Transplantation of Cold Stored Porcine Kidneys After Controlled Oxygenated Rewarming

The concept of “controlled oxygenated rewarming” (COR) using ex vivo machine perfusion after cold storage was evaluated as tool to improve renal graft function after transplantation. Renal function after 20 min warm ischemia and 21 h cold storage was studied in an auto‐transplant model in pigs (25–30 kg, n = 6 per group). In the study group, preimplant ex vivo machine perfusion for 90 min was added after cold storage, including gentle warming up of the graft to 20°C (COR). Kidneys that were only cold stored for 21 h served as controls. In vivo follow up was one week; the remaining native kidney was removed during transplantation. COR significantly improved cortical microcirculation upon early reperfusion and reduced free radical mediated injury and cellular apoptosis. Post‐transplant kidney function (peak levels in serum) was also largely and significantly improved in comparison to the control group. A weak inverse correlation was found between renal flow during COR and later peak creatinine after transplantation (r² = 0.5), better values were seen for oxygen consumption, measured during machine perfusion at 20°C (r² = 0.81). Gentle graft rewarming prior to transplantation by COR improves post‐transplant graft outcome and may also be a valuable adjunct in pretransplant graft assessment.     

Bcl-2 protects tubular epithelial cells from ischemia reperfusion injury by inhibiting apoptosis

Ischemia followed by reperfusion leads to severe organ injury and dysfunction. Inflammation is considered to be the most important cause of graft dysfunction in kidney transplantation subjected to ischemia. The mechanism that triggers inflammation and renal injury after ischemia remains to be elucidated; however, cellular stress may induce apoptosis during the first hours and days after transplantation, which might play a crucial role in early graft dysfunction. Bcl-2 is known to inhibit apoptosis induced by the etiological factors promoting ischemia and reperfusion injury. Accordingly, we hypothesized that an augmentation of the antiapoptotic factor Bcl-2 may thus protect tubular epithelial cells by inhibiting apoptosis, thereby ameliorating the subsequent tubulointerstitial injury. We examined the effects of Bcl-2 overexpression on ischemia-reperfusion (I/R) injury using Bcl-2 transgenic mice (Bcl-2 TG) and their wild-type littermates (WT). To investigate the effects of I/R injury, the left renal artery and vein were clamped for 45 min, followed by reperfusion for 0-96 h. Bcl-2 TG exhibited decreased active caspase protein in the tubular cells, which led to a reduction in TUNEL-positive apoptotic cells. Consequently, interstitial fibrosis and phenotypic changes were ameliorated in Bcl-2 TG. In conclusion, Bcl-2 augmentation protected renal tubular epithelial cells from I/R, and subsequent interstitial injury by inhibiting tubular apoptosis.     

Effect of pre-treatment with catecholamines on cold preservation and ischemia/reperfusion-injury in rats

Treatment of organ donors with catecholamines reduces acute rejection episodes and improves long-term graft survival after renal transplantation. The aim of this study was to investigate the effect of catecholamine pre-treatment on ischemia/reperfusion (I/R)- and cold preservation injury in rat kidneys. I/R-injury was induced by clamping the left kidney vessels for 60 min along with a contralateral nephrectomy. Cold preservation injury was induced by storage of the kidneys for 24 h at +4 degrees Celsius in University of Wisconsin solution, followed by syngeneic transplantation. Rats were pre-treated with either dopamine (DA), dobutamine (DB), or norepinephrine (2, 5, and 10 microg/kg/min, each group) intravenously via an osmotic minipump for 24 h before I/R- and cold preservation injury. Pre-treatment with DA (2 or 5 microg/kg/min) and DB (5 microg/kg/min) improved recovery of renal function after I/R-injury and dose dependently reduced mononuclear and major histocompatibility complex class II-positive cells infiltrating the kidney after I/R-injury. One day after I/R-injury, upregulation of transforming growth factor (TGF)-beta 1 and 2 and phosphorylation of p42/p44 mitogen-activated protein kinases was observed in kidneys of animals treated with DA or DB. DA (5 microg/kg/min) and DB (5 microg/kg/min) pre-treatment reduced endothelial cell damage after 24 h of cold preservation. Only DA pre-treatment improved renal function and reduced renal inflammation after 24 h of cold preservation and syngeneic transplantation. Our results demonstrate a protective effect of pre-treatment with catecholamines on renal inflammation and function after I/R- or cold preservation injury. This could help to explain the potent organoprotective effects of catecholamine pre-treatment observed in human kidney transplantation.     

