Detection of plasma cells,C4d deposits and donor-specific antibodies on sequential graft biopsies of renal transplant recipients with chronic dysfunction

BackgroundIn order to look for a relationship between humoral mechanisms of rejection and chronic allograft dysfunction, plasma cells, C4d deposits and donor-specific antibodies (DSA) were simultaneously sought on serial biopsies of kidney allograft recipients.Patients and methodsTen recipients with chronic dysfunction (G1) and 8 recipients with long-term normal graft function (G2) were included. Biopsies and serums were sampled at early graft dysfunction (T1), between 8 months and 2 years (T2) and after the third year following transplantation (T3).ResultsIn G1, plasma cells represented 12.3% (T1), 8.2% (T2) and 14.1% (T3) of mononuclear cells. The mean percentage of plasma cells was 11.6% in G1 versus 0.4% in G2 (p < 0.05). A progressive rise in C4d deposits was seen in G1, from 25% at T1 to 80% at T3. Donor-specific antibodies were identified in at least one serum sample of 60% of the patients in G1 and 12.5% of the patients in G2 (p = 0.012), whereas donor-specific antibodies were eluted from at least one biopsy of 50% of the patients in G1 and 12.5% of the patients in G2 (p = 0.03). In G1, C4d deposits were significantly associated with plasma cells (p = 0.0012) and anti-HLA Abs in serum samples and/or eluates (p = 0.026).ConclusionThis study shows that plasma cells, DSA and C4d are associated in renal transplants developing chronic rejection.     

Statut d’hydratation extracellulaire et natriurèse résiduelle en hémodialyse chronique : une étude transversale multicentrique

BackgroundIn dialysis patients, a misevaluation of dry weight may lead to an increased morbidity and mortality. The aim of this cross-sectional multicenter study was to evaluate the association between residual urinary sodium excretion and extracellular volume status in chronically treated hemodialysis patients.Patients and methodsDry weight was determined clinically and by whole-body bioimpedance spectroscopy (Body Composition Monitor, Fresenius Medical Care) prior to a mid-week session in 40 chronic hemodialysis patients with significant residual diuresis (more than 250 mL per day) and receiving treatment in four dialysis centers. Regarding their hydration status assessed by the Body Composition Monitor and in comparison to a healthy reference population, patients were assigned to 1 of the 3 categories: overhydrated, normohydrated and dehydrated. Urine output, urinary sodium excretion and residual renal function were measured for all patients within 30 days before dry weight assessment.ResultsThe median post-HD session FO was of −0.40 L (IQR: from −1.95 to +0.90) and the median residual urinary sodium excretion was of 64 mmol/L (IQR: 46–79). Among these patients, 16 were normohydated, 16 were dehydrated and 8 were overhydrated. There was a linear relationship between the hydration status after HD session and the urinary sodium excretion (estimate: 5.6 ± 1.5; p < 0.001). Compared with normohydrated patients, overhydrated patients had a higher residual urinary sodium excretion (estimate: 26 ± 10; p < 0.01).ConclusionIn this study, urinary sodium excretion is associated with the hydration status evaluated by whole-body bioimpedance spectroscopy.     

Increased levels of CD4+ and CD8+ T cells expressing CCR1 in patients developing allograft dysfunction; a cohort study

