ABSTRACT
Prior studies of anti-CD40 ligand (CD40L)-based immunosuppression demonstrated effective prevention of islet and kidney allograft rejection in nonhuman primate models; however, clinical development was halted because of thromboembolic complications. An anti-CD40L-specific monoclonal antibody, AT-1501 (Tegoprubart), was engineered to minimize risk of thromboembolic complications by reducing binding to Fcγ receptors expressed on platelets while preserving binding to CD40L. AT-1501 was tested in both a cynomolgus macaque model of intrahepatic islet allotransplantation and a rhesus macaque model of kidney allotransplantation. AT-1501 monotherapy led to long-term graft survival in both islet and kidney transplant models, confirming its immunosuppressive potential. Furthermore, AT-1501-based regimens after islet transplant resulted in higher C-peptide, greater appetite leading to weight gain, and reduced occurrence of cytomegalovirus reactivation compared with conventional immunosuppression. These data support AT-1501 as a safe and effective agent to promote both islet and kidney allograft survival and function in nonhuman primate models, warranting further testing in clinical trials.
Subject(s)
Antibodies, Monoclonal , Kidney , Animals , Ligands , Macaca mulatta , Antibodies, Monoclonal/pharmacology , CD40 Ligand , Macaca fascicularis , AllograftsABSTRACT
When South Florida became a hot spot for COVID-19 disease in March 2020, we faced an urgent need to develop test capability to detect SARS-CoV-2 infection. We assembled a transdisciplinary team of knowledgeable and dedicated physicians, scientists, technologists, and administrators who rapidly built a multiplatform, polymerase chain reaction- and serology-based detection program, established drive-through facilities, and drafted and implemented guidelines that enabled efficient testing of our patients and employees. This process was extremely complex, due to the limited availability of needed reagents, but outreach to our research scientists and multiple diagnostic laboratory companies, and government officials enabled us to implement both Food and Drug Administration authorized and laboratory-developed testing-based testing protocols. We analyzed our workforce needs and created teams of appropriately skilled and certified workers to safely process patient samples and conduct SARS-CoV-2 testing and contact tracing. We initiated smart test ordering, interfaced all testing platforms with our electronic medical record, and went from zero testing capacity to testing hundreds of health care workers and patients daily, within 3 weeks. We believe our experience can inform the efforts of others when faced with a crisis situation.
ABSTRACT
Focal segmental glomerulosclerosis (FSGS) accounts for about 40% of all nephrotic syndrome cases in adults. The presence of several potential circulating factors has been suggested in patients with primary FSGS and particularly in patients with recurrent disease after transplant. Irrespectively of the nature of the circulating factors, this study was aimed at identifying early glomerular/podocyte-specific pathways that are activated by the sera of patients affected by FSGS. Kidney biopsies were obtained from patients undergoing kidney transplantation due to primary FSGS. Donor kidneys were biopsied pre-reperfusion (PreR) and a subset 1-2 hours after reperfusion of the kidney (PostR). Thirty-one post reperfusion (PostR) and 36 PreR biopsy samples were analyzed by microarray and gene enrichment KEGG pathway analysis. Data were compared to those obtained from patients with incident primary FSGS enrolled in other cohorts as well as with another cohort to correct for pathways activated by ischemia reperfusion. Using an ex-vivo cell-based assay in which human podocytes were cultured in the presence of sera from patients with recurrent and non recurrent FSGS, the molecular signature of podocytes exposed to sera from patients with REC was compared to the one established from patients with NON REC. We demonstrate that inflammatory pathways, including the TNF pathway, are primarily activated immediately after exposure to the sera of patients with primary FSGS, while phagocytotic pathways are activated when proteinuria becomes clinically evident. The TNF pathway activation by one or more circulating factors present in the sera of patients with FSGS supports prior experimental findings from our group demonstrating a causative role of local TNF in podocyte injury in FSGS. Correlation analysis with clinical and histological parameters of disease was performed and further supported a possible role for TNF pathway activation in FSGS. Additionally, we identified a unique set of genes that is specifically activated in podocytes when cultured in the presence of serum of patients with REC FSGS. This clinical translational study supports our prior experimental findings describing a potential role of the TNF pathway in the pathogenesis of FSGS. Validation of these findings in larger cohorts may lay the ground for the implementation of integrated system biology approaches to risk stratify patients affected by FSGS and to identify novel pathways relevant to podocyte injury.