Risk factors for impaired CD4+ T‐cell reconstitution following rabbit antithymocyte globulin treatment in kidney transplantation

To describe long‐term CD4⁺ T‐cell reconstitution after rabbit antithymocyte globulin (rATG) treatment and identify predictive factors following kidney transplantation. A single‐center retrospective study analyzed lymphocyte subsets in rATG‐treated kidney transplant recipients (1986–2009). 589 patients were analyzed (maximum follow‐up 21 years). A comparator group (n = 298) received an anti‐IL‐2 receptor monoclonal antibody. CD4⁺ T‐cell lymphopenia (<200/mm³) was present in 48.5%, 9.2%, 6.7%,2.0%, and 0% of patients at one, three, five, 10, and 20 years post‐transplant, respectively. CD4⁺ T‐cell count increased during the first 10 years but remained below the pretransplant count even after 20 years. At 1, 3, and 6 months post‐transplant, mean CD4⁺ T‐cell count was significantly lower in patients with CD4⁺ T‐cell lymphopenia at 12 months versus patients without lymphopenia. On multivariate analyses, significant independent predictors for long‐term impaired CD4 T‐cell reconstitution were recipient age, pretransplant CD4⁺ T‐cell count, 12‐month CD4⁺ T‐cell count, and tacrolimus or MMF therapy. Recipient age >40 years was identified as a cutoff point. CD4⁺ T‐cell reconstitution following rATG treatment remains impaired even after 21 years. Most risk factors for long‐term impaired CD4⁺ T‐cell reconstitution may be evaluated pretransplant or are modifiable post‐transplant.     

Induction of suppressive allogeneic regulatory T cells via rabbit antithymocyte polyclonal globulin during homeostatic proliferation in rat kidney transplantation

Experimental studies have shown that rabbit antithymocyte polyclonal globulin (ATG) can expand human CD4+CD25++Foxp3+ cells (Tregs). We investigated the major biological effects of a self‐manufactured rabbit polyclonal anti‐rat thymoglobulin (rATG) in vitro, as well as its effects on different peripheral T‐cell subsets. Moreover, we evaluated the allogeneic suppressive capacity of rATG‐induced Tregs in an experimental rat renal transplant model. Our results show that rATG has the capacity to induce apoptosis in T lymphocyte lymphocytes as a primary mechanism of T‐cell depletion. Our in vivo studies demonstrated a rapid but transient cellular depletion of the main T cell subsets, directly proportional to the rATG dose used, but not of the effector memory T cells, which required significantly higher rATG doses. After rATG administration, we observed a significant proliferation of Tregs in the peripheral blood of transplanted rats, leading to an increase in the Treg/T effector ratio. Importantly, rATG‐induced Tregs displayed a strong donor‐specific suppressive capacity when assessed in an antigen‐specific allogeneic co‐culture. All of these results were associated with better renal graft function in rats that received rATG. Our study shows that rATG has the biological capacity immunomodulatory to promote a regulatory alloimmune milieu during post‐transplant homeostatic proliferation.     

冷/热缺血再灌注损伤对大鼠肝脏细胞能量状态、凋亡和胀亡的影响

目的研究冷/热两种条件下肝脏缺血再灌注损伤细胞能量状态与细胞死亡方式的关系,探讨减少此类损伤的措施。方法建立SD大鼠肝门阻断热缺血再灌注模型及大鼠原位肝移植冷缺血再灌注模型,测定术后缺血1h、再灌注1、12、72h肝组织能量物质变化,凋亡、胀亡和坏死细胞,观察组织病理学变化。结果热缺血期肝组织损伤重,肝脏细胞内ATP迅速下降为对照组的21%,肝脏细胞死亡方式以胀亡为主;再灌注后,ATP开始恢复,细胞死亡方式逐渐转为以凋亡为主。而冷缺血肝脏细胞缺血期肝组织损伤相对轻,ATP仅下降为对照组的68%,细胞死亡以凋亡为主;再灌注后12hATP即完全恢复,凋亡也达到顶峰,于72h逐渐下降。相关分析表明ATP、能荷与胀亡细胞数量呈显著负相关(r=-0.774,P<0.01;r=-0.789,P<0.01),与凋亡细胞数量无明显相关。结论(1)缺血再灌注发生后,以能量状态为中心环节,轻度损伤时细胞以凋亡为主,自主清除DNA受损的细胞;损伤严重时则胀亡为主,被动清除受损细胞,提示术前或术中给予能量物质可能有利于减少胀亡及炎症反应,延长肝门阻断时间。(2)单纯以凋亡或胀亡作为衡量组织功能的唯一指标值得探讨。     

