CD3+CD8+ and/or CD3+/?CD8+ cell. The analyzes according to the invention can identify CD3+CD8+ and/or CD3+/?CD8+ cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood cells.

Abstract: The present invention provides an epigenetic haemogram, also referred to as an epigenetic blood cell count that identifies the quantitative, comprehensive picture of cellular composition in a biological sample, wherein advantageously a normalization standard is used. The normalization standard is a nucleic acid molecule comprising at least one marker-region being specific for each of the blood cells to be detected, and at least one control-region being cell-unspecific, wherein said regions are present in the same number of copies on said molecule and/or a natural blood cell sample of known composition. Furthermore, the present invention relates to a kit and the use of a kit for performing the epigenetic assessment of comprehensive, quantitative cellular composition of a biological sample. The biological sample is derived from e.g.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying monocytes, comprising analyzing the methylation status of at least one CpG position in the mammalian gene region for parkin RBR E3 ubiquitin protein ligase (PARK2), wherein a de-methylation or lack of methylation of said gene region is indicative for a monocyte, when compared to a non-monocyte cell. The analyses according to the invention can identify monocytes on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The present invention furthermore provides an improved method for quantifying monocytes, in particular in complex samples. The method can be performed without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue. Also claimed are kits and specific oligonucleotides for use as primers or probes.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying B cells, comprising analyzing the methylation status of at least one CpG position in the mammalian gene region for Low density lipoprotein receptor-related protein 5 (LRP5), wherein a demethylation or lack of methylation of said gene region is indicative for a B cell, when compared to a non-B cell. The analyses according to the invention can identify B cells on an epi-genetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The present invention furthermore provides an improved method for quantifying B cells, in particular in complex samples. The method can be performed without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue. Further claimed are kits and specific primers and probes for identifying methylation.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying CD3CD4 positive T lymphocytes of a mammal, wherein the method comprises analyzing the bisulfite convertibility of at least one CpG position in the CD3+CD4+ T helper cell specific non-methylated bisulfite convertible region according to SEQ ID No. 1, wherein a bisulfite convertibility of at least one CpG position to at least 90%, preferably to at least 91% and more preferably to at least 92% and most preferred to at least 95% in the sample is indicative for a CD4+ T-lymphocyte cell, in particular a CD3+CD4+ T-lymphocyte cell. The present invention further relates to analyzing the bisulfite convertibility of at least one CpG position in the genes FLJ00060, FLJ38379, PPP6C, CD226, ZBTB7B and TNFAIP8 that are capable of positively identifying CD4 expressing cells in whole blood and segregate between CD4 and CD8 positive CD3 positive cells.

Abstract: The invention relates to a method for identifying a specific type and/or state of a mammalian cell in a sample, the method comprising a) analyzing the relative amount of accessible chromatin in regions that are specific for a cell-type and/or cellular state in the genome of the cell, b) comparing the relative amount of accessible chromatin in the regions with the relative amount of accessible chromatin in regions in the genome of the cell that are unspecific for the cell-type and/or cellular state, and c) deducing the specific type and/or state of said mammalian cell in the sample based on such comparison. The identifying further comprises a relative quantification of the specific cell type and/or state. The method can further comprise a diagnosis of a predisposition to a disease or a disease based on such identification. Kits and markers in regions of accessible chromatin are also described.