Subject(s)
Gene Expression Regulation , Glomerulosclerosis, Focal Segmental/blood , Glomerulosclerosis, Focal Segmental/genetics , Kidney Glomerulus/metabolism , Podocytes/metabolism , Signal Transduction , Adult , Biomarkers/blood , Biopsy , Cohort Studies , Female , Glomerulosclerosis, Focal Segmental/pathology , Humans , Inflammation/genetics , Inflammation/pathology , Kidney Glomerulus/pathology , Male , Podocytes/pathology , Proteinuria/blood , Recurrence , Risk Factors , Signal Transduction/genetics , Treatment OutcomeABSTRACT
PURPOSE: To evaluate the current model of small bowel resection and intestinal transplantation in pigs. METHODS: Forty two Large White pigs were distributed in five groups: G1(n=6), G2(n=6) and G3(n=6) were submitted to 80%,100% and 100% plus right colon resection respectively and G4(n=7) and G5(n=5) to 100% SBR plus IT without and with immunosuppression based on Tacrolimus and Mycophenolic acid. Evaluation included weight control, clinical status, biochemical analysis and endoscopies for graft biopsies. Follow-up in G1 and 2 was 84 days, while in G3, four and five was ± three weeks. RESULTS: G1 increased weight suggesting adaptation while G2 and 3 loused weight and inadequate adaptation. G4 and 5 died of acute cellular rejection (ACR) and sepses respectively. Overall survival in G1, 2, 3, 4 and 5 at 30 days was 100, 100, 0 and 20 %, respectively. Medium survival in G4 and 5 was 14 and 16 days. CONCLUSIONS: The resection of 80% of small intestine in pigs is not suitable for short bowel syndrome induction. Intestinal transplantation with the proposed immunosuppression protocol was effective in prevent the occurrence of severe acute rejection, but inappropriate to increase recipients survival.
Subject(s)
Intestine, Small/transplantation , Models, Animal , Short Bowel Syndrome/surgery , Animals , Biopsy , Body Weight , Cholesterol/blood , Female , Graft Rejection/pathology , Immunosuppression Therapy/methods , Immunosuppressive Agents/blood , Immunosuppressive Agents/therapeutic use , Intestine, Small/pathology , Male , Organ Transplantation/methods , Proteins/analysis , Reproducibility of Results , Short Bowel Syndrome/etiology , Swine , Time Factors , Treatment Outcome , Triglycerides/bloodABSTRACT
PURPOSE: To evaluate the current model of small bowel resection and intestinal transplantation in pigs. METHODS: Forty two Large White pigs were distributed in five groups: G1(n=6), G2(n=6) and G3(n=6) were submitted to 80%,100% and 100% plus right colon resection respectively and G4(n=7) and G5(n=5) to 100% SBR plus IT without and with immunosuppression based on Tacrolimus and Mycophenolic acid. Evaluation included weight control, clinical status, biochemical analysis and endoscopies for graft biopsies. Follow-up in G1 and 2 was 84 days, while in G3, four and five was ± three weeks. RESULTS: G1 increased weight suggesting adaptation while G2 and 3 loused weight and inadequate adaptation. G4 and 5 died of acute cellular rejection (ACR) and sepses respectively. Overall survival in G1, 2, 3, 4 and 5 at 30 days was 100, 100, 0 and 20 %, respectively. Medium survival in G4 and 5 was 14 and 16 days. CONCLUSIONS: The resection of 80% of small intestine in pigs is not suitable for short bowel syndrome induction. Intestinal transplantation with the proposed immunosuppression protocol was effective in prevent the occurrence of severe acute rejection, but inappropriate to increase recipients survival. .