BackgroundLeukocyte infiltration into the graft has pivotal effects on kidney transplantation outcome. The present study sought to determine whether the expression of sequential chemokine receptors on CD4⁺ and CD8⁺ T cells in human renal allograft can predict clinical episodes.MethodsBlood samples from 52 consecutive renal transplant patients were evaluated at the time of transplantation and at three times (2, 90 and 180 days) after transplantation to analyze the expression of CCR1 and CXCR3 on CD4⁺ and CD8⁺ T cells by flowcytometry. A total of 30 biopsies, including protocol biopsy (n = 24) and cause biopsy (n = 6), were investigated according to the Banff criteria.ResultsThe mean percentage of CD4⁺ and CD8⁺ T cells expressing CCR1 was significantly increased in patients with allograft dysfunction (n = 25) (p = 0.006, p = 0.004). The mean fluorescence intensity of CXCR3 on CD4⁺ and CD8⁺ T cells were found to be significantly higher in graft dysfunction than that in well-functioning grafts. (p < 0.001, p = 0.007). Receiver Operating Characteristic (ROC) Curve Analysis showed that the calculated AUC was 0.86 at the third month for CD4⁺ CCR1⁺ and CD8⁺ CCR1⁺ (p < 0.001). Multiple logistic regression analysis showed that an increase in CD4⁺ expressing CXCR3 leads to a lower risk of graft dysfunction (OR = 0.37), while an increase in CD8⁺ expressing CCR1 results in a higher risk of graft dysfunction (OR = 3.66).ConclusionDuring renal transplantation, CD4⁺ and CD8⁺ T cells expressing CCR1 were increased in patients who developed graft dysfunction. These findings may prospectively predict allograft dysfunction, and help elucidate the underlying pathogenic mechanisms.     

Crioablación total o hemiglandular para el cáncer de próstata primario localizado: resultados oncológicos y funcionales a corto y medio plazo

IntroductionTo compare oncological, functional and post-operative outcomes of hemi (HC) vs. whole gland (WGC) cryoablation as first line treatment of localized prostate cancer.Material and methodSixty-six consecutive patients undertaking whole-gland cryoablation (WGC = 40) or hemi-cryoablation (HC = 26) in a tertiary referral centre between 2010 and 2018 were included. All patients had a low-intermediate risk prostate cancer according to D’Amico risk classification. Hemi-ablation was proposed in case of biopsy and prostate MRI proven unilateral prostate cancer. Primary endpoint was Cryotherapy Failure for which 3 definitions were considered and compared: 1) biochemical failure (> PSA nadir + ≥ 2 ng/mL), 2) positive prostate re-biopsy with Gleason score ≥ 7, 3) initiation of further prostate cancer treatment.ResultsMedian patients age at treatment was 74 [42-81] vs. 76 [71-80] years in WGC vs. HC group, respectively (p = .08). Low and intermediate D’Amico risk group were 15% and 85% vs. 23% and 77% (p = .75), respectively. Median follow- up time was 41 [1.5-99.0] vs. 27 [0.9-93] months (p = .03). Four-years cryotherapy failure free survival in WGC vs. HC were 69% vs. 53% with definition 1 (p = .24), 82% vs. 80% with definition 2 (p = .95), 83% vs. 77% with definition 3 (p = .73).Early and 1-year urinary continence were 60% and 83% in WGC vs. 72% and 83% in HC (p = .26). De novo impotency after cryotherapy was 75% vs. 46% (p = .33) in WGC vs. HC.ConclusionsIn our cohort of highly selected patients with unilateral low/intermediate risk PCa, hemi-cryoablation may provide similar oncological outcomes and less early complications compared to whole-gland cryoablation.     

Association between the CX3CR1 gene V249I polymorphism and delayed kidney allograft function

BackgroundFractalkine is a member of the chemokine family that acts as an adhesion molecule and as an extracellular chemoattractant promoting cellular migration. In this study, we analysed the association between the CX3CR1 gene V249I (rs3732379) SNP and renal allograft function.MethodsThe study enrolled 270 Caucasian kidney allograft recipients. The following parameters were recorded in each case: the recipient's age and gender, delayed graft function (DGF) defined as the need for dialysis in the first 7 days after transplantation, occurrence and number of episodes of acute rejection (AR), and chronic allograft dysfunction (CAD).ResultsDelayed graft function was diagnosed in 39.2% of individuals with the CC genotype, 22.7% with CT and 23.5% of those with the TT genotype. The differences were statistically significant (CC vs. TT + CT: OR = 2.17; 95% CI = 1.28–3.70, p = 0.0042). In multivariate analysis the CC genotype was an independent and significant predictor of higher risk of DGF. The distribution of genotypes and alleles of the CX3CR1 gene polymorphism among patients with and without AR as well as CAD did not differ significantly.ConclusionsThe results of this study suggest that the CX3CR1 gene V249I (rs3732379) SNP CC genotype is associated with increased risk of DGF.     