Cold preservation mediated renal injury: involvement of mitochondrial oxidative stress

Cold preservation has greatly facilitated the use of cadaveric kidneys for renal transplantation, but, clearly, damage occurs during both the preservation episode and the reperfusion phase (following transplantation). The aims of this study were twofold: to develop an in vivo model that was capable of evaluating renal function at early time points following cold preservation, and to evaluate the extent of renal mitochondrial damage that occurs following short periods of cold preservation in vivo. To accomplish these goals, we developed a novel rat model of in vivo renal cold ischemia followed by warm reperfusion (cold I/R) which avoided the complexity involved with transplantation. Briefly, after a right nephrectomy, cold I/R was initiated via pulsatile perfusion (40 minutes) of the left kidney with a cold University of Wisconsin solution followed by 18 hours of warm reperfusion. Cold I/R resulted in significant renal injury, nitrotyrosine production, and inactivation of the key mitochondrial antioxidant enzyme, manganese superoxide dismutase. Furthermore, the activities of the mitochondrial respiratory complexes were significantly reduced following cold I/R. In conclusion, short-term cold I/R results in inactivation of MnSOD, which may lead to the inhibition of mitochondrial complexes and subsequent renal injury. These data suggest that compounds designed to prevent early mitochondrial injury in kidneys that undergo cold preservation would significantly improve renal function and graft survival following transplantation.     

Glycogen Synthase Kinase-3β Inhibitor Attenuates Renal Damage Through Regulating Antioxidant and Anti-inflammation in Rat Kidney Transplant With Cold Ischemia Reperfusion

BackgroudThe glycogen synthase kinase-3β inhibitor thiadiazolidinone derivative 8 (TDZD-8) has been reported to reduce renal ischemia reperfusion (I/R) injury through inhibiting cell damage. However, it is not known whether TDZD-8 could also play a role in protecting the kidney in rat kidney transplantation with renal cold I/R. The aim of the present study was to explore the possible role of TDZD-8 in protecting renal damage in a cold I/R model of rat kidney transplantation.MethodsThe rat model of kidney transplantation with renal cold I/R was established. The renal tissue pathomorphologic changes, renal function, oxidative stress, and inflammatory response were evaluated by detection of a series of indices by hematoxylin and eosin staining, commercial kits, enzyme-linked immunosorbent assay, and spectrophotofluorometry, respectively.ResultsCompared with I/R and Graft groups, renal function was significantly improved in TDZD and TDZD-G groups, which were accompanied by the reduction of renal injury, oxidative stress, and inflammation.ConclusionsThese results suggest that preconditioning with glycogen synthase kinase-3β inhibitor can attenuate kidney transplantation with renal cold I/R through regulating endogenous antioxidant activity and inflammation.     

Daclizumab Versus Rabbit Antithymocyte Globulin in High‐Risk Renal Transplants: Five‐Year Follow‐up of a Randomized Study

We previously reported a randomized controlled trial in which 227 de novo deceased‐donor kidney transplant recipients were randomized to rabbit antithymocyte (rATG, Thymoglobulin) or daclizumab if they were considered to be at high immunological risk, defined as high panel reactive antibodies (PRA), loss of a first kidney graft through rejection within 2 years of transplantation, or third or fourth transplantation. Patients treated with rATG had lower incidences of biopsy‐proven acute rejection (BPAR) and steroid‐resistant rejection at 1 year. Patients were followed to 5 years posttransplant in an observational study; findings are described here. Treatment with rATG was associated with a lower rate of BPAR at 5 years (14.2% vs. 26.0% with daclizumab; p = 0.035). Only one rATG‐treated patient (0.9%) and one daclizumab‐treated patient (1.0%) developed BPAR after 1 year. Five‐year graft and patient survival rates, and renal function, were similar between the two groups. Overall graft survival at 5 years was significantly higher in patients without BPAR (81.0% vs. 54.8%; p < 0.001). In conclusion, rATG is superior to daclizumab for the prevention of BPAR among high‐immunological‐risk renal transplant recipients. Overall graft survival at 5 years was approximately 70% with either induction therapy, which compares favorably to low‐risk cohorts.     

Donor kidney injury molecule‐1 promotes graft recovery by regulating systemic necroinflammation

Ischemia‐reperfusion injury during kidney transplantation predisposes to delayed graft function, rejection, and premature graft failure. Exacerbation of tissue damage and alloimmune responses may be explained by necroinflammation: an autoamplification loop of cell death and inflammation, which is mediated by the release of damage‐associated molecular patterns (eg, high‐mobility group box‐1; HMGB1) from necrotic cells that activate both innate and adaptive immune pathways. Kidney injury molecule‐1 (KIM‐1) is a phosphatidylserine receptor that is upregulated on injured proximal tubular epithelial cells and enables them to clear apoptotic and necrotic cells. Here we show a pivotal role for clearance of dying cells in regulating necroinflammation in a syngeneic murine kidney transplant model. We found persistent KIM‐1 expression in KIM‐1^+/+ kidney grafts posttransplantation. Compared to recipients of KIM‐1^+/+ kidneys, recipients of KIM‐1^?/? kidneys exhibited significantly more renal dysfunction, apoptosis and necrosis, tubular obstruction, and graft failure. KIM‐1^?/? grafts also had more inflammatory cytokines, infiltrating neutrophils, and macrophages compared to KIM‐1^+/+ grafts. Most significantly, passive release of HMGB1 from apoptotic and necrotic cells led to dramatically higher serum HMGB1 levels and increased proinflammatory macrophages in recipients of KIM‐1^?/? grafts. Our data identify an endogenous protective mechanism against necroinflammation in kidney grafts that may be of therapeutic relevance in transplantation.     

VEGF mitigates histone‐induced pyroptosis in the remote liver injury associated with renal allograft ischemia–reperfusion injury in rats

Clinical evidence has indicated a possible link between renal injury and remote liver injury. We investigated whether extracellular histone mediates remote hepatic damage after renal graft ischemia–reperfusion injury, while vascular endothelial growth factor (VEGF) is protective against remote hepatic injury. In vitro, hepatocyte HepG2 cultures were treated with histone. In vivo, the Brown‐Norway renal graft was stored in 4°C preservation solution for 24 hours and then transplanted into a Lewis rat recipient; blood samples and livers from recipients were harvested 24 hours after surgery. Prolonged cold ischemia in renal grafts enhanced liver injury 24 hours after engraftment. Caspase‐1, ASC, NLRP3, and AIM2 expressions in hepatocyte, CD68⁺‐infiltrating macrophages, tissue, and serum interleukin‐1β and ‐18 were greatly elevated, indicating that pyroptosis occurred in the liver and resulted in acute liver functional impairment. Blocking the caspase‐1 pathway decreased the number of necrotic hepatocytes. VEGF treatment suppressed the hepatocyte pyroptosis and liver function was partially restored. Our data suggested that renal allograft ischemia–reperfusion injury is likely associated with acute liver damage due to hepatocyte pyroptosis induced by histone and such injury may be protected by VEGF administration. VEGF, therefore, may serve as a new strategy against other remote organ injuries related to renal transplantation.     

Attenuation of renal ischemia–reperfusion injury by postconditioning involves adenosine receptor and protein kinase C activation

Significant organ injury occurs after transplantation and reflow (i.e., reperfusion injury). Postconditioning (PoC), consisting of alternating periods of reperfusion and re‐occlusion at onset of reperfusion, attenuates reperfusion injury in organs including heart and brain. We tested whether PoC attenuates renal ischemia–reperfusion (I/R) injury in the kidney by activating adenosine receptors (AR) and protein kinase C (PKC). The single kidney rat I/R model was used. Groups: (1) sham: time‐matched surgical protocol only. In all others, the left renal artery (RA) was occluded for 45 min and reperfused for 24 h. (2) Control: I/R with no intervention at R. All antagonists were administered 5 min before reperfusion. (3) PoC: I/R + four cycles of 45 s of R and 45 s of re‐occlusion before full R. (4) PoC + ARi: PoC plus the AR antagonist 8‐ρ‐(sulfophenyl) theophylline (8‐SPT). (5) PoC + PKCi: PoC plus the PKC antagonist chelerythrine (Che). In shams, plasma blood urea nitrogen (BUN mg/dl) at 24 h averaged 23.2 ± 5.3 and creatinine (Cr mg/dl) averaged 1.28 ± 0.2. PoC reduced BUN (87.2 ± 10 in Control vs. 38.8 ± 9, P = 0.001) and Cr (4.2 ± 0.6 in Control vs. 1.5 ± 0.2, P < 0.001). 8‐SPT and Che reversed renal protection indices after PoC. I/R increased apoptosis, which was reduced by PoC, which was reversed by 8‐SPT and Che. Postconditioning attenuates renal I/R injury by adenosine receptor activation and PKC signaling.     

The loss of renal dendritic cells and activation of host adaptive immunity are long-term effects of ischemia/reperfusion injury following syngeneic kidney transplantation

Ischemia/reperfusion injury associated with kidney transplantation induces profound acute injury, influences early graft function, and affects long-term graft outcomes. To determine whether renal dendritic cells play any role during initial innate ischemia/reperfusion injury and the subsequent development of adaptive immune responses, we studied the behavior and function of renal graft and host infiltrating dendritic cells during early and late phases of renal ischemia/reperfusion injury. Wild type to green fluorescent protein (GFP) transgenic rat kidney transplantation was performed with and without 24-h cold storage. Ischemia/reperfusion injury in cold-stored grafts resulted in histopathological changes of interstitial fibrosis and tubular atrophy by 10 weeks, accompanied by upregulation of mRNAs of mediators of interstitial fibrosis and inflammation. In normal rat kidneys, we identified two populations of renal dendritic cells, predominant CD103(-)CD11b/c(+) and minor CD103(+)CD11b/c(+) cells. After transplantation without cold storage, grafts maintained CD103(-) but not CD103(+) GFP-negative renal dendritic cells for 10 weeks. In contrast, both cell subsets disappeared from cold-stored grafts, which associated with a significant GFP-expressing host CD11b/c(+) cell infiltration that included CD103(+) dendritic cells with a TNF-α-producing phenotype. These changes in graft/host dendritic cell populations were associated with progressive infiltration of host CD4(+) T cells with effector/effector-memory phenotypes and IFN-γ secretion. Thus, renal graft ischemia/reperfusion injury caused graft dendritic cell loss and was associated with progressive host dendritic cell and T-cell recruitment. Renal-resident dendritic cells might function as a protective regulatory network.     

Complement Alternative Pathway Deficiency in Recipients Protects Kidney Allograft From Ischemia/Reperfusion Injury and Alloreactive T Cell Response

Despite the introduction of novel and more targeted immunosuppressive drugs, the long‐term survival of kidney transplants has not improved satisfactorily. Early antigen‐independent intragraft inflammation plays a critical role in the initiation of the alloimmune response and impacts long‐term graft function. Complement activation is a key player both in ischemia/reperfusion injury (IRI) as well as in adaptive antigraft immune response after kidney transplantation. Since the alternative pathway (AP) amplifies complement activation regardless of the initiation pathways and renal IR injured cells undergo uncontrolled complement activation, we speculated whether selective blockade of AP could be a strategy for prolonging kidney graft survival. Here we showed that Balb/c kidneys transplanted in factor b deficient C57 mice underwent reduced IRI and diminished T cell–mediated rejection. In in vitro studies, we found that fb deficiency in T cells and dendritic cells conferred intrinsic impaired alloreactive/allostimulatory functions, respectively, both in direct and indirect pathways of alloantigen presentation. By administering anti‐fB antibody to C57 wt recipients in the early post Balb/c kidney transplant phases, we documented that inhibition of AP during both ischemia/reperfusion and early adaptive immune response is necessary for prolonging graft survival. These findings may have implication for the use of AP inhibitors in clinical kidney transplantation.     

Reduced P-selectin expression on circulating platelets after prolonged cold preservation in renal transplantation

The uremic state in patients with terminal renal insufficiency is accompanied by a bleeding tendency connected with platelet dysfunction. Prolonged cold ischemia and inflammatory interactions between leukocytes, platelets and endothelial cells contribute to ischemia-/reperfusion (I/R) injury and may impair long-term graft survival. We evaluated the influence of the duration of cold preservation time on the expression of platelet GPIIb/IIIa and P-selectin and on the formation of leukocyte-platelet complexes after kidney transplantation. Fourteen patients undergoing kidney transplantation were divided into group I with long preservation time (26.6 +/- 1.9 h) and group II with short preservation time (8 +/- 6.1 h). Five venous blood samples (3 ml) were taken before induction of anesthesia, 12 h, 2, 7 and 14 d after transplantation. Surface expression of the GPIIb/IIIa, P-selectin and the percentage of platelet-granulocyte complexes were quantified by flow cytometry. Additionally blood from seven healthy volunteers was analyzed. GPIIb/IIIa and P-selectin expression on circulating platelets were significantly decreased in the long and the short-term graft preservation group compared with healthy volunteers. A significantly reduced P-selectin expression was found in the long-term preservation group compared with the short-term group. The percentage of platelet-granulocyte complexes also decreased in both preservation groups in the first 2 d after reperfusion and remained in this state in the long-term preservation group. Reduced expression of P-selectin on circulating platelets may be an indicator of I/R injury after prolonged kidney graft preservation.     

Role of combination of L-arginine and α-tocopherol in renal transplantation ischaemia/reperfusion injury: a randomized controlled experimental study in a rat model

What’s known on the subject? and What does the study add? Renal ischaemia/reperfusion (I/R) injury is an inevitable consequence of kidney transplantation. It contributes to delayed graft function (DGF), acute renal failure and graft rejection. The present study investigates for the first time the impact of a combination of l ‐arginine and alpha tocopherol on the renal ischemia/reperfusion injury in a rodent model of kidney transplation. We found that concomitant administration of l ‐arginine and α‐tocopherol has a more protective effect and synergistic antioxidant effect on ischaemia/reperfusion injury in transplanted rat kidneys.

OBJECTIVES

To investigate the role of l ‐arginine and α‐tocopherol in ischaemia/reperfusion injury in a kidney transplanted rat model.

MATERIALS AND METHODS

In total, 40 male Sprague‐Dawley rats subjected to renal transplantation received FK506 (tacrolimus) to overcome early acute rejection episodes. Animals were divided randomly into four groups (ten rats each). Group I were treated with FK506 (2 mg/kg/bw/day) and served as the control group. Group II were treated with l ‐arginine 300 mg/kg/bw. Group III were treated with α‐tocopherol 30 mg/kg/bw. Group IV were treated with l ‐arginine and α‐tocopherol. Urine and blood samples were taken at 0 (before operation), 2, 7 and 14 days post‐transplantation for estimation of urine sodium, creatinine, fractional excretion of sodium, serum creatinine, sodium and blood urea nitrogen. Histological examination and measurement of malondialdehyde in kidney tissues were also performed.

RESULTS

Serum creatinine and blood urea nitrogen significantly decreased in l ‐arginine and α‐tocopherol, as well as combination groups, compared to the control group. Malondialdehyde was significantly decreased in the combination group compared to l ‐arginine and α‐tocopherol alone. Histological examination of the control group showed that acute tubular necrosis was markedly decreased in transplanted kidneys treated with a combination of both l ‐arginine and α‐tocopherol.

CONCLUSIONS

Concomitant administration of l ‐arginine and α‐tocopherol has a more protective effect and synergistic antioxidant effect on ischaemia/reperfusion injury in transplanted rat kidneys.     

Effect of Autologous Adipose-Derived Stem Cells in Renal Cold Ischemia and Reperfusion Injury

Objective

The aim of this study was to investigate the effect of autologous adipose-derived stem cells (ADSCs) on renal cold ischemia and reperfusion (I/R) injury via intravenous infusion on rats.

Methods

A renal cold I/R injury rat model was established. Rats were equally randomized into Sham group, Cold I/R group (cold I/R plus culture medium only), and ADSC-treated group (cold I/R plus immediate intrarenal administration of 2 × 10⁶ autologous ADSCs, followed by intravenous autologous ADSCs 6 hours after reperfusion). All rats were killed 24 hours after the I/R procedure.

Results

Serum creatinine levels were significantly reduced in the ADSC-treated group compared with the Cold I/R group (P < .01). The renal tissue in the ADSC-treated group had well conserved renal architecture compared with the Cold I/R group. The mRNA expression of tumor necrosis factor α was significantly lower and Bcl-2 was higher in the ADSC-treated group than in the Cold I/R group (P < .05).

Conclusions

Autologous ADSC infusions ameliorated renal damage undergoing cold I/R injury and improved the renal function, partly through inhibiting inflammatory reactions and reducing apoptosis.     

Mesenchymal stromal cells and kidney transplantation: pretransplant infusion protects from graft dysfunction while fostering immunoregulation

Bone marrow‐derived mesenchymal stromal cells (MSC) have emerged as useful cell population for immunomodulation therapy in transplantation. Moving this concept towards clinical application, however, should be critically assessed by a tailor‐made step‐wise approach. Here, we report results of the second step of the multistep MSC‐based clinical protocol in kidney transplantation. We examined in two living‐related kidney transplant recipients whether: (i) pre‐transplant (DAY‐1) infusion of autologous MSC protected from the development of acute graft dysfunction previously reported in patients given MSC post‐transplant, (ii) avoiding basiliximab in the induction regimen improved the MSC‐induced Treg expansion previously reported with therapy including this anti‐CD25‐antibody. In patient 3, MSC treatment was uneventful and graft function remained normal during 1 year follow‐up. In patient 4, acute cellular rejection occurred 2 weeks post‐transplant. Both patients had excellent graft function at the last observation. Circulating memory CD8⁺ T cells and donor‐specific CD8⁺ T‐cell cytolytic response were reduced in MSC‐treated patients, not in transplant controls not given MSC. CD4⁺FoxP3⁺Treg expansion was comparable in MSC‐treated patients with or without basiliximab induction. Thus, pre‐transplant MSC no longer negatively affect kidney graft at least to the point of impairing graft function, and maintained MSC‐immunomodulatory properties. Induction therapy without basiliximab does not offer any advantage on CD4⁺FoxP3⁺Treg expansion ( ClinicalTrials.gov number: NCT 00752479).     

Functional analysis of CD4+ CD25bright T cells in kidney transplant patients: improving suppression of donor-directed responses after transplantation

Abstract: Background: The role of CD4⁺ CD25^bright regulatory T cells (Treg) in controlling alloreactivity is established, but little is known whether antigen‐specific Treg are induced in fully immunosuppressed kidney transplant patients. Methods: The frequency and function of CD25^bright T cells of nine stable kidney transplant patients before and 0.5–2 yr after transplantation were measured. Patients received triple therapy consisting of cyclosporine, mycophenolate mofetil and prednisone. To investigate the influence of transplantation and immunosuppression on Treg function, we compared their suppressive capacities pre‐ and post‐transplantation using mixed lymphocyte reactions and kept the CD25^?/dim effector T‐cell (Teff) population constant. Results: After transplantation, the percentage of CD4⁺ CD25^bright T cells significantly decreased from 8.5% pre‐transplant to 6.9% post‐transplant (median, p = 0.05). However, the lower percentage of post‐transplant CD4⁺ CD25^bright T cells was not associated with reduced, but rather improved suppressor function of these cells. The proliferative response of pre‐transplant Teff to donor‐antigens was more profoundly suppressed by post‐transplant Treg than by pre‐transplant Treg (pre‐transplant 18% vs. post‐transplant 55% median, p = 0.03) and was comparable against third party antigens at a CD25^bright:CD25^?/dim ratio of 1:20. Conclusions: In immunosuppressed kidney transplant patients, the donor‐directed suppressive capacity of CD4⁺ CD25^bright regulatory T cells improved, which may contribute to the development of donor‐specific hyporesponsiveness against the graft.