Method for estimation of the amount of specific cell types
The present invention relates to the estimation of the amount of subtypes of specific cells, for example the number of certain subtypes of leukocytes, by measurements of unique proteins in extracts of blood and other biological material. The knowledge of the number or amount of specific subtypes of white cells is important in the clinical diagnosis and surveillance of subjects with inflammatory disease including infectious disease, cancer, allergy/asthma etc.
1. A method for in vitro estimating an amount of specific cells in a patient sample using cell specific molecules, the method comprising
a) extracting cell specific molecules from cells contained in an aliquot of the patient sample;
b) measuring the concentration of at least two cell specific molecules in the extracted aliquot by immunological assay; and
c) estimating the amount of specific cells that contained the at least two cell-specific molecules from a ratio between the measured concentrations of the cell specific molecules.
2. The method according to claim 1 , wherein the extraction is conducted with cationic detergent.
3. The method according to claim 2 , wherein the detergent is CTAB (N-cetyl-N,N,N-trimethylammonium bromide).
4. The method according to claim 1 , wherein the patient sample is whole blood.
5. The method according to claim 1 , wherein the cell specific molecules are intracellular and extracellular proteins or cell surface markers.
6. The method according to claim 1 , wherein the concentration of each cell specific molecule is correlated with a number of a subtype of cells.
7. The method according to claim 1 , wherein the cell specific molecules comprise a neutrophil protein.
8. The method according to claim 7 , wherein the neutrophil protein comprises MPO (myeloperoxidase), HNL (human neutrophil lipocalin) or lactoferrin.
9. The method according to claim 1 , wherein the cell specific molecules comprise an eosinophil protein.
10. The method according to claim 9 , wherein the eosinophil protein comprises EPX (eosinophil protein x) or EPO (eosinophil peroxidase).
11. The method according to claim 1 , wherein the cell specific molecules comprise a basophil protein.
12. The method according to claim 11 , wherein the basophil protein is BB1.
13. The method according to claim 1 , wherein the cell specific molecules comprise a thrombocytic protein for measuring thrombocytes.
14. The method according to claim 13 , wherein the thrombocytic protein is β-thromboglobulin.
15. The method according to claim 1 , wherein the cell specific molecules comprise cell surface markers for measuring B-lymphocytes and CD3 T-lymphocytes.
16. The method according to claim 15 , wherein the cell surface markers are CD20.
17. The method according to claim 1 , wherein the cell specific molecules comprise cell surface markers for measuring different lymphocyte populations.
18. The method according to claim 17 , wherein the cell surface markers are CD4 or CD8.
19. The method according to claim 1 , wherein the cell specific molecules are CD14 or lysozyme for measuring monocytes.
20. The method according to claim 1 , wherein the cell specific molecules comprise myeloperoxidase and lactoeffin.