Study of the cytokine polymorphisms in correlation to rejection and graft survival in renal allograft donors and recipients from a homogenous Saudi population

ObjectivesAllograft outcome can be improved with the discovery of risk factors that influence adverse events and may allow individualization of patients' treatment. Rejection is the main hurdle to successful transplantation and the immune response is the key effecter to rejection development. Hence, the major objective of the present study was to assess the relationship between single nucleotide polymorphisms (SNPs) in 5 cytokine genes, HLA mismatch and graft outcome in a cohort of 100 Saudi kidney transplant recipients and 100 living related donors at a single transplant center.Materials & methodsGenotyping of the following positions: TNFA (? 308 G/A), TGFB1 (codon 10 T/C, codon 25 C/G), IL-10 (? 1082 G/A, ? 819 C/T, ? 592 C/A), IL-6 (? 174 C/G), and IFNG (+ 874 T/A) were performed.ResultsThe majority of the donors whose recipients presented with either cellular or antibody mediated graft rejection (90% and 100%) respectively were found to be significantly (p = 0.0351) associated with intermediate or high IL-10 producing haplotypes, compared to those with stable grafts (58.66%). Haplotypes linked with lower IL-10 production were not detected in the donors or their recipients with antibody mediated graft rejection compared to donors with stable graft (41.33%). The distribution of donor IL-10-1082 haplotypes (GG, GA, AA) showed a statistically significant association of IL-10-1082 GA genotype (p = 0.0351) with rejection, when grouped according to patients' rejection status. No other statistically significant deviations were observed in the donors' genotypes. Analyses of cytokine polymorphisms in the recipients revealed no significant association. Multivariate logistic regression analyses showed that only HLA-DRB1 mismatch significantly influenced graft loss (p = 0.0135).ConclusionThis study demonstrates that the donor IL-10 genotypes and HLA-DRB1 mismatch are key determinants in graft outcome after renal transplantation.     

Deleterious impact of C3d-binding donor-specific anti-HLA antibodies after pediatric liver transplantation

BackgroundThe prevalence and clinical impact of anti-HLA donor-specific antibodies (DSA) after liver transplantation (LT) have not been extensively studied, especially in pediatric population.MethodsThe present cross-sectional study included 100 patients who underwent a first LT in childhood. Anti HLA immunization study was performed at a single time point during routine follow-up using Luminex® single antigen tests with classical anti-IgG conjugate and anti-C3d conjugate.ResultsThe main indication for LT was biliary atresia (52%) and median age at LT was 4.6 years. The median time between LT and DSA assessment was 7.8 years (range 1–21 years). DSA was identified in twenty-four patients (24%) after LT, with a prevalence of 8%, 28%, 33%, 50%, respectively 0–5 years, 5–10 years, 10–15 years and > 15 years after LT. DSA were mainly class II (23/24) with a mean MFI of 9.731 ± 5.489 and 18 (79.3%) were C3d-binding DSA. Multivariate analysis disclosed that time elapsed since LT (p < 0.01) and history of fulminant hepatitis (p = 0.04) were significantly associated with a higher rate of DSA. Liver function tests (at time of DSA assessment) were not different according to the presence or not of DSA (or C3d-binding DSA). Regarding histology, the DSA group had a higher rate of chronic rejection, cirrhosis and centrilobular fibrosis or cirrhosis. In addition, patients with C3d-binding DSA and high MFI (> 10,000) had a significant poorer long-term graft survival (p = 0.03).ConclusionIn our pediatric cohort of LT, prevalence of DSA was high and increased regularly with time. Presence of C3d positive-DSA with high MFI was associated with a higher rate of graft loss.     