Subject(s)
Animals , Female , Male , Intestine, Small/transplantation , Models, Animal , Short Bowel Syndrome/surgery , Biopsy , Body Weight , Cholesterol/blood , Graft Rejection/pathology , Immunosuppression Therapy/methods , Immunosuppressive Agents/blood , Immunosuppressive Agents/therapeutic use , Intestine, Small/pathology , Organ Transplantation/methods , Proteins/analysis , Reproducibility of Results , Swine , Short Bowel Syndrome/etiology , Time Factors , Treatment Outcome , Triglycerides/bloodABSTRACT
MSC are being explored as a promising novel treatment for SLE. In this study, we: 1) assessed the differential effects of allogeneic versus syngeneic MSC transplantation on lupus-like disease, 2) explored the mechanisms by which MSC modulate disease, and 3) investigated whether lupus-derived-MSC have intrinsic immunomodulatory defects. We showed that in MRL/lpr mice and (NZB/NZW)F1 mice, both B6-MSC and lupus-MSC from young mice ameliorated SLE-like disease and reduced splenic CD3+CD4+ T lymphocytes and CD19+CD21+ B lymphocytes. However, lupus-MSC from older (NZB/NZW)F1 mice did not reduce spleen weights, glomerular IgG deposits, renal pathology, interstitial inflammation, CD3+CD4+ T lymphocytes or CD19+CD21+ B lymphocytes significantly. Thus MSC transplantation ameliorates SLE-like disease partly through decreasing CD4+ T cell and naïve mature B cell numbers. Allogeneic MSC may be preferred over syngeneic lupus-derived-MSC given the decreased overall effectiveness of post-lupus-derived-MSC, which appears partially due to disease and not exclusively intrinsic defects in the MSC themselves.
Subject(s)
B-Lymphocytes/immunology , Kidney Glomerulus/immunology , Lupus Erythematosus, Systemic/therapy , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/immunology , T-Lymphocytes/immunology , Age Factors , Animals , Antigens, CD/biosynthesis , Antigens, CD/immunology , B-Lymphocytes/pathology , Female , Kidney Glomerulus/pathology , Lupus Erythematosus, Systemic/immunology , Lupus Erythematosus, Systemic/pathology , Lymphocyte Count , Mesenchymal Stem Cells/pathology , Mice , Mice, Inbred MRL lpr , Mice, Inbred NZB , Spleen/immunology , Spleen/pathology , T-Lymphocytes/pathology , Transplantation, Homologous , Transplantation, IsogeneicABSTRACT
PURPOSE: To describe the reactive T-cell infiltrate in uveitis and intraocular lymphoma using flow cytometry of clinical intraocular specimens acquired during diagnostic pars plana vitrectomy. METHODS: This was a retrospective review of diagnostic vitreous specimens (1992-2011) obtained at a university-based, tertiary care center. Seventy-eight patients with uveitis or lymphoma undergoing pars plana vitrectomy were selected for intraocular testing based on clinical diagnostic uncertainty. Pars plana vitrectomy with flow cytometry, gene rearrangement studies, and cytology was performed. RESULTS: T-cell infiltrates were found in all diagnostic categories with limited power to discriminate between uveitis and T-lymphocyte reactive infiltrates in response to intraocular lymphoma. Statistically significant differences by two-sample test of means between group means were found between 35 uveitis and 35 B-cell lymphoma cases for T-cell markers CD2, 3, 4, 5, and 7, but not for CD8. The CD4:CD8 ratio had a higher mean value in the uveitis group (P=.0113), and 8 T-cell lymphomas had a statistically greater number of CD3+ lymphocytes compared to uveitis (P=.0199) by two-sample test of means. Likelihood ratios were highest for CD2, CD5, CD7, CD4:CD8 ratio, CD20, and CD22. CONCLUSIONS: Discrimination between uveitis and lymphoma based on cell identification by flow cytometry was limited because of the prevalence of T lymphocytes in all diagnostic categories, emphasizing the importance of a reactive T-cell infiltrate in B-cell lymphomas, which may impede diagnosis. Flow cytometry may allow identification of more cases of T-cell lymphoma than reported when it is combined with gene rearrangement and cytology.
