Marlon F Levy

Publications (109)388.56 Total impact

• Article: Tumor biology and pre-transplant locoregional treatments determine outcomes in patients with T3 hepatocellular carcinoma undergoing liver transplantation.

  Peter T W Kim, Nicholas Onaca, Srinath Chinnakotla, Gary L Davis, Linda W Jennings, Greg J McKenna, Richard M Ruiz, Marlon F Levy, Robert Goldstein, Goran B Klintmalm
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  ABSTRACT: Liver transplantation is the optimal treatment for patients with hepatocellular carcinoma (HCC) and cirrhosis. This study was conducted to determine the impact of pre-transplant locoregional therapy (LRT) on HCC and our institution's experience with expansion to United Network of Organ Sharing Region 4 T3 (R4T3) criteria. Two hundred and twenty-five patients with HCC (176 meeting Milan and 49 meeting R4T3 criteria) underwent liver transplantation from 2002 to 2008. Compared with the Milan criteria, HCCs in R4T3 criteria displayed less favorable biological features such as higher median alpha-fetoprotein level (21.9 vs. 8.5 ng/mL, p = 0.01), larger tumor size, larger tumor number, and higher incidence of microvascular invasion (22% vs. 5%, p = 0.002). As a result, patients meeting Milan criteria had better five-yr survival (79% vs. 69%, p = 0.03) and a trend toward lower HCC recurrence rates (5% vs. 13%, p = 0.05). Pre-transplant LRT did not affect post-transplant outcomes in patients meeting Milan criteria but did result in lower three-yr HCC recurrence (7% vs. 75%, p < 0.001) and better three-yr survival (p = 0.02) in patients meeting R4T3 criteria. Tumor biology and pre-transplant LRT are important factors that determine the post-transplant outcomes in patients with HCC who meet R4T3 criteria.
  Clinical Transplantation 01/2013; · 1.67 Impact Factor
• Article: Autoimmune Chronic Pancreatitis with IgG4-Related Pancreatic Pseudocyst in a Patient Undergoing Total Pancreatectomy Followed by Autologous Islet Transplantation: A Case Report.

  Morihito Takita, Takeshi Itoh, Shinichi Matsumoto, Masayuki Shimoda, Daisuke Chujo, Shuichi Iwahashi, Yoshiko Tamura, Nicholas Onaca, Bashoo Naziruddin, Bryan L Bartlett, Marlon F Levy
  Pancreas 01/2013; 42(1):175-177. · 2.39 Impact Factor
• Article: Sirolimus and Cardiovascular Disease Risk in Liver Transplantation.

  Greg J McKenna, James F Trotter, Erik Klintmalm, Richard Ruiz, Nicholas Onaca, Giuliano Testa, Giovanna Saracino, Marlon F Levy, Robert M Goldstein, Goran B Klintmalm
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  ABSTRACT: BACKGROUND: Two adverse effects of sirolimus are hypertriglyceridemia and hypercholesterolemia. These elevated levels often lead clinicians to discontinue the sirolimus from concerns of an increased cardiovascular disease (CVD) risk; however, evidence suggests that sirolimus might be cardioprotective. There are no published reports of sirolimus CVD in liver transplantation. METHODS: We reviewed all 1812 liver recipients who underwent transplantation from 1998 to 2010, identifying a cohort using sirolimus as part of the initial immunosuppression (SRL Cohort) and a control group of the remaining patients from this period where SRL was never given (Non-SRL Control). A prospectively maintained database identified all episodes of myocardial infarction (MI), congestive heart failure (CHF), abdominal aortic aneurysm (AAA), and cerebrovascular accident and tracked triglyceride, high-density and low-density lipoproteins, and total cholesterol levels. A Framingham Risk Model calculated the predicted 10-year risk of CVD for both groups. RESULTS: The SRL Cohort (n=406) is older, more predominantly male, with more pretransplantation hypertension and diabetes and posttransplantation hypertension compared to Non-SRL Controls (n=1005). The SRL Cohort has significantly higher triglyceride, low-density lipoprotein, and cholesterol levels at 6 months and 1 year. There is no difference in MI incidence in the SRL Cohort (1.0% vs. 1.2%) and no difference in AAA, cerebrovascular accident, and CHF. The Framingham Risk Model predicts that the SRL Cohort should have almost double the 10-year risk of CVD compared to the Non-SRL Control (11% vs. 6%). CONCLUSIONS: Sirolimus causes hypertriglyceridemia and hypercholesterolemia, but it does not increase the incidence of MI or other CVDs. Considering the SRL Cohort has more cardiac risk factors and nearly double 10-year predicted CVD risk, the fact that the CVD incidence is similar suggests that sirolimus is in fact cardioprotective.
  Transplantation 12/2012; · 4.00 Impact Factor
• Article: Elevation of High-Mobility Group Box 1 after Clinical Autologous Islet Transplantation and Its Inverse Correlation with Outcomes.