Analysis of parathyroid graft rejection suggests alloantigen-specific production of nitric oxide by iNOS-positive intragraft macrophages

BackgroundDuring acute rejection of organ or tissue allografts T cells and macrophages are dominant infiltrating cells. CD4-positive T cells are important for the induction of allograft rejection and macrophages are important effector cells mediating cytotoxicity via production of nitric oxide (NO) by the inducible NO-synthase (iNOS). In the present study we analysed whether the destruction of primarily nonvascularised parathyroid allografts is also mediated by iNOS-positive macrophages.MethodsHypocalcaemic Lewis rats received parathyroid isografts (from Lewis donors) and allografts (from Wistar Furth donors), respectively, under the kidney capsule. Levels of serum calcium above 2 mmol/L correlated with normal parathyroid function and below 2 mmol/L with parathyroid rejection. Accelerated parathyroid allograft rejection was induced by immunisation of Lewis recipients with the allogeneic peptide P1.ResultsDetermination of serum calcium levels is a useful parameter to control parathyroid graft function, and therefore to determine allograft rejection. Macrophages positive for both major histocompatibility complex (MHC) class II molecules and costimulatory molecules accumulated in iso- and allografts, but iNOS-positive macrophages were only detectable in allografts in the presence of activated CD4-positive T cells. These results confirm a cooperation between activated T cells and intragraft macrophages to induce macrophage iNOS expression. Recipients immunised with the allogeneic peptide P1 demonstrated accelerated rejection of allografts (mean ± SD: 9.2 ± 0.9 days) in contrast to nonimmunised animals (mean ± SD: 15.8 ± 1.8 days). Allografts of P1-immunised animals were infiltrated faster by activated CD4-positve T cells and, in addition, the infiltrates of iNOS-positive macrophages were stronger than those in allografts of nonimmunised animals.ConclusionsIntragraft iNOS-positive macrophages seem to be able to produce cytotoxic NO involved in the killing of allogeneic cells during the alloimmune response against primarily nonvascularised parathyroid organ grafts. Infiltrates of iNOS-negative macrophages found in parathyroid isografts were caused by antigen-independent inflammation triggered by surgically induced injury. The absence of activated T cells in isografts and their presence in allografts underlines their importance in inducing macrophage iNOS expression.     

Clinical relevance of pretransplant anti-HLA donor-specific antibodies: Does C1q-fixation matter?

Anti-HLA donor-specific antibodies (DSA) identified by single antigen bead array (SAB) are questioned for their excess in sensitivity and lack of event prediction after transplantation.Population and methodsWe retrospectively evaluated specific types of preformed DSA (class I, class II or C1q-fixing) and their impact on graft survival. Kidney transplantations performed across negative CDC-crossmatch were included (n = 355). Anti-HLA antibodies were tested using SAB to identify DSA and their capacity to fix C1q.ResultsTwenty-eight patients with pretransplant DSA⁺ with MFI > 2000 were selected to assess C1q fixation. DSA were C1q + in 15 patients and C1q- in 13, without significant differences in demographics, acute rejection, graft loss or renal function. The maximum MFI of DSA in patients with C1q-fixing DSA was significantly higher (p = 0.008). Patients with DSA class-I suffered more antibody-mediated rejection (AMR) and had worse graft survival than class-II. The capacity of DSA I to fix C1q did not correlate with rejection, graft function or graft loss.ConclusionsC1q testing in pretransplant sera with DSA was unable to predict acute antibody-mediated rejection or early graft loss, but the presence of DSA class I compared to DSA only class II did. Despite non-fixing complement in vitro, pretransplant C1q-negative DSA I can mediate rejection and graft loss.     

Increased alloreactivity and adverse outcomes in obese kidney transplant recipients are limited to those with diabetes mellitus

Previous studies on patient and allograft outcomes of obese kidney transplant recipients (KTRs) remain controversial. To what extent obesity-related comorbidities contribute to adverse outcomes, however, hasn't been addressed.We studied all KTRs from 2005 to 2012. 29 (4%), 317 (48%), 217 (33%), 76 (12%), and 21 KTRs (4%) were identified as underweight, normal-weight, overweight, obese, and morbid obese, respectively. 33 of 97 obese KTRs (34%) had pre-existent diabetes. Samples were collected before transplantation and at + 1, + 2, + 3 months posttransplantation. Donor-reactive T-cells were measured using an interferon-γ Elispot assay.Obese KTRs showed an increased incidence pre-existent diabetes (p < 0.001), but no differences for hypertension and coronary artery disease (p > 0.05). Among obese KTRs, those with pre-existent diabetes showed inferior patient and allograft survival, worse allograft function, delayed graft function, and prolonged hospitalization (p < 0.05). Interestingly, no differences were observed between obese non-diabetic, normal-weight diabetic, and normal-weight non-diabetic KTRs (p > 0.05). Obese diabetic KTRs showed higher frequencies of donor-reactive T-cells pretransplantation (p < 0.05).Our results suggest that the increased risk of mortality, allograft loss, delayed graft function, and prolonged hospitalization in obese KTRs is limited to those with diabetes. A state of obesity-related inflammation plus hyperglycemia may trigger increased alloreactivity and should call for adequate immunosuppression.     