Subject(s)
Flow Cytometry/methods , Intraocular Lymphoma/diagnosis , Lymphoma, B-Cell/diagnosis , Lymphoma, T-Cell/diagnosis , T-Lymphocytes/immunology , Uveitis/diagnosis , Adult , Aged , Aged, 80 and over , Antigens, CD/immunology , Biomarkers/analysis , Female , Gene Rearrangement/genetics , Humans , Immunophenotyping/methods , Intraocular Lymphoma/genetics , Intraocular Lymphoma/immunology , Lymphoma, B-Cell/genetics , Lymphoma, B-Cell/immunology , Lymphoma, T-Cell/genetics , Lymphoma, T-Cell/immunology , Male , Middle Aged , Retrospective Studies , Uveitis/genetics , Uveitis/immunology , Vitreous Body/surgeryABSTRACT
The pathological hallmarks of bronchopulmonary dysplasia (BPD), one of the most common long-term pulmonary complications associated with preterm birth, include arrested alveolarization, abnormal vascular growth, and variable interstitial fibrosis. Severe BPD is often complicated by pulmonary hypertension characterized by excessive pulmonary vascular remodeling and right ventricular hypertrophy that significantly contributes to the mortality and morbidity of these infants. Connective tissue growth factor (CTGF) is a multifunctional protein that coordinates complex biological processes during tissue development and remodeling. We have previously shown that conditional overexpression of CTGF in airway epithelium under the control of the Clara cell secretory protein promoter results in BPD-like architecture in neonatal mice. In this study, we have generated a doxycycline-inducible double transgenic mouse model with overexpression of CTGF in alveolar type II epithelial (AT II) cells under the control of the surfactant protein C promoter. Overexpression of CTGF in neonatal mice caused dramatic macrophage and neutrophil infiltration in alveolar air spaces and perivascular regions. Overexpression of CTGF also significantly decreased alveolarization and vascular development. Furthermore, overexpression of CTGF induced pulmonary vascular remodeling and pulmonary hypertension. Most importantly, we have also demonstrated that these pathological changes are associated with activation of integrin-linked kinase (ILK)/glucose synthesis kinase-3ß (GSK-3ß)/ß-catenin signaling. These data indicate that overexpression of CTGF in AT II cells results in lung pathology similar to those observed in infants with severe BPD and that ILK/GSK-3ß/ß-catenin signaling may play an important role in the pathogenesis of severe BPD.
Subject(s)
Connective Tissue Growth Factor/metabolism , Hypertension, Pulmonary/complications , Lung Injury/complications , Lung Injury/etiology , Pulmonary Alveoli/metabolism , Pulmonary Alveoli/pathology , Animals , Animals, Newborn , Cell Nucleus/metabolism , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Hypertension, Pulmonary/pathology , Lung/blood supply , Lung/enzymology , Lung/pathology , Mice , Models, Biological , Phosphorylation , Pneumonia/complications , Pneumonia/metabolism , Pneumonia/pathology , beta Catenin/metabolismABSTRACT
INTRODUCTION: Proliferative lupus nephritis (LN) is marked by increased renal thromboxane (TX) A2 production. Targeting the TXA2 receptor or TXA2 synthase effectively improves renal function in humans with LN and improves glomerular pathology in murine LN. This study was designed to address the following hypotheses: (1) TXA2 production in the MRL/MpJ-Tnfrsf6(lpr)/J (MRL/lpr) model of proliferative LN is cyclooxygenase (COX)-2 dependent and (2) COX2 inhibitor therapy improves glomerular filtration rate (GFR), proteinuria, markers of innate immune response and glomerular pathology. METHODS: Twenty female MRL/lpr and 20 BALB/cJ mice were divided into 2 equal treatment groups: (1) SC-236, a moderately selective COX2 inhibitor or (2) vehicle. After treatment from the age of 10 to 20 weeks, the effectiveness of inhibition of TXA2 was determined by measuring urine TXB2. Response endpoints measured at the age of 20 weeks were renal function (GFR), proteinuria, urine nitrate + nitrite (NO(x)) and glomerular histopathology. RESULTS: SC-236 therapy reduced surrogate markers of renal TXA2 production during early, active glomerulonephritis. When this pharmacodynamic endpoint was reached, therapy improved GFR. Parallel reductions in markers of the innate immune response (urine NO(x)) during therapy were observed. However, the beneficial effect of SC-236 therapy on GFR was only transient, and renal histopathology was not improved in late disease. CONCLUSIONS: These data demonstrate that renal TXA2 production is COX2 dependent in murine LN and suggest that NO production is directly or indirectly COX2 dependent. However, COX2 inhibitor therapy in this model failed to improve renal pathology, making COX2 inhibition a less attractive approach for treating LN.
