Study On Prognostic Impact Of Pre-transplant Serum IP-10, Mig And BLC Levels And CXCR3 Polymorphisms On Renal Transplantation And Diagnostic Value Of Urinary IP-10,Mig And MIP-3α Levels For Early Renal Allograft Dysfunction

Objective. To investigate the influence of pre-transplant serum IP-10, Mig and BLC levels on acute rejection episodes and renal allograft survival, find non-invasive and practical laboratory tools for evaluating immune state, predicting acute rejection episodes and renal graft failure and tailoring immunosuppressive therapy.Methods. A retrospective cohort of 149 patients undergoing renal transplants ( RTX ) between Aug. 2001 and Dec. 2005, 92 males and 57 females, with their blood samples preserved was studied. 69 of 149 recipients had undergone biopsy-proved rejection within first 6 months after RTX; the others, with stable allograft function as RTX status at protocol allograft biopsy, had no acute rejection episode. Clinical and epidemiological data were investigated, retrospectively. The pre-transplant serum levels of 1P-10, Mig and BLC were determined by ELISA.determined by ELISA.Results. Pre-transplant serum Mig and BLC levels in RTX recipients were significantly higher than in healthy control subjects. Pre-transplant serum IP-10 and BLC levels in recipients undergoing rejection episodes were higher than in non-rejection group. The study showed an increased rate of rejection episode in recipients with serum IP-10 levels above 3.57 pg/ml in comparison with below 3.57 pg/ml (59.74% vs 21.95%;X~2=17.64, P=0.001) . Pre-transplant serum IP-10 levels and age were independent risk factors of acute rejection within first 6 months after RTX ( relative risk: 1.565, 0.893; 95% CI: 0.888-2.924, 0.814-0.980). Acute rejection episodes, pre-transplant serum IP-10 and BLC levels were risk factors of renal allograft failure ( P=0.000; P=0.034, P= 0.001, respectively). There were no significant differences in age, sex, times in dialysis, type of dialysis, primary disease, cold ischemia time, warm ischemia time, PRA, HLA mismatch, and preoperative serum Mig levels between recipients who lost the graft and those who did not. When life-time analysis was performed after the assignment of all patients to four groups according to A, B, C, and D group based on serum BLC levels , the 5-year survival rates of grafts were 97.4%, 92.8%, 80.3% and 60.3%, respectively( A(?) 89.69 pg/ml, n=37; B(?)120.19 pg/ml, n=37; C(?) 172.33 pg/ml, n=38; D>172.33 pg/ml, n=37; P=0.0005 ). The effect of serum BLC levels on graft survival rates appeared to be mainly operating within the first year after RTX, be maitained at 5 years' follow-up.Conclusions. Pre-transplant serum IP-10 levels and age are independent risk factors to predict acute rejection within first 6 months after RTX. Pre-transplant serum IP-10 and BLC levels and rejection episodes were risk factors of renal allograft failure.Part 2. Diagnostic value of urinary IP-10 Mig and MIP-3α levels for early renal allograft dysfunctionObjective. To investigate the relationship between early renal allograft dysfunction and urinary IP-10, Mig, and MIP-3α levels, explore the diagnostic and predictive value of urinary IP-10, Mig, and MIP-3α levels as indicators to diagnose early renal allograft dysfunctionMethods. A retrospective cohort of 222 patients undergoing RTX and renal allograft biopsy between Aug. 2001 and Dec. 2005, with their urine samples preserved was studied. 67 of the 222 biopsies within first 6 months after RTX were classified as acute rejection, 119 with stable allograft function as RTX status, 13 as subclinical acute rejection (SAR), 11 as acute tubular necrosis (ATN), and 12 with renal allograft dysfunction as biopsy-proved RTX status were classified as presumed acute rejection. Urine specimens were collected at renal biopsy, 2 weeks, 4 weeks, 6 weeks and 8 weeks after RTX, respectively. Clinical and epidemiological data were investigated retrospectively. The urinary IP-10, Mig and MIP-3α levels were determined by ELISA. Results. There were no differences between urinary IP-10 levels of healthy control subjects, 2 weeks, 4 weeks, 6 weeks, and 8 weeks after RTX ( 3.72± 2.32, 2.04 ± 1.16, 2.95±2.94, 0.66 ± 0.46, and 0.44 ± 0.18 pg/μmol Cr, respectively; P=0.33 ). There were no differences between urinary Mig levels of 2, 4, 6, 8 weeks after RTX ( 34.82 ±8.76, 25.66±7.61, 20.78 ± 8..90, 25.16 ± 9.54 pg/μmol