Abstract: The present invention relates to a method, in particular an in vitro method, for identifying certain immune cells of a mammal, comprising analyzing the methylation status of at least one CpG position in the gene CCR6 and/or BLR1 or an orthologous or paralogous gene thereof, and the use of DNA-methylation analysis of the genes of the proteins CCR6 and/or BLR1 for a detection and quality assurance and control of certain immune cells. In particular, the present invention relates to analyzing the methylation status of at least one CpG position in the gene CCR6 in T cells. Furthermore, the present invention relates to a kit for performing the above methods, as well as to respective uses.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying IL-17 expressing T cells in a blood and/or tissue sample derived from a mammal, comprising analysing the methylation status of at least one CpG position in the gene IL-17A, wherein a demethylation of said at least one CpG position in said sample when compared to an analogous position in a non IL-17 blood cell is indicative for a IL-17 positive CD4 positive T cell. The analyses according to the invention can identify IL-17 positive T cells and distinguish them from all other cells in complex samples, such as, for example, other blood cells. The present invention furthermore provides an improved method for quantifying IL-17 positive T cells in complex samples based on a comparison of the IL-17A methylation with a methylation of at least one marker selected from the group of CD3, FOXP3, and/or GAPDH.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying natural killer cells of a mammal, which often express the surface proteins CD 16 and/or CD56, comprising analyzing the methylation status of at least one CpG position in the CX3CR1 and/or FGR and/or NKG7 and/or GNLY genes, in particular their upstream regulatory regions, and in particular the promoter and other conserved regions of the genes CX3CR1 and/or FGR and/or NKG7 and/or GNLY, wherein a demethylation of at least one CpG in the analyzed sample to at least 70% is indicative for CD56 expressing NK cells, which might also be CD8+ or CD8?, CD56 dim or bright, CD 16+ or CD 16? NK cells. The methods of the present invention are useful for the detection and quality assurance and control of NK cells. Furthermore, the present invention relates to a kit for performing the above methods as well as respective uses of the inventive methods or kits.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying a subgroup of natural killer cells of a mammal, preferably CD3?, non T-lymphocyte derived NK cells, which often express the surface proteins CD56 and/or CD16, comprising analyzing the accessibility of the genomic DNA for OSBPL, such as OSBPL5, to bisulfite conversion and/or the methylation status of at least one CpG position in the genes for OSBPL, such as OSBPL5, in particular in their upstream and/or downstream regulatory regions, the promoter, introns, exons and introns exon borders and other conserved regions of said genes, wherein an increase of the accessibility of the genomic DNA and/or a demethylation in the sample as analyzed is indicative for said subgroup of NK cells. The analyses according to the invention can identify CD56+ cells and distinguish them from all other cells such as, for example, either CD56? and/or CD56 bright cells.

Abstract: The present invention relates to a method, in vitro or in vivo, for determining cancer patient survival, comprising analyzing the number and/or amount of tumor-infiltrating overall T-lymphocytes (oTLs) based on the methylation status of at least one CpG position in one or more of the genes for CD3 ?, -?, and -? in a tumor sample derived from said cancer patient, wherein a high number and/or amount of oTLs is indicative for a better survival of said cancer patient in a non-breast cancer, wherein in breast cancer a high number and/or amount of oTLs is indicative for an inferior survival of said patient. The present invention also relates to a respective kit for use in the methods of the invention.

Abstract: The present invention provides methods, nucleic acids and molecular markers for the characterization of cells, tissues and heterogeneous mixtures of cells. Specifically, it describes particular genes and genomic regions in which DNA methylation patterns are a consistent and characteristic property of different cell types, states and stages of differentiation. The invention is useful in determining the identity, composition, quality and potency of cells and cell populations. Furthermore, the invention will be useful in monitoring the differentiation of cells.

Abstract: The present invention relates to a method, in particular an in vitro method for identifying natural killer cells of a mammal, which often express the surface proteins CD 16 and/or CD56, comprising analysing the methylation status of at least one CpG position in the CX3CR1 and/or FGR and/or NKG7 and/or GNLY genes, in particular their upstream regulatory regions, and in particular the promoter and other conserved regions of the genes CX3CR1 and/or FGR and/or NKG7 and/or GNLY, wherein a demethylation of at least one CpG in the analyzed sample to at least 70% is indicative for CD56 expressing NK cells, which might also be CD8+ or CD8?, CD56 dim or bright, CD 16+ or CD 16? NK cells. The methods of the present invention are useful for the detection and quality assurance and control of NK cells. Furthermore, the present invention relates to a kit for performing the above methods as well as respective uses of the inventive methods or kits.