Subject(s)
Cyclooxygenase 2 Inhibitors/therapeutic use , Kidney/drug effects , Kidney/metabolism , Lupus Nephritis/drug therapy , Lupus Nephritis/metabolism , Pyrazoles/therapeutic use , Sulfonamides/therapeutic use , Thromboxane A2/biosynthesis , Animals , Female , Glomerular Filtration Rate/drug effects , Kidney/pathology , Lupus Nephritis/pathology , Mice , Mice, Inbred BALB C , Mice, Inbred MRL lpr , Nitrates/urine , Nitric Oxide/biosynthesis , Nitrites/urine , Thromboxane B2/urineABSTRACT
BACKGROUND: Transplantation of more than two livers for recurring graft failure has not been specifically addressed in the literature. METHODS: A retrospective analysis was conducted from a total of 2527 overall liver transplants at our institution. Main indications for multiple retransplant included primary nonfunction, chronic rejection, hepatic artery thrombosis, and recurrent disease. RESULTS: We identified 39 patients who received more than two grafts (32 received 3 grafts, 5 received 4 grafts, and 2 received 5 grafts). All patients required interposition arterial grafts from the aorta and hepatojejunostomy for the biliary reconstruction. Seventeen patients are still alive at last follow-up. Perioperative mortality rates after 3rd, 4th, and 5th liver graft were 25%, 14%, and 50%, respectively. Patient and graft survival rates were 72% and 56% at 1 year, respectively. Median length of stay was 27 days and median graft survival was 2.9 years. CONCLUSIONS: Selection of patients and a significant use of available resources are some of the important factors that clinicians need to take into account when dealing with multiple retransplantations. With such conditions, however, liver retransplantation of more than two grafts can be a life-saving procedure.
Subject(s)
Liver Transplantation/statistics & numerical data , Reoperation/statistics & numerical data , Adult , Child , Gallbladder/surgery , Graft Survival , Hepatitis B/surgery , Hepatitis C/surgery , Humans , Liver Transplantation/methods , Liver Transplantation/mortality , Patient Compliance , Postoperative Complications/epidemiology , Plastic Surgery Procedures/methods , Plastic Surgery Procedures/statistics & numerical data , Recurrence , Reoperation/methods , Reoperation/mortality , Retrospective Studies , Survival Rate , Survivors , Treatment Failure , Treatment OutcomeABSTRACT
Several studies have demonstrated the effectiveness of arginine analog nitric oxide synthase (NOS) inhibitor therapy in preventing and treating murine lupus nephritis. However, MRL/MpJ-FAS (MRL/lpr) mice lacking a functional NOS2 (inducible NOS [iNOS]) gene (NOS2) develop proliferative glomerulonephritis in a fashion similar to their wild-type (wt) littermates. This finding suggests that the effect of arginine analog NOS inhibitors is through a non-iNOS-mediated mechanism. This study was designed to address this hypothesis.NOS2 mice were given either vehicle or a NOS inhibitor (SD-3651) to determine if pharmacological NOS inhibition prevented glomerulonephritis, using wt mice as positive controls. Urine was collected fortnightly to measure albumin. At the time of full disease expression in wt mice, all mice were killed, and renal tissue was examined for light, immunofluorescence, and electron microscopic evidence of disease. Serum was analyzed for anti-double-stranded DNA antibody production.NOS2 mice had higher serum anti-double-stranded DNA antibody antibody levels than those of wt mice. SD-3651 therapy reduced proteinuria, glomerular immunoglobulin G deposition, and electron microscopic evidence of podocytopathy and endothelial cell swelling without affecting proliferative lesions by light microscopy.These studies confirm that genetic iNOS deficiency alone is insufficient to prevent proliferative glomerulonephritis and suggest that iNOS activity may inhibit autoantibody production. These results also suggest that SD-3651 therapy acts via a non-iNOS-mediated mechanism to prevent endothelial cell and podocyte pathology. Studies that elucidate this mechanism could provide a useful drug target for the treatment of nephritis.
