ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Orthotopic liver transplantation is the most effective treatment for fulminant hepatic failure. As an alternative treatment, an efficient extracorporeal bioartificial liver should contain a large yield of functional hepatocytes with an immunoprotective barrier, for providing temporary adequate metabolic support to allow spontaneous liver regeneration or for acting as a bridge toward transplantation. Survival, proliferation, and functions of porcine hepatocytes were evaluated in primary cultures and after embedding in alginate beads, which were subsequently coated with a membrane made by a transacylation reaction between propylene glycol alginate and human serum albumin. Disruption of total pig livers by collagenase perfusion/recirculation allowed the obtention of up to 1011hepatocytes with a viability greater than 95%. Hepatocytes in conventional cultures or embedded in coated alginate beads survived for about 10 days, secreted proteins, particularly albumin, and maintained several phase I and II enzymatic activities, namely ethoxyresorufin-O-deethylase, oxidation of nifedipine to pyridine, phenacetin deethylation to paracetamol, glucuroconjugation of paracetamol, and N-acetylation of procainamide. Typical features of mitosis and [3H]thymidine incorporation indicated that porcine hepatocytes proliferated in both conventional cultures and alginate beads. The efficacy of the membrane surrounding alginate beads for protecting cells from immunoglobulins was tested by embedding HLA-typed human lymphocytes, which were subsequently incubated with specific anti-HLA immunoglobulin G and complement. These data show that large yields of porcine hepatocytes that are embedded in coated alginate beads remain functional and are isolated from large molecular weight molecules, such as immunoglobulins. This system represents a promising tool for the design of an extracorporeal bioartificial liver, containing xenogeneic hepatocytes, to treat acute liver disease in humans.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Background.Apoptosis (programmed cell death) occurs as a consequence of global organ ischemia during isolation and storage prior to transplantation. If apoptosis is inhibited during ischemia, organ preservation should be improved, and the length of time for permissible storage may be increased. The objective of this study was to test the effect of a newly developed antiapoptotic compound, LXR-015, during extended hypothermic liver preservation.Methods.Three groups of 12 rats each were studied. In the normal group, liver function was studied immediately after harvesting. In the study group, harvested livers were flushed with Euro-Collins solution (30 ml/kg body weight) containing LXR-015 at a concentration equivalent to 9 mg/kg animal body weight (300 μg/ml). The livers were then stored at 4°C for 24 hr before liver function was studied. In the control group, harvested livers were flushed with Euro-Collins solution without LXR-015 and then stored at 4°C for 24 hr before liver function was studied.Results.Portal venous flow was higher (P<0.05) in the normal and study groups compared with the control group. Portal venous resistance was lower (P<0.05) in the normal and study groups compared with the control group. Liver tissue oxygen consumption in the study group was significantly higher than in both the normal and control groups (P<0.05). Liver enzyme production (aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, creatine kinase) was higher in the control group than in either the study or normal group (P<0.05). Bile production in both the normal and study groups was higher than in the control group (P<0.05). The liver tissue wet to dry weight ratio in both the normal and study groups was lower than in the control group (P<0.05). Histopathology studies revealed fewer apoptotic bodies (P<0.05) in both the normal (1.70±0.15 per high-power field) and study groups (2.08±0.10 per high-power field) than in the control group (7.92±.33 per high-power field).Conclusions.Adding an antiapoptotic compound, LXR-015, to Euro-Collins solution significantly improves hypothermic preservation of the rat liver compared with Euro-Collins solution alone.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Background.The ability to prolong graft survival, in some cases by depleting donor antigen-presenting cells (APCs), and the subsequent demonstration that lymphocytes stimulated by non-APCs become anergic, suggested that graft survival and tolerance induction might be achieved by manipulating donor APCs to render them incompetent. This possibility was tested in histoincompatible murine skin allograft with photodynamic therapy (PDT).Methods.Skin sections (C57BL/6) were exposed in vitro to low doses of benzoporphyrin derivative monoacid ring A (BPD) (verteporfin) and light (λ=690±10 nm; low-dose PDT) before implantation on recipients (BALB/c). Furthermore, the effect of the treatment on the surface molecules of donor-derived Langerhans cells (LC) was evaluated by fluorescence-activated cell sorter analysis; the effect of treatment on the LC alloreactivity in the mixed epidermal cell lymphocyte reaction was also evaluated.Results.Pretreating skin to be grafted with low-dose PDT can significantly prolong the survival of allografts from 9.3±2.2 (n=42) days (control group) to 16.9±1.7 days (n=20; treated group). Moreover, low-dose PDT significantly down-regulated the major histocompatibility complex and costimulatory (B7) molecules (60-90% reduction) on LC, but not LC-specific endocytic receptor (DEC-205), CD45, intercellulr adhesion molecule 1, LC viabilities, and ectophosphatase activity on LC. Additionally, this treatment significantly suppressed the ability of LC to stimulate alloreactive T cells to proliferate.Conclusions.Since engaging T cell receptors in the absence of costimulation results in suboptimal activation of T cells and ultimately anergy, it appears that the immunomodulatory effects of low-dose PDT associated with extended engraftment may depend upon decreased LC expression of major histocompatibility complex and costimulatory molecules.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Human peripheral blood leukocyte (PBL)-reconstituted severe combined immunodeficient (SCID) mice (Hu-PBL-SCID) were used as a model to study xenograft rejection in humans. SCID mice were reconstituted with human PBL using a protocol that included a booster injection with anti-human CD3 antibody-primed cells. This protocol enhanced chimera establishment in SCID mice and resulted in the detection of higher levels of human Ig when compared with SCID mice receiving unprimed PBL alone. Human xenoreactive natural antibodies (XNA), both IgM and IgG subtypes, which recognized porcine islets (PI), were detected in sera of Hu-PBL-SCID by cytofluorometric analysis. Pretreatment of porcine cells with GS-IB4 lectin inhibited the XNA binding, demonstrating the specificity of the XNA from Hu-PBL-SCID. Western blot analysis showed that XNA from normal human serum and Hu-PBL-SCID serum recognized similar xenoantigens on PI, indicating that Hu-PBL-SCID contained a XNA repertoire representative of normal human serum. Immunofluorescent staining of the tissue sections revealed that both human IgG and IgM bound in vivo to the PI engrafted beneath the kidney capsule of Hu-PBL-SCID. In addition, mouse complement (C3) was detected on xenografted PI. The function of xenografted islets were monitored by measuring porcine insulin concentration using a radioimmunoassay. Porcine insulin concentration in the sera of both Hu-PBL-SCID and plain SCID xenografted with PI was similar for up to 14 days after transplantation, after which the insulin levels in Hu-PBL-SCID decreased, thereby indicating rejection. Therefore, PI transplanted into the Hu-PBL-SCID should be a useful model for the study of cellular as well as acquired humoral immune response against xenoislets.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

