Method of determining chymase activity with secretory leukocyte protease inhibitor
It is now discovered that human chymase cleaves human SLPI at a specific site and that this cleavage can be used as an indicator of chymase activity. The present invention provides methods of diagnosing a chymase-associated disease or evaluating the efficiency of a treatment of a chymase-associated disease in a human subject by measuring SLPI processing, as well as other related methods and compositions.
1. A method of detecting a chymase-associated disease in a human subject, comprising the steps of:
a. obtaining a biological sample from the human subject;
b. measuring the level of human Secretory Leukocyte Protease Inhibitor (SLPI) cleavage by human chymase in the biological sample, wherein the level of human SLPI cleavage by human chymase is measured as a ratio of a human SLPI fragment resulting from chymase cleavage to the full length SLPI present in the biological sample; and
c. comparing the level measured from step b) to a control of the level of human SLPI cleavage by human chymase in a healthy human subject, wherein an elevated level of human SLPI cleavage by human chymase compared to said control indicates that the human subject has a chymase-associated disease or has increased risk of a chymase-associated disease.
2. The method of claim 1 , wherein the biological sample is saliva.
3. The method of claim 1 , wherein the chymase-associated disease is selected from the group consisting of asthma, allergic rhinitis, fibrosis, hypertension, cardiac hypertrophy, heart failure, rheumatoid arthritis, diabetic nephropathy, cystic fibrosis, COPD, and inflammatory diseases.
4. The method of claim 1 , wherein the full length human SLPI comprises the amino acid sequence of SEQ ID NO: 1.
5. The method of claim 1 , wherein the human SLPI fragment resulting from chymase cleavage consists essentially of the amino acid sequence of SEQ ID NO: 3.
6. The method of claim 1 , wherein the human SLPI fragment resulting from chymase cleavage consists essentially of the amino acid sequence of SEQ ID NO: 4.
7. The method of claim 1 , wherein the full length human SLPI consists essentially of the amino acid sequence of SEQ ID NO: 2.
8. A method of evaluating the effectiveness of a treatment to a chymase-associated disease in a human patient, comprising the steps of:
a. obtaining a biological sample from the human patient;
b. measuring the level of human SLPI cleavage by human chymase in the biological sample, wherein the level of human SLPI cleavage by human chymase is measured as a ratio of a human SLPI fragment resulting from chymase cleavage to the full length SLPI present in the biological sample; and
c. comparing the level measured from step b) to a control of the level of human SLPI cleavage by human chymase in the human patient prior to the treatment, wherein a decreased level of human SLPI cleavage by human chymase compared to said control indicates that the treatment to a chymase-associated disease in said patient is effective.
9. The method of claim 8 , wherein the biological sample is saliva.
10. The method of claim 8 , wherein the chymase-associated disease is selected from the group consisting of asthma, allergic rhinitis, fibrosis, hypertension, cardiac hypertrophy, heart failure, rheumatoid arthritis, diabetic nephropathy, cystic fibrosis, COPD, and inflammatory diseases.
11. The method of claim 8 , wherein the full length human SLPI consists essentially of the amino acid sequence of SEQ ID NO: 1.
12. The method of claim 8 , wherein the human SLPI fragment resulting from chymase cleavage consists essentially of the amino acid sequence of SEQ ID NO: 3.
13. The method of claim 8 , wherein the human SLPI fragment resulting from chymase cleavage consists essentially of the amino acid sequence of SEQ ID NO: 4.
14. The method of claim 8 , wherein the full length human SLPI consists essentially of the amino acid sequence of SEQ ID NO: 2.
15. The method of claim 8 , wherein the treatment to a chymase-associated disease in a human patient involves a compound that decreases the biological activity of chymase.