Subject(s)
Lysine/analogs & derivatives , Nitric Oxide Synthase Type II/antagonists & inhibitors , Nitric Oxide Synthase Type II/deficiency , Proteinuria/prevention & control , Animals , Antibodies, Antinuclear/blood , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Female , Glomerulonephritis, Membranoproliferative/enzymology , Glomerulonephritis, Membranoproliferative/immunology , Glomerulonephritis, Membranoproliferative/prevention & control , Lupus Nephritis/enzymology , Lupus Nephritis/immunology , Lupus Nephritis/prevention & control , Lysine/pharmacology , Male , Mice , Mice, Inbred C57BL , Mice, Inbred MRL lpr , Mice, Knockout , Nitric Oxide Synthase Type II/genetics , Proteinuria/enzymology , Proteinuria/immunologyABSTRACT
Actions of the lipid mediator thromboxane (Tx) A2 acting through the TP receptor contribute to the pathogenesis of cardiovascular disease. To further explore the role of TxA2 in hypertension, we examined the consequences of deficiency of the TP receptor on the course of hypertension associated with endothelial dysfunction and salt sensitivity. To this end, the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) was administered to TP-deficient (Tp-/-) and wild-type (Tp+/+) control mice in drinking water for 21 wk along with a high-salt (HS; 6% NaCl) diet. Administration of L-NAME increased urinary excretion of TxB2 to a similar extent in both Tp+/+ and Tp-/- animals. L-NAME also caused significant and sustained elevations in blood pressure that reached a maximum between weeks 3 and 6. However, the severity of hypertension was attenuated in the Tp-/- mice throughout the study period (P<0.001). At the end of the study, the wild-type mice developed significant cardiac hypertrophy (23.6+/-2% increase in heart-to-body weight ratio). The severity of cardiac hypertrophy was attenuated in the TP-deficient group (11.1+/-2.6%; P<0.05). In contrast, kidney hypertrophy was exaggerated in the Tp-/- mice compared with controls (37.1+/-5.4 vs. 12.3+/-2.3%; P<0.01). Moreover, the severity of glomerulosclerosis, tubule vacuolization, and interstitial chronic inflammation was also enhanced in the Tp-/- group (P<0.01). Thus, in L-NAME hypertension, TP receptors contribute to elevated blood pressure and cardiac hypertrophy. In this model, TP receptors also provided unexpected protection against kidney injury.
Subject(s)
Enzyme Inhibitors/toxicity , Hypertension, Renal/chemically induced , Hypertension, Renal/metabolism , NG-Nitroarginine Methyl Ester/toxicity , Receptors, Thromboxane A2, Prostaglandin H2/metabolism , Albuminuria/chemically induced , Albuminuria/complications , Albuminuria/metabolism , Animals , Blood Pressure/physiology , Cardiomegaly/complications , Disease Models, Animal , Drinking , Eating , Hypertension, Renal/complications , Isoprostanes/urine , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Nitric Oxide/antagonists & inhibitors , Nitric Oxide/metabolism , Renin-Angiotensin System/physiology , Sodium, Dietary/pharmacology , Thromboxane B2/urineABSTRACT
BACKGROUND: Protocol endoscopy with biopsy is currently the gold standard of small bowel transplantation (SBTx) monitoring, however it is invasive, costly, needs skilled operator, may require anesthesia and may cause complications. We investigated fecal calprotectin level (FCL) as a candidate noninvasive marker for monitoring patients after SBTx. METHODS: A pilot study was performed to test the use of FCL measurement in following up SBTx patients. Ileostomy effluents were collected at various postoperative days before endoscopy and biopsy. FCLs were measured by enzyme-linked immunosorbent assay and a cut-off level of 100 ng/mg was considered positive. The results were retrospectively evaluated in combination with clinical, endoscopic, and histopathological findings. FCLs are presented as median nanogram per milligram. RESULTS: FCLs were measured in 122 samples that were obtained from 29 patients after SBTx. Only 1 of 69 positive FCL did not accompany abnormal findings. Retrospective evaluation showed that 11 samples from six patients (FCL: 217) coincided with rejection episodes, six samples from three patients (FCL: 125) coincided with viral enteritis, 51 samples from 21 patients (FCL: 207) coincided with nonspecific inflammation, 11 samples from two patients (FCL: 998) coincided with chronic intestinal ulceration, and finally 50 samples from 19 patients (FCL: 43) coincided with normal findings. No significant FCL difference was found between rejection, infection, and inflammation. FCL evolution in individuals showed that FCL can predict rejection days before histopathological diagnosis. CONCLUSION: FCL is a sensitive test for ongoing organic intestinal allograft pathologies. It might be useful as prescreening marker to avoid unnecessary endoscopies.
