Detection Of Soluble HLA Class Ⅰ And Its Serial Monitoring For Renal Transplant Rejection And Other Clinical Significance

Objective:1. To find the best test condition of ELISA for sHLA-I and stablize the method;2. To apply the method of quantification of sHLA-I to survey its normal level in Guangdong people;3. To apply the method of quantification of sHLA-I to survey its various level in blood componets;4. To report the result of serial monitoring of sHLA-I by ELISA technique in 10 renal transplant recipients.Methods:Sandwich ELISA assay was used,W6/32 McAb serving as solid phase antibody and 3 2m antibody as the first antibody.The second antibody -HRP conjugate was added for coloration.Standard curve was obtained by sHLA-I standard reagent in serial dilution.The amount of sHLA-I in the samples was determined:1. The sHLA-I amount in sera of 60 Guangdong people was determined;2. The sHLA-I amount in blood componets of 20 donors was determined;3. The sHLA-I amount in sera of 10 renal transplant recipients wasdetermined serially. Results:1. Sensitivity of this assay is 2.84ng/ml.Coefficients of variation are 5.80 % within assays and 9.00 % between assays.The recovery rate is (98.57-100.15) %.The sHLA-I level of normal individual in Guangdong is (699.54 + 360.10)ng/ml.2. sHLA-I in red blood cells stored for 28 days and in random-donor platelets are significantly higher than that in other blood components and their amount is proportionate to the number of residual donor leukocytes as well as to the length of storage.3. It is found that sHLA-I level in sera of renal transplant recipients significantly elevates in acute rejection episodes and does not change significantly in stable condition.Immunosuppressive agents affect obviously the sHLA-I levels in treatment process.Conclusion:1. Sandwich ELISA assay is a sensitive,specific and stable technique.2. Blood components with different concentration of sHLA-I may be chosen for clinical transfusion,3. It is suggested that the monitoring of sHLA-I is very important to evaluate the effect of rejection episide for renal transplant recipients.