Association of genetic polymorphisms of angiopoietin-like 4 with severity of posttransplant proteinuria in kidney allograft recipients

BackgroundProteinuria is a hallmark of glomerular injury, and persistent proteinuria is associated with graft failure in kidney transplant patients. Recently, it is known that the level of circulating angiopoietin-like 4 (ANGPTL4) is elevated in the patients with human nephrotic syndrome, in which ANGPTL4 is responsible for relieving proteinuria.PurposeThe purpose of this study is to determine effects of clinical factors and genetic polymorphism of ANGPTL4 on proteinuria after kidney transplantation.MethodsA total of 282 patients out of 400 renal transplant patients between 2008 and 2012 at St. Vincent Medical Center, CA were studied in a retrospective study design. The level of proteinuria was measured by random urine protein to creatinine ratio, and divided into two groups (Group 1: UPC < 500 mg/day, Group 2: ≥ 500 mg/day). Single nucleotide polymorphisms of ANGPTL4 genes (rs1044250, rs2278236, rs116843064, rs11672433, rs4076317) were determined by real time PCR with sequence specific primers.ResultsAmong various clinical factors, only delayed graft function, mTOR inhibitor use and fish oil use were significantly associated with posttransplant proteinuria. Statistical differences were found in genetic polymorphisms of ANGPTL4 (rs1044250, rs2278236) in regards to proteinuria among tested patients. rs1044250 (C/T, T228M, missense mutation) alleles showed multiple significant differences (Group 1 vs. Group 2: C vs. T: OR = 1.893, CI = 1.322–2.710, p < 0.001). Similar trends were found in rs2278236 (A/G) alleles with statistical significances (Group 1 vs. Group 2: A vs. G: OR = 0.620, CI = 0.443–0.867, p = 0.005). With multiple logistic regression, rs1044250 was still a significant risk factor of moderate/severe proteinuria (p = 0.021).ConclusionsThis study suggests that the presence of C allele of rs1044250 and G allele of rs2278236 in ANGPTL4 gene is associated with higher risk of moderate/severe proteinuria in renal transplant patients.     

Evaluation of tonsillectomy before kidney transplantation in patients with IgA nephropathy

The effectiveness of a tonsillectomy before kidney transplantation (KTx) in suppressing the recurrence of IgA nephropathy (IgAN) has never been studied. The aim of this study was to analyze the effectiveness of a preoperative tonsillectomy for preventing IgAN recurrence and to identify predictive risk factors for IgAN recurrence. Of the 462 recipients who underwent a KTx between 2006 and 2011, a total of 78 patients had biopsy-proven IgAN as their primary disease. Among these 78 patients, 28 patients (group 1) underwent a tonsillectomy and 50 patients (group 2) did not undergo a tonsillectomy before KTx. The time to recurrence was 15.5 ± 8.7 months, in group 1 and 20.2 ± 18.6 months in group 2. No significant difference was observed between the two groups (P = 0.63). Using a multivariate Cox regression analysis, ABO incompatible KTx and acute rejection were associated with a lower incidence of recurrence (P = 0.02 and 0.002 respectively). These results suggested that a preoperative tonsillectomy might not affect the recurrence of IgAN during a short-term follow-up period, whereas preoperative desensitization and the use of a higher steroid dose were effective for suppressing the recurrence of IgAN.     