Subject(s)
Feces/chemistry , Intestine, Small/transplantation , Leukocyte L1 Antigen Complex/analysis , Follow-Up Studies , Graft Survival , Humans , Ileostomy , Monitoring, Physiologic/methods , Pilot Projects , Retrospective Studies , Transplantation, Homologous/mortality , Transplantation, Homologous/physiology , Treatment OutcomeABSTRACT
BACKGROUND: The reported incidence of adhesion related small bowel obstruction after abdominal organ transplantation is considerably lower than other abdominal procedures. The purpose of the study was to investigate the influence of immunosuppression on peritoneal adhesion formation after intestinal transplantation in rats. METHODS: Four groups of rats (n = 6) underwent small bowel intestinal transplantation in syngeneic (Groups A, B) and allogeneic (Groups C, D) combinations. Groups B and D received tacrolimus immunosuppression 1 mg/kg/d. Animals were euthanized on postoperative day 7, and the total adhesion score (TAS), tissue hydroxyproline content (HPC), TGF-beta mRNA expression levels and histology were examined. RESULTS: All of the animals in Group C showed severe histological (Grade III) acute cellular rejection. There were no histological signs of rejection in Group D. A significant reduction in TAS was observed in tacrolimus treated animals in both syngeneic and allogeneic combinations (Groups B and D), compared with controls (Groups A and C) (P < 0.001 and P < 0.01, respectively). TAS results correlated with the differences in TGF-beta levels that showed significant reduction when each immunosuppressed group was compared with its nontreated counterpart, i.e., (Groups B versus A, P < 0.05, and Groups D versus C, P < 0.01). TGF-beta levels were significantly high in the rejection group (C) and correlated with the intense adhesion formation that was demonstrated in that group. Group C was also the only group in which a significant elevation in HPC was demonstrated (P < 0.001). CONCLUSION: Intense adhesion formation occurs during early posttransplant acute rejection. Postsurgical adhesion formation is significantly reduced in immunosuppressed rats after intestinal transplantation.
Subject(s)
Immunosuppression Therapy , Intestine, Small/transplantation , Postoperative Complications/prevention & control , Tissue Adhesions/prevention & control , Animals , Hydroxyproline/analysis , Male , RNA, Messenger/analysis , Rats , Rats, Inbred Lew , Transforming Growth Factor beta/genetics , Transplantation, HomologousABSTRACT
We previously described a renal protective effect of factor B deficiency in MRL/lpr mice. Factor B is in the MHC cluster; thus, the deficient mice were H2b, the haplotype on which the knockout was derived, whereas the wild-type littermates were H2k, the H2 of MRL/lpr mice. To determine which protective effects were due to H2 vs factor B deficiency, we derived H2b congenic MRL/lpr mice from the 129/Sv (H2b) strain. Autoantibody profiling using autoantigen microarrays revealed that serum anti-Smith and anti-small nuclear ribonucleoprotein complex autoantibodies, while present in the majority of H2k/k MRL/lpr mice, were absent in the H2b/b MRL/lpr mice. Surprisingly, 70% of MRL/lpr H2b/b mice were found to be serum IgG3 deficient (with few to no IgG3-producing B cells). In addition, H2b/b IgG3-deficient MRL/lpr mice had significantly less proteinuria, decreased glomerular immune complex deposition, and absence of glomerular subepithelial deposits compared with MRL/lpr mice of any H2 type with detectable serum IgG3. Despite these differences, total histopathologic renal scores and survival were similar among the groups. These results indicate that genes encoded within or closely linked to the MHC region regulate autoantigen selection and isotype switching to IgG3 but have minimal effect on end-organ damage or survival in MRL/lpr mice.
Subject(s)
Autoantigens/immunology , Genes, MHC Class I , H-2 Antigens/genetics , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Animals , Autoantibodies/blood , Disease Models, Animal , Glomerulonephritis/genetics , Glomerulonephritis/immunology , Glomerulonephritis/mortality , Immunoglobulin G/blood , Lupus Erythematosus, Systemic/mortality , Mice , Mice, Congenic , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred MRL lprABSTRACT
BACKGROUND: Proliferative lupus nephritis (PLN) is characterized by increased expression of inducible nitric oxide (NO) synthase (iNOS). Inhibition of iNOS with NG-monomethyl L-arginine (L-NMMA) abrogates renal disease in two models of murine PLN, but the mechanism of this effect is unknown. Reactive oxygen species have both direct and indirect pathogenic effects in inflammatory lesions and are therefore potentially an important therapeutic target in PLN. We hypothesized that inhibition of iNOS activity would reduce ROS production in murine PLN. METHODS: A dose escalation of L-NMMA (0, 20, 100, and 500 mg/kg/day) was performed in New Zealand Black x New Zealand White F1 (NZB/W) mice with active renal disease. Twenty-four-hour urine nitrate + nitrite (NOX) was measured with a chemiluminescence NO analyzer. Twenty-four-hour urine 8-isoprostane F2alpha (8-iso-PGF2alpha) was measured by gas chromatography-negative ion chemical ionization mass spectrometry. MRL-MpJFASlpr (MRL/lpr) and NZB/W mice were divided into three groups and given either L-NMMA, L-N6-iminoethyl-lysine (L-NIL), or distilled water for 2 weeks. Urine NOX and 8-iso-PGF2alpha were determined after 2 weeks. RESULTS: L-NMMA reduced both urine NOX and 8-iso-PGF2alpha levels in a dose-dependent fashion in NZB/W and MRL/lpr mice. Urine NOX and 8-iso-PGF2alpha levels were highly correlated. Both specific (L-NIL) and nonspecific (L-NMMA) iNOS inhibition reduced urine NOX and 8-iso-PGF2alpha levels in both models of murine PLN. CONCLUSION: These findings suggest that iNOS activity is a major source of reactive oxidant stress in these models of murine PLN. Future studies will address the pathogenic role of reactive oxygen stress in PLN.
