Diagnosis and monitoring of systemic lupus erythematosus and of scleroderma
View Patent ↗Methods for diagnosing and monitoring systemic lupus erythematosus (SLE) or scleroderma by determining, in a blood sample from the individual being diagnosed or monitored, complement component C4d deposited on surfaces of red blood cells in the sample, and optionally also determining complement receptor CR1 deposited on the red blood cell surfaces. For diagnosis this is compared with the quantity of C4d (and optionally CR1) present on red blood cells of normal individuals. For monitoring it is compared with a value in a sample or samples previously obtained from the individual patient. The comparison may be made with individual values for C4d and CR1 and/or with a ratio of the two found in normal individuals.
1. A method for diagnosing systemic lupus erythematosus in an individual, comprising:
(a) detecting, in a blood sample from the individual containing red blood cells, quantities of complement component C4d and complement receptor CR1 both of which are on surfaces of red blood cells in the sample,
(b) comparing said quantities with, respectively, the quantities of component C4d and receptor CR1 both on surfaces of red blood cells in samples from individuals not having systemic lupus erythematosus, and
(c) diagnosing systemic lupus erythematosus in the individual by observing a significantly increased quantity in complement component C4d on surfaces of red blood cells and a significantly decreased quantity of receptor CR1 on surfaces of red blood cells in said sample from the individual compared to said samples from said individuals not having systemic lupus erythematosus.
2. The method according to claim 1 in which step (b) comprises comparing the ratio of C4d:CR1 on surfaces of red blood cells in the sample with the ratio of C4d:CR1 on surfaces of red blood cells of individuals not having systemic lupus erythematosus, wherein a significantly higher ratio of C4d:CR1 on surfaces of red blood cells in the sample with the ratio of C4d:CR1 on surfaces of red blood cells of individuals not having systemic lupus erythematosus is indicative of a diagnosis of systemic lupus erythematosus.
3. The method according to claim 1 , in which the detection of C4d and CR1 are conducted by a method comprising binding the C4d to a conjugate of a monoclonal antibody specific for component C4d with a first labeled moiety, binding the CR1 to a conjugate of a monoclonal antibody specific for CR1 with a second labeled moiety, and determining the first and second labeled moieties.
4. The method according to claim 3 , in which the labeled moieties are fluorescent moieties.
5. The method according to claim 4 in which the fluorescent moieties are determined by determining the mean fluorescence channel using flow cytometric analysis.
6. A method of monitoring disease activity of systemic lupus erythematosus in an individual, comprising:
(a) detecting, in a blood sample from the individual containing red blood cells, quantities of complement component C4d and complement receptor CR1 both of which are on surfaces of the red blood cells in the sample,
(b) comparing said quantities with, respectively, the quantities of component C4d and receptor CR1 both on surfaces of red blood cells in a sample previously obtained from the individual, and
(c) monitoring disease activity of systemic lupus erythematosus in the individual by observing a change in said quantities of complement component C4d on surfaces of red blood cells and complement receptor CR1 on surfaces of the red blood cells in said sample compared to said sample previously obtained from the individual, wherein systemic lupus erythematosus disease activity in said individual is indicated by a significantly increased quantity in complement component C4d on surfaces of red blood cells and a significantly decreased quantity of complement receptor CR1 on surfaces of red blood cells in said sample from the individual compared to said sample previously obtained from the individual.
7. The method according to claim 6 , in which step (b) comprises comparing the ratio of C4d:CR1 on surfaces of red blood cells in the sample with the ratio of C4d:CR1 on surfaces of red blood cells previously obtained from the individual, wherein a significantly higher ratio of C4d:CR1 on surfaces of red blood cells in the sample compared to the sample previously obtained from the individual is indicative of systemic lupus erythematosus disease activity.
8. The method according to claim 6 , in which the detection of C4d and CR1 is conducted by a method comprising binding the C4d to a conjugate of a monoclonal antibody specific for C4d with a first labeled moiety, binding the CR1 to a conjugate of a monoclonal antibody specific for CR1 with a second labeled moiety, and determining the first and second labeled moieties.
9. The method according to claim 8 , in which the labeled moieties are fluorescent moieties.
10. The method according to claim 9 in which the fluorescent moieties are determined by determining the mean fluorescence channel using flow cytometric analysis.