Giant gallstone performed by emergency laparoscopic cholecystectomy

INTRODUCTIONGallstone disease is very common, but the gallstone bigger than 5 cm in diameter is very rare. It is very challenging to be removed by laparoscopic cholecystectomy (LC) and poses extra difficulty in emergency.PRESENTATION OF CASEA 70-year-old man complained of abdominal pain in the right upper quadrant with fever of 38 °C for two days. Abdominal ultrasound indicated acute cholecystitis and a single, extremely large gallstone (95 mm × 60 mm × 45 mm). Emergency laparoscopic cholecystectomy was performed successfully.DISCUSSIONGallstone over 5 cm in diameter is very rare. LC will be very difficult for these cases, especially for the emergency cases. Emergency laparoscopic cholecystectomy can be successfully performed with clear exposure of the anatomy of the Calot's triangle. To the best of our knowledge, such giant gallstone has been rarely reported.CONCLUSIONWe have proven that for the rare giant gallstone about 10 cm in size, LC is a feasible option if the anatomy of the Calot's triangle can be clearly exposed; otherwise, open cholecystectomy is a safe choice.     

Pretransplant serum BAFF levels are associated with pretransplant HLA immunization and renal allograft survival

BackgroundThe essential function of B cell–activating factor (BAFF) is regulating the survival and differentiation of B cells. The link between pretransplant BAFF levels and pretransplant alloimmunization and its value to predict subsequent acute antibody-mediated rejection (AMR) and outcome after renal transplantation is not fully understood.MethodsObjective of our retrospective single-center study was to determine, by ELISA analysis of pretransplant serum BAFF levels in 249 patients undergoing renal transplantation, association between preformed anti-human leukocyte antigen (HLA) antibodies, occurrence of acute antibody mediated rejection (AMR) and renal allograft survival.ResultsPretransplant serum BAFF levels were significantly higher in presensitized recipients with anti-HLA antibodies (3262 ± 2796 pg/ml) than in recipients without occurrence of anti-HLA antibodies (2252 ± 1425 pg/ml; p < 0.0001). In addition, pretransplant BAFF levels correlated with cumulative MFI values of anti-HLA antibodies (r = 0.2966, p = 0.0025). Patients with high pretransplant BAFF levels (≥ 2137 pg/ml) experienced significantly lower allograft survival rates compared to low pretransplant BAFF levels (80% vs. 91%; p = 0.01). Coexistence of high pretransplant BAFF levels and posttransplant AMR was associated with the worst allograft survival rates (56%). Relative risk (RR) for allograft loss was associated with high serum BAFF levels (RR 2.3; p = 0.02), presence of anti-HLA antibodies (RR 2.5; p = 0.007) or anti-HLA -donor-specific antibodies (DSAs) (RR 2.6; p = 0.003) before transplant and AMR post transplant (RR 2.5; p = 0.007). AMR was the strongest independent risk factor for allograft failure (RR 2.6; p = 0.03).ConclusionElevated pretransplant serum BAFF levels negatively affect renal allograft survival and represent a risk factor for allosensitization and subsequent AMR.     

Is endothelial glycocalyx damage a cause of renal scarring in vesicoureteral reflux with febrile urinary tract infection?

IntroductionEndothelial glycocalyx is a luminal layer which can be damaged by inflammatory agents or pathogens. The endothelial glycocalyx damage is thought to have a role in the formation of renal scars in children who have febrile urinary tract infection and vesicoureteral reflux. This study aimed to compare the blood levels of endothelial glycocalyx components heparan sulfate and Syndecan-1 in children with and without renal scarring due to vesicoureteral reflux-associated febrile urinary tract infection.Materials and methodsData of the patients diagnosed with vesicoureteral reflux without renal scarring (Group 1), patients with vesicoureteral reflux and renal scarring (Group 2), and completely healthy children (Group 3) were retrospectively reviewed. Blood levels of heparan sulfate and Syndecan-1 were measured and the results were compared.ResultsThe entire cohort consisted of 90 patients; there were 30 patients in each group. Mean patient age was 49.7 ± 18.0 months. Mean serum heparan sulfate (42.90 ± 18.90 ng/mL) and Syndecan-1 (37.59 ± 13.77 ng/mL) levels of Group 2 were significantly higher than those of other groups. The cut-off value for heparan sulfate was 35.17 ng/mL, with a 63% sensitivity and 86% specificity. The cut-off value for Syndecan-1 was 29.99 ng/mL with a 70% sensitivity and 80% specificity.ConclusionOur findings indicate that blood levels of heparan sulfate and Syndecan-1 could be related with renal scarring in patients with vesicoureteral reflux, especially in the setting of febrile urinary tract infection. However, due to their low sensitivity, these biomarkers should be used along with clinical data.     

