Immunological signatures and parameters predicting therapeutic responses to anti-TNF therapy
The invention relates to a method of predicting therapeutic responses to TNF blockers before anti-TNF therapy comprising analyzing immune parameters to selected stimuli in patients before therapy and its use for anti-TNF therapy. The invention relates also to a method of determining a predictive biomarker of response to anti-TNF therapy and to the use of the predictive biomarker obtained by the method.
1. A method, comprising:
a) obtaining a culture of immune cells from a biological sample taken from a patient having a chronic inflammatory disease, wherein the biological sample is collected prior to an anti-TNF therapy;
b) stimulating the culture of immune cells with an agent that stimulates the innate or adaptive immune system; and
c) measuring the expression level in the culture of immune cells of at least one cytokine and/or matrix metalloproteinase (MMP).
2. The method according to claim 1 , wherein said chronic inflammatory disease is rheumatoid arthritis or a disease of the spondyloarthritis (SpA) group.
3. The method of claim 2 , wherein the SpA group disease is selected from psoriatic arthritis, reactive arthritis, arthritis related to inflammatory bowel disease, a subgroup of juvenile chronic arthritis and ankylosing spondylitis.
4. The method according to claim 1 , wherein said agent that stimulates the innate or adaptive immune system is chosen from a heat-killed live, attenuated or killed microbe and a Toll-like receptor agonist.
5. The method of claim 1 , wherein the culture of immune cells is stimulated with Helicobacter pylori or Influenza A virus.
6. The method of claim 1 , wherein the cytokine is selected from the group consisting of: IFN-gamma, IL-1 beta, IL-12p70, IL-1ra, IL-17, IL-2, IL-4, MCP1, MIP-1 alpha, MIP-1 beta, TNF-alpha and TNF-beta and the MMP is MMP-9.
7. The method of claim 1 , wherein the expression level is measured by measuring mRNA expression level.
8. The method of claim 1 , wherein the expression level is measured by measuring protein expression level.
9. The method of claim 6 , wherein the expression level of the at least one of IL-1ra, MIP-1beta, and TNF-alpha is increased.
10. The method of claim 6 , wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased.
11. The method of claim 1 , wherein the biological sample taken from a patient is whole blood.
12. A method, comprising:
a) obtaining a first culture of immune cells from a biological sample taken from a patient having a chronic inflammatory disease, wherein the biological sample is collected prior to an anti-TNF therapy;
b) stimulating the first culture of immune cells with a first agent that stimulates the innate or adaptive immune system;
c) measuring the expression level in the first culture of immune cells of at least one cytokine and/or matrix metalloproteinase (MMP) in the first culture of immune cells;
d) obtaining a second culture of immune cells from the patient;
e) stimulating the second culture of immune cells with a second agent that stimulates the innate or adaptive immune system; and
f) measuring the expression level in the second culture of immune cells of at least one cytokine and/or matrix metalloproteinase (MMP) in the second culture.
13. The method according to claim 12 , wherein said chronic inflammatory disease is rheumatoid arthritis or a disease of the spondyloarthritis (SpA) group.
14. The method of claim 13 , wherein the SpA group disease is selected from psoriatic arthritis, reactive arthritis, arthritis related to inflammatory bowel disease, a subgroup of juvenile chronic arthritis and ankylosing spondylitis.
15. The method of claim 14 , wherein the SpA group disease is ankylosing spondylitis.
16. The method of claim 12 , wherein the expression level of IL-1ra, MIP-1beta, and TNF-alpha is measured in the first culture of immune cells and the expression level of IL-1ra, MIP-1beta, and TNF-alpha is measured in the second culture of immune cells.
17. The method of claim 12 , wherein the expression level is measured by measuring mRNA expression level.
18. The method of claim 12 , wherein the expression level is measured by measuring protein expression level.
19. The method of claim 12 , wherein the expression level of the at least one of IL-1ra, MIP-1beta, and TNF-alpha is increased in the first culture of immune cells and in the second culture of immune cells.
20. The method of claim 12 , wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased in the first culture of immune cells; and wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased in the second culture of immune cells.
21. A method, comprising:
a) obtaining a culture of immune cells from a biological sample taken from a patient having a chronic inflammatory disease, wherein the biological sample is collected prior to an anti-TNF therapy;
b) stimulating the culture of immune cells with an agent that stimulates the innate or adaptive immune system; and
c) measuring the expression level in the culture of immune cells of IL-1ra, MIP-1beta, and TNF-alpha;
wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased.
22. A method, comprising:
a) obtaining a first culture of immune cells from a biological sample taken from a patient having a chronic inflammatory disease, wherein the biological sample is collected prior to an anti-TNF therapy;
b) stimulating the first culture of immune cells with a first agent that stimulates the innate or adaptive immune system;
c) measuring the expression level in the first culture of immune cells of IL-1ra, MIP-1beta, and TNF-alpha;
d) obtaining a second culture of immune cells from the patient;
e) stimulating the second culture of immune cells with a second agent that stimulates the innate or adaptive immune system; and
f) measuring the expression level in the second culture of immune cells of IL-1ra, MIP-1beta, and TNF-alpha;
wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased in the first culture of immune cells; and
wherein the expression level of IL-1ra is increased, the expression level of MIP-1beta is increased, and the expression level of TNF-alpha is increased in the second culture of immune cells.