Subject(s)
Enzyme Inhibitors/pharmacology , Lupus Nephritis/drug therapy , Nitric Oxide Synthase Type II/antagonists & inhibitors , Oxidative Stress/drug effects , Animals , Biomarkers/urine , Dinoprost/analogs & derivatives , Dinoprost/urine , Enzyme Inhibitors/administration & dosage , Female , Lupus Nephritis/metabolism , Lysine/analogs & derivatives , Lysine/pharmacology , Mice , Mice, Inbred NZB , Nitrates/urine , Nitrites/urine , omega-N-Methylarginine/administration & dosage , omega-N-Methylarginine/pharmacologyABSTRACT
BACKGROUND: Authors reviewed single center experience of intestinal transplantation for treatment of intra-abdominal neoplastic disease. METHODS: There were 25 auto- or allograft transplantations in 21 patients; desmoid tumor (14), neuroendocrine tumor (2), adenocarcinoma (2), hemangioma (1), lymphoma (1), and solid pseudopapillary tumor (1). Medical records were reviewed for cause of graft loss and mortality, recurrent neoplasm, and quality of life. Survival was analyzed using Kaplan-Meier method. RESULTS: There were 11 graft losses; mortality with functioning graft (6), ischemic necrosis (2), acute or chronic rejection (2), and arterial thrombosis (1) during 38 months of mean follow-up. Seven patients died because of recurrent neoplasm and transplant related complications. Six patients experienced recurrent disease; three desmoid tumor (3/14), two adenocarcinoma (2/2), and one neuroendocrine tumor (1/2). Recurrent desmoid tumors were successfully treated with simple excision. Patient and graft survival in the desmoid tumor are 69.2% and 50.0% at 5 years after transplant. Among 14 survivors, 2 need parenteral nutrition or intravenous hydration. Twelve patients are working full time. CONCLUSIONS: Intestinal transplantation is a reasonable life-saving treatment for catastrophic intra-abdominal neoplastic diseases.
Subject(s)
Abdominal Neoplasms/surgery , Fibromatosis, Abdominal/surgery , Graft Survival , Intestines/transplantation , Adolescent , Adult , Child, Preschool , Duodenum/transplantation , Female , Fibromatosis, Abdominal/pathology , Fibromatosis, Aggressive/pathology , Fibromatosis, Aggressive/surgery , Graft Rejection , Humans , Liver Transplantation , Male , Middle Aged , Pancreas Transplantation , Quality of Life , Recurrence , Stomach/transplantation , Survival Analysis , Transplantation, HomologousABSTRACT
We investigated the role of donor bone marrow cell (DBMC) infusions in immunosuppression withdrawal in adult liver transplantation. Patients enrolled were at least 3 years post-transplantation, with stable graft function. Forty-five (study group: G1) received DBMC, and 59 (control group: G2) did not. Immunosuppression was reduced by one third upon enrollment, by another third the second year of the study and was completely withdrawn the third year. Patient and graft survival were similar between the two groups. Although rejection episodes were significantly less in G1 the first 2 years of the study (35% vs. 57%, p = 0.016), there was no significant difference overall (74% vs. 81%, p = 0.14). Until February 2004, 20 patients, 10 in each group, were immunosuppression free for 1-3 years. Approximately 20% of long-term survivors of liver transplantation can successfully discontinue their immunosuppression. DBMC infusions, do not increase this likelihood.