  Takeshi Itoh, Shuichi Iwahashi, Mazhar A Kanak, Masayuki Shimoda, Morihito Takita, Daisuke Chujo, Yoshiko Tamura, Ana M Rahman, Wen Y Chung, Nicholas Onaca, P Toby H Coates, Ashley R Dennison, Bashoo Naziruddin, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: A major problem after clinical autologous islet transplantation (AIT) is the difficulty in achieving insulin independence. To follow up on our demonstration in a murine model that high-mobility group box 1 (HMGB1) was released from islets and involved in early loss of transplanted islets, we tested the role of HMGB1 in clinical AIT. Serum HMGB1 levels from 15 AIT patients were significantly elevated during islet infusion (7.6 ± 1.2 ng/mL) and 24 hours after infusion (8.0 ± 1.4 ng/mL) compared to admission levels (2.4 ± 0.6 ng/mL). The first elevation of HMGB1 was associated with islet damage, but the later elevation was not. The change in the HMGB1 level from admission to first peak (ΔHMGB1) was significantly higher in the AIT group (8.1 ± 1.1 ng/mL) than in the pancreatectomy-only control (2.2 ± 0.5 ng/mL) (p<0.05). Circulating serum levels of soluble receptor for advanced glycation end products (sRAGE) were also elevated during islet infusion. In vitro studies demonstrated damaged human islets released HMGB1 but not sRAGE. In terms of outcomes, the insulin free group showed significantly lower ΔHMGB1 (5.2 ± 0.6 ng/mL) and higher ΔsRAGE (2.3 ± 0.6 ng/mL) than the insulin-dependent group (10.6 ± 1.9 ng/mL and 0.7 ± 0.2 ng/mL. respectively). The ΔHMGB1 correlated with the number of white blood cell, IP-10, EGF and Eotaxin. In conclusion, serum HMGB1 was elevated in AIT, and could be associated with inflammatory reactions that deteriorate islet engraftment. Therefore, anti-HMGB1 therapy might be a candidate for further improving the outcomes of clinical AIT.
  Cell Transplantation 12/2012; · 5.13 Impact Factor
• Article: An effective purification method using large bottles for human pancreatic islet isolation.

  Masayuki Shimoda, Takeshi Itoh, Shuichi Iwahashi, Morihito Takita, Koji Sugimoto, Mazhar A Kanak, Daisuke Chujo, Bashoo Naziruddin, Marlon F Levy, Paul A Grayburn, Shinichi Matsumoto
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  ABSTRACT: The purification process is one of the most difficult procedures in pancreatic islet isolation. It was demonstrated that the standard purification method using a COBE 2991 cell processor with Ficoll density gradient solution harmed islets mechanically by high shear force. We reported that purification using large bottles with a lower viscosity gradient solution could improve the efficacy of porcine islet purification. In this study, we examined whether the new bottle purification method could improve the purification of human islets. Nine human pancreata from brain-dead donors were used. After pancreas digestion, the digested tissue was divided into three groups. Each group was purified by continuous density gradient using ET-