Synergism of a natural plant product,oleanolic acid with calcineurin inhibitor in prolonging islet allograft survival

BackgroundOleanolic acid (OA) is a natural plant-derived triterpenoid with potent anti-inflammatory properties. Since inflammatory cytokines released following islet transplantation hinders engraftment and long-term function, we determined the synergistic ability of OA to with Cyclosporine-A (CSA), a calcineurin inhibitor in improving islet allograft's function and survival.MethodsC57BL/6 mice were rendered diabetic using streptozotocin (200 mg/kg). BALB/c islets were transplanted under the kidney capsule alone (control) or along with administration of OA alone, CSA alone or a combination of both OA and CSA (OA + CSA). T-cell infiltration was analyzed by immunohistochemistry; cytokine concentration was analyzed by Luminex; T-cell cytokine phenotype was analyzed by ELISpot; and alloimmune response was analyzed by flow cytometry.ResultsOA + CSA markedly prolonged islet allograft survival compared to controls (37 ± 5 days vs. 8 ± 3 days). A significant decrease of CD4 + (34 ± 9 vs. 154 ± 42 cells/hpf) and CD8 + T-cellular (46 ± 22 vs. 224 ± 51 cells/hpf, p < 0.0001) infiltration into the graft in OA + CSA treated mice compared to controls. A significant decrease in T cell infiltration was demonstrated in the OA + CSA cohort over either compound application individually. An increase in anti-inflammatory molecules, IL-10 (2.4-fold) and vascular endothelial growth factor (1.6-fold), along with decreased pro-inflammatory cytokines IFN-γ, IL-1β (1.3–2.4-fold) and IL-17 (3.2-fold) was demonstrated. OA + CSA also significantly decreased the frequency of allo-specific T-cell responses. Development of antibodies against donor antigens was also delayed (39 vs 22 days; p < 0.05) in the OA + CSA cohort over administration of either agent individually.ConclusionsOA and CSA exert synergistic effect towards enhancing islet allograft survival and function. This synergistic effect resulted in markedly reduced graft infiltrating cells with attenuation of inflammatory cytokine milieu leading to impairment of both cellular and humoral alloimmune responses. Therefore, novel therapeutic approaches involving combination of OA with calcineurin-inhibitor based immunosuppressant CSA will produce potential beneficial outcomes in clinical islet transplantation.     

Chelerythrine ameliorates acute cardiac allograft rejection in mice

The improvement in graft survival over the past decade has been mainly due to calcineurin inhibitors, which interfere with the calcium-mediated pathway. Recently, other pathways such as those mediated by protein kinase C (PKC) are coming into view. The purpose of this study was to assess the immunosuppressive properties of chelerythrine, a specific PKC inhibitor, in preventing acute rejection in murine heterotopic heart transplantation. Mice were randomly divided into control and chelerythrine treated group. The control group received PBS while the chelerythrine treated group was given intraperitoneal injection doses (1, 5, 10 mg/kg) of chelerythrine from day 0 to day 14 after heart transplantation. Six days after transplantation, cardiac allografts were harvested for further tests. The mean survival time (MST) of the cardiac allograft in untreated animals was 8 days while graft MSTs observed in chelerythrine treated group was 13 and 23 days at 5 and 10 mg/kg treatment doses, respectively (P < 0.05). Histologic assessment of the allograft in chelerythrine group showed a significant decline in histologic rejection score, as well as CD4 + and CD8 + T cell infiltration and ICAM-1 + endothelial cell activation. Down-regulation of Th1/Th2 cytokine expression was observed in chelerythrine treatment group. Meanwhile, chelerythrine was also found to inhibit the dephosphorylation of phosphorylated nuclear factor of activated T cells (NFAT) protein 1 and 4.