11. A method of diagnosing systemic lupus erythematosus in an individual, comprising:
(a) detecting, in a blood sample from the individual containing red blood cells, a quantity of complement component C4d on surfaces of red blood cells in the sample,
(b) comparing said quantity with the quantity of component C4d on surfaces of red blood cells in samples from individuals not having systemic lupus erythematosus, and
(c) diagnosing systemic lupus erythematosus in the individual by observing a significantly increased quantity in complement component C4d on surfaces of red blood cells in said sample from the individual compared to said samples from individuals not having systemic lupus erythematosus.
12. The method according to claim 11 , in which the detection of C4d is conducted by a method comprising binding the C4d to a conjugate of a monoclonal antibody specific for C4d with a labeled moiety, and determining the labeled moiety.
13. The method according to claim 12 , in which the labeled moiety is a fluorescent moiety.
14. The method according to claim 13 , in which the fluorescent moiety is determined by determining the mean fluorescence channel using flow cytometric analysis.
15. A method of monitoring disease activity of systemic lupus erythematosus in an individual, comprising:
(a) detecting, in a blood sample from the individual containing red blood cells, a quantity of complement component C4d on surfaces of the red blood cells in the sample,
(b) comparing said quantity with the quantity of component C4d on surfaces of red blood cells in a sample previously obtained from the individual, and
(c) monitoring disease activity of systemic lupus erythematosus in the individual by observing a change in the quantity of complement component C4d on surfaces of the red blood cells in said sample, wherein systemic lupus erythematosus disease activity in said individual is indicated by a significantly increased quantity in complement component C4d on surfaces of red blood cells in said sample from the individual compared to said sample previously obtained from the individual.
16. The method according to claim 15 , in which the detection of C4d is conducted by a method comprising binding the C4d to a conjugate of a monoclonal antibody specific for C4d with a labeled moiety, and determining the first labeled moiety.
17. The method according to claim 16 , in which the labeled moiety is a fluorescent moiety.
18. The method according to claim 17 in which the fluorescent moiety is determined by determining the mean fluorescence channel using flow cytometric analysis.
19. A method for diagnosing or monitoring systemic lupus erythematosus in an individual, comprising:
(a) automatically detecting, in a blood sample from the individual containing red blood cells, quantities of complement component C4d and complement receptor CR1, both of which are on surfaces of the red blood cells in the sample,
(b) automatically comparing said quantities with reference values for component C4d and receptor CR1, respectively, both on surfaces of red blood cells, and
(c) diagnosing systemic lupus erythematosus in the individual by observing a significantly increased quantity in complement component C4d on surfaces of red blood cells and a significantly decreased quantity of receptor CR1 on surfaces of red blood cells in said sample from the individual compared to said reference values; or
(d) monitoring disease activity of systemic lupus erythematosus in the individual by observing a change in said quantities of complement component C4d on surfaces of red blood cells and complement receptor CR1 on surfaces of the red blood cells in said sample compared to said reference values, wherein systemic lupus erythematosus disease activity in said individual is indicated by a significantly increased quantity in complement component C4d on surfaces of red blood cells and a significantly decreased quantity of complement receptor CR1 on surfaces of red blood cells in said sample from the individual compared to said reference values.
20. The method according to claim 19 , in which the reference values comprise a ratio of C4d:CR1, and wherein a significantly higher ratio of C4d:CR1 on surfaces of red blood cells in the sample from the individual compared to the reference value ratio of C4d:CR1 is indicative of a diagnosis of systemic lupus erythematosus or is indicative of systemic lupus erythematosus disease activity in an individual with systemic lupus erythematosus.
21. A method for diagnosing or monitoring systemic lupus erythematosus in an individual, comprising:
(a) automatically detecting, in a blood sample from the individual containing red blood cells, a quantity of complement component C4d on surfaces of the red blood cells in the sample,
(b) automatically comparing said quantity with a reference value for component C4d on surfaces of red blood cells, and
(c) diagnosing systemic lupus erythematosus in the individual by observing a significantly increased quantity in complement component C4d on surfaces of red blood cells in said sample from the individual compared to said reference values; or
(d) monitoring disease activity of systemic lupus erythematosus in the individual by observing a change in the quantity of complement component C4d on surfaces of the red blood cells in said sample compared to said reference values, wherein systemic lupus erythematosus disease activity in the individual is indicated by a significantly increased quantity in complement component C4d on surfaces of red blood cells in said sample from the individual compared to said reference values.