Cr, respectively; P=0.725 ), which were higher than those of healthy control subjects ( 13.77 ± 1.91 pg/μmol Cr, P<0.001). Urinary MIP-3αlevels of 2 and 4 weeks after RTX were higher than those of 6 and 8 weeks after RTX and healthy control subjects (6.08 ± 1.57, 4.13 ± 1.80, 0.46±0.29, 0.83 ± 0.48, 0.82 ± 0.19 pg/μmol, respectively ). Urinary IP-10, Mig and MIP-3α levels at the time of graft biopsy were significantly elevated in RTX recipients with acute rejection and ATN. Urinary IP-10 and MEP-3α levels in recipients with ATN were significantly higher than in recipients with acute rejection. Urinary IP-10, Mig and MIP-3α levels in recipients as RTX status at time of protocol-biopsy were lower than in other groups. Receiver-operating characteristic curve characteristic (ROC) analysis demonstrated that early acute renal allograft dysfunction can be predicted with a sensitivity of 58% and a specificity of 92.9% using the cutoff value of 5.74 pg/μmol Cr of IP-10, with a sensitivity of 76.7% and a specificity of 76.6% using the cutoff value of 25.52 pg/μmol Cr of Mig, and with a sensitivity of 55.8% and a specificity of 83.9% using the cutoff value of 0.84 pg/μmol Cr of MIP-3α (P <0. 0001; P <0. 0001; P <0. 0001). The calculated area under the curve was 0.742 ( 95% CI 0.659 to 0.824 ) for urinary IP-10 levels, 0.805 (95% CI 0.733 to 0.876) for urinary Mig levels, 0.668 (95% CI 0.578 to 0.757) for urinary MIP-3α levels. 9 of 11 recipients with ATN had elevated urinary levels of all of three indicators, 2 had elevated level of two of all three. Except that 5 had no elevated levels of that three, 25 of 67 recipients with acute rejection had elevated urinary levels of all of that three; 37 had elevated urinary levels of one or two of that three. 12 of 13 with SAR had elevated urinary levels of one or two of three. All of 12 with presumed acute rejection had elevated urinary levels of one or two of that three. 84 of 119 with RTX status at protocol allograft biopsy had no elevated urinary levels of that three; 34 had elevated levels of one or two of that three; 1 with urinary infection, had elevated levels of all of that three. Except 4 with vascularrejection, 45 recipients with acute rejection episode within first 2 months after RTX reached a level less than the cutoff of that three within 1 to 4 weeks after anti-rejection therapy. Except that 1 as biopsy-proved renal cortical necrosis, the recipients with ATN did that within 1 to 4 weeks after RTX. Immunohistologic analysis of allograft biopsies showed exuberant expression of MIP-3α in tubular epithelial cells during acute rejection and ATN whereas it was absent in grafts with stable function as RTX status at the time of allograft protocol-biopsies.Conclusions. Urinary IP-10, Mig and MIP-3α levels were indicators to diagnose renal allograft dysfunction, not useful for differentiating acute rejection from ATN. Combined with renal allograft biopsy, detection of urinary IP-10, Mig and MIP-3α levels will help to diagnose and differentiate acute rejection, ATN and SAR. Urinary IP-10, Mig and MIP-3α levels tended to decline with the improvement of renal allograft function. It remained to be studied further whether urinary IP-10, Mig and MIP-3α levels were indicators to predict acute rejection and effect of anti-rejection therapy.Part 3. The association between single nucleotide polymorphisms in CXCR3 and acute rejection in renal transplantationObjective. To study the association between CXCR3 gene single nucleotide polymorphisms ( SNPs) and acute rejetion in RTX.Methods. A retrospective cohort of 223 patients receiving their first cadaver RTX between Aug. 2001 and May 2006, with their blood samples preservedwas studied. Clinical and pathological data were investigated retrospectively. We genotyped the CXCR3 gene SNPs (Intron 1, c.12+234G>A; dbSNP: , rs2280964) using TaqMan SNP Genotyping Assays System by Taqman real-time PCR. The associations between CXCR3 gene SNPs and acute rejetion episode within the first 6 months after RTX were analyzed. Results Eighty-eight RTX recipients experienced acute rejecton within first 6 months after RTX. The CXCR3 gene SNPs were not significantly related to acute rejection episode within first 6 months after RTX ( R×C X~2 test, Male:P=0.36; Female: P=0.98).Conclusion Our data indicate that CXCR3-rs 2280964 gene SNPs had no association with the occurrence of acute rejection within first 6 months after RTX...