Methods and compositions related to joint inflammation diseases
Disclosed are compositions and methods related to joint inflammation diseases. Disclosed is the relationship between osteoclasts and inflammatory joint diseases and osteoclast precursor cells.
1. A method of diagnosing a subject with erosive arthritis comprising measuring how many osteoclast precursor cells (OCP) are in the blood of the subject and correlating the number of OCP in the blood of the subject with the presence of erosive arthritis, wherein a significant increase in the number of OCP in the subject relative to a healthy control subject having no erosive arthritis indicates the presence of erosive arthritis.
2. The method of claim 1 , wherein the OCP in the blood are obtained by collecting the subject's Peripheral Blood Mononuclear Cells (PBMCs).
3. The method of claim 2 , wherein the step of measuring comprises counting the number of cells comprising at least one marker selected from the group consisting of CD14, CD11a, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4 (CD49d), CD16, MHC Class II, B7.1, B7.2, CD40, and c-fms.
4. The method of claim 2 , wherein the step of measuring comprises counting the number of cells comprising at least two markers selected from the group consisting of CD14, CD11a, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4 (CD49d), CD16, MHC Class II, B7.1, B7.2, CD40, and c-fms.
5. The method of claim 2 , wherein the step of measuring comprises counting the number of cells comprising at least three markers selected from the group consisting of CD14, CD11a, CD11b, CD51/CD61, RANK, CCR1, CCR4,VCAM (CD106), VLA-4 (CD49d), CD16, MHC Class II, B7.1, B7.2, CD40, and c-fms.
6. The method of claim 2 , wherein the step of measuring comprises counting the number of cells comprising at least four markers selected from the group consisting of CD14, CD11a, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4 (CD49d), CD16, MHC Class II, B7.1, B7.2, CD40, and c-fms.
7. The method of claim 1 , wherein the amount of OCP is determined by staining the PBMC sample with fluorescently labeled antibodies for at least one marker selected from the group consisting of CD14, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4 (CD49d), CD16, CD 11a , MHC Class II, B7.1, B7.2, CD40, and c-fms and visualizing the cells with labeled antibody bound to at least one of CD14+, CD11b+, CD51/CD61+, RANK+, CCR1+, CCR4+, VCAM+(CD106), VLA-4+(CD49d), CD11a, MHC Class II, B7.1, B7.2, CD40, c-fms or CD16- (negative) using Fluorescence Activated Cell Sorting (FACS).
8. The method of claim 1 , wherein the number of OCP is further determined by removing a tissue sample from the subject and visualizing the sample using immunohistochemistry for at least one marker selected from the group consisting of CD14, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4(CD49d), CD 11a , MHC Class II, B7.1, B7.2, CD40, c-fms and CD16.
9. The method of claim 1 , wherein the amount of OCP is measured by obtaining a blood sample from the subject and staining the blood sample with Tartrate Resistant Acid Phosphatase (TRAP), counting how many multinucleated cells there are in the sample, and comparing the number of multinucleated cells in the blood sample from the subject to a number of multinucleated cells in a blood sample from a healthy control subject having no erosive arthritis, wherein a significant increase in the number of multinucleated cells in the sample from the subject than in the sample from the healthy control indicates erosive arthritis in the subject.
10. The method of claim 1 ,wherein the amount of OCP in the subject's blood is measured using FACS methods, Immunohistochemistry methods, Western methods, Southern methods, hybridization methods, reverse transcription polymerase chain reaction (RT-PCR) methods, enzyme-linked immunosorbent assay (ELISA) methods, enzyme-linked immunosorbent spot (ELISPOT) methods, microarray methods, bone wafer resorption methods or Immunoprecipitation methods.
11. The method of claim 1 , wherein measuring the number of OCP comprises identifying RANK, CD11b and CD14 positive cells in the blood sample.
12. The method of claim 1 , wherein the subject shows bone erosion on a radiograph.
13. The method of claim 1 , wherein the amount of OCP is measured by obtaining a blood sample from the subject and staining the blood sample with Tartrate Resistant Acid Phosphatase (TRAP), counting how many multinucleated cells there are in the sample from the subject, and comparing the number of multinucleated cells in the blood sample from the subject to a number of multinucleated cells in a blood sample from a healthy control subject having no erosive arthritis, wherein greater than 2.5 times more multinucleated cells in the sample from the subject than in the sample from the healthy control indicates erosive arthritis in the subject.
14. A method of diagnosing erosive arthritis comprising culturing peripheral blood mononuclear cells (PBMC) from a subject and assaying the number of osteoclasts formed and correlating the number of OCP in the PBMC of the subject with the presence of erosive arthritis, wherein a significant increase in the number of osteoclasts in the culture from the subject relative to the number of osteoclasts in a culture of PBMC from a healthy control subject without erosive arthritis indicates the subject has erosive arthritis.
15. The method of claim 14 , wherein assaying the number of osteoclasts formed comprises monitoring the amount of TRAP positive cells.
16. The method of claim 14 , wherein assaying the number of osteoclasts formed comprises monitoring the number of multinucleated cells.
17. The method of claim 14 , wherein the culture has no exogenous RANKL or M-CSF added.
18. The method of claim 14 , wherein addition of RANKL or M-CSF increases the number of osteoclasts in the culture from the subject relative to the number of osteoclasts in a culture of PBMC from a control subject without erosive arthritis, and wherein this increase indicates the subject has erosive arthritis.
19. A method of determining the presence of an erosive arthritis in a subject comprising, obtaining a PBMC sample from the blood of a subject, and measuring how many OCP are in the PBMC of the subject, wherein greater than 2.5 times more OCP in the PBMC of the subject than in a sample of PBMC from a control subject without erosive arthritis indicates the presence of erosive arthritis in the subject.
20. A method of determining whether a subject has erosive arthritis comprising isolating PBMC from the subject, and probing for the presence of three or more surface markers of mononuclear OCP, wherein the surface markers are selected from the group consisting of CD14, CD11b, CD51/CD61, RANK, CCR1, CCR4, VCAM (CD106), VLA-4 (CD49d), CD 11a , MHC Class II, B7.1, B7.2, CD40, c-fms and CD16, and wherein a significant increase in the number of mononuclear OCP in the PBMC of the subject as determined by the presence of the three or more surface markers relative to a healthy control without erosive arthritis indicates that the subject has erosive arthritis.
21. The method of claim 20 , wherein the surface markers are analyzed by FACS.
22. The method of claim 20 , wherein probing for a surface marker comprises assaying for at least four surface markers.