An intraperitoneally located and prevascularized expanded polytetrafluoroethylene solid support is potentially a suitable transplantation site for encapsulated pancreatic islets, because it allows for both the implantation of a large volume islet graft in the immediate vicinity of blood vessels, and its complete removal. The present study investigates the efficacy of such solid supports for the implantation of nonencapsulated islet isografts in streptozotocin diabetic rat recipients. These solid supports were always coated with acidic fibroblast growth factor, because we found that this growth factor enhances the neovascularization. The success rates of 5-μl (group A) and 10-μl (group B) islet isografts in solid supports were compared with the success rates of 5-μl (group C) and 10-μl (group D) islet isografts implanted in the unmodified peritoneal cavity. Four of seven rats in group A and all seven rats in group B became normoglycemic for at least 6 months. Only two of eight rats in group C and four of eleven rats in group D showed normoglycemia. The normoglycemia lasted for at least 6 months in zero of two animals in group C and in three of four animals in group D. Because of the low success rates in groups C and D, intravenous and oral glucose testing were restricted to the successful recipients in groups A and B. Glucose tolerance was found to be proportional to the grafted islet volume but, expectedly, in both groups the glucose tolerance and the insulin responses were somewhat lower than in controls. Thus, the prevascularized expanded polytetrafluoroethylene solid support, rather than the unmodified peritoneal cavity, is an efficacious transplantation site, potentially suitable for encapsulated islets.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Although simultaneous pancreas and kidney transplantation (SPK) achieves normoglycemia and correction of uremia in type I diabetic patients with renal failure, little data are available on long-term outcome and clinical determinants of recovery of peripheral neuropathy. In this prospective study, 219 electrophysiological studies using a standardized protocol were performed before and up to 8 years after SPK in 44 patients. Nine control diabetic recipients with functioning kidney but nonfunctioning pancreas transplants were studied on 35 occasions. Patients were 38.5±7.9 years old (mean ± SD) with pretransplant diabetes present for 25.2±7.6 years. Significant polyneuropathy (total nerve conduction scores [NCS] <-1.0) was present in 89% before transplantation, which correlated with body weight (r=0.628,P<0.001). Two distinct patterns of neurological recovery were observed after SPK. Conduction velocity (CV) improved in a biphasic pattern, with a rapid initial recovery followed by subsequent stabilization. In contrast, the recovery of nerve amplitude was monophasic, and continued to improve for up to 8 years. Initial improvement in NCS was primarily due to an increase in CV (P=0.002 vs. baseline), and was best in shorter and younger patients. Recovery of total NCS at 6 months after SPK, assessed by multivariate analysis, was least in obese recipients and when performed in patients who had started dialysis before SPK, and was associated with lower transplant kidney isotopic glomerular filtration rate and HLA mismatch (P<0.05 to 0.001). Subsequent improvement was associated with less severe initial neuropathy, smaller body weight, and longer duration of diabetes (P<0.01 to 0.001). Fasting hyperinsulinemia was associated with impairment of initial recovery and subsequent NCS after SPK, but was worse in the control group. Recovery of nerve action potential amplitudes was predicted by better initial amplitudes and HLA mismatch, lower body weight, and the use of nifedipine (P<0.05 to 0.001). Nifedipine was used for hypertension in 33% of SPK and was associated with better CV and amplitudes, particularly in the upper limbs, where there was less neuropathy. The use of angiotensin-converting enzyme inhibitors also appeared beneficial, but this was confined to the lower limbs. SPK resulted in a gradual, sustained, and late improvement in nerve action potential amplitudes, consistent with axonal regeneration and partial reversal of diabetic neuropathy. These data suggest that early transplantation of uremic diabetic patients before onset of severe neuropathy, minimizing obesity and optimizing renal transplant function, maximizes neurological recovery after SPK. Furthermore, the preliminary data support randomized clinical trials for evaluation of nifedipine and angiotensin-converting enzyme inhibitors in diabetic neuropathy.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Background.Vaccination guidelines for transplant recipients include regular boosters of tetanus, diphtheria, and inactivated polio vaccine, but there are few published data on the efficacy of these vaccines in patients receiving immunosuppressive therapy.Methods.Serum antibody values were evaluated before and 4 weeks after tetanus, diphtheria, and inactivated polio vaccination in 164 renal transplant recipients compared with healthy controls. Twelve months later, antibody levels were evaluated in 55 patients.Results.Prebooster tetanus antitoxin values were lower in transplant recipients than in controls. All patients developed protective tetanus antibody levels (≥0.01 IU/ml) after vaccination. After 12 months, serum antibodies had decreased, but all patients maintained protective values. Diphtheria antitoxin titers before and after booster vaccination were lower in patients than in controls: 88.5% of patients and 96.2% of controls developed protective diphtheria antibody values. Twelve months after vaccination, diphtheria antitoxin values were below the protective level (0.1 IU/ml) in 38% of patients. Prebooster antibody values to poliovirus types 1 and 3 were comparable in patients and controls, whereas antibodies to poliovirus type 2 were lower in transplant recipients. Seroprotection rates and geometric mean antibody titers after vaccination were equivalent between the two groups for all three poliovirus types. No difference was observed in antibody levels between patients on different immunosuppressive drug regimens. Adverse reactions were significantly less often reported by transplant recipients.Conclusions.In transplant recipients, tetanus and inactivated polio vaccinations are well tolerated and induce protective antibody levels; diphtheria vaccination as currently recommended is less effective and protective antitoxin values decrease rapidly in these patients within 1 year after vaccination.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Background.Refractory rejection and cyclosporine (CsA)-induced nephropathy remain important causes of renal allograft loss. Previous studies demonstrated that 70-85% of the episodes of refractory acute rejection (AR) occurring in renal allograft recipients on a CsA-based immunosuppressive regimen could be salvaged by conversion to tacrolimus. No data are available regarding the correlation between allograft histology at the time of conversion and the response to tacrolimus. We examined the response to tacrolimus conversion in relation to preconversion biopsies stratified by the Banff criteria.Methods.Since May 1992, we have converted 22 patients from CsA to tacrolimus as part of a rescue protocol. We report on 18 patients in whom 6-month follow-up was available after conversion for biopsyproven AR (n=13) or CsA toxicity (n=5). Sixteen patients were recipients of renal allografts, including three second transplants, and two were recipients of kidney-pancreas transplants. All patients with AR were treated with one or more courses of methylprednisolone and OKT3 before conversion. Renal allograft biopsies were interpreted by a transplant pathologist blinded to the clinical history, and graded according to the Banff criteria. Responses to tacrolimus were scored as improved (creatinine returned to within 150% of baseline), stabilized (creatinine rise arrested), or failed (returned to dialysis).Results.Mean follow-up was 17.3±8 months. Fourteen of 18 patients (78%) showed improvement or stabilization in renal function as assessed by creatinine at 6 months or 1 year (when available). Of the 13 patients with histological AR, nine (69%) improved, including five of six with borderline AR, two of three with grade I AR, and two of four with grade II AR. Of the four other patients with AR, two stabilized and two failed. Three of five patients with severe clinical rejection requiring dialysis (range 2-16 weeks) recovered renal function after conversion. Of five patients with CsA toxicity, two (40%) improved. Seven of eight patients who were converted to tacrolimus less than 90 days after transplantation improved, compared with only 4 of 10 who were converted more than 90 days after transplantation. No grafts were lost in patients with a creatinine ≤3.0 mg/dl at the time of conversion versus two of seven grafts lost when the creatinine was 3.1-5.0 mg/dl and two of eight grafts lost when the creatinine was >5.0 mg/dl.Conclusion.The majority of steroid and antilymphocyte antibody (OKT3 or ATGAM) unresponsive rejections in patients on CsA-based immunosuppression will improve or stabilize after conversion to tacrolimus. There was no correlation with allograft histology stratified by the Banff criteria and the response to tacrolimus. Although there was a trend toward a poorer response with more severe histological rejection, higher serum creatinine at the time of conversion, and longer time from transplantation to conversion, favorable responses were noted in all groups. This indicates that a trial of conversion is warranted, irrespective of the histological severity of injury.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID

摘要:

Background.Hepatitis C virus (HCV) infection causes slowly progressive liver disease. Therefore, the full impact of HCV infection after transplantation may require 5-10 years of follow-up.Methods.We have previously reported the effect of HCV infection, acquired from the donor (study 1) or acquired before transplantation (study 2), on the incidence of liver disease, and on patient and graft survival with a median follow-up of 3-5 years. In study 1, we compared 24 recipients of organs from donors who were positive to antibody to HCV (anti-HCV) to 74 recipients of organs from anti-HCV-negative donors. In study 2, we compared 23 anti-HCV-positive recipients to 80 anti-HCV-negative recipients of kidneys from anti-HCV-negative donors. In this report, we extend the median follow-up period for patient and graft survival to 6-7 years, and discuss the implications of our finding on policies for organ procurement.Results.In study 2, after extended follow-up, there continued to be no significant difference between groups with respect to graft loss, but mortality remained significantly higher among recipients with anti-HCV before transplantation. Compared with recipients without anti-HCV before transplantation, the relative risk of graft loss among recipients with anti-HCV before transplantation was 1.30 (0.66-2.58), the relative risk of death was 2.60 (1.15-5.90), and the relative risk of death due to sepsis was 6.30 (1.99-20). In study 1, after extended follow-up, there continued to be no significant differences between groups, with respect to graft loss or death. Compared with recipients of organs from anti-HCV-negative donors, the relative risk of graft loss among recipients of organs from anti-HCV-positive donors was 0.95 (0.54-1.67), and the relative risk of death was 1.00 (0.49-2.02).Conclusions.The two studies presented in this report provide an apparent paradox, with respect to the impact of HCV infection acquired at the time of transplantation versus before transplantation on posttransplantation clinical outcomes. However, the increased mortality among recipients who acquired HCV infection before transplantation, but not among recipients who acquired HCV at the time of transplantation, could be explained by the longer duration of HCV infection in the former group. These findings are consistent with the known slowly progressive nature of HCV infection. However, in the absence of definitive evidence for an adverse effect on patient or graft survival, we believe that the decision to accept a kidney from an anti-HCV-positive donor should be made by the patient, after discussion with the treating physician.

ISSN:0041-1337    

出版商:OVID    

年代:1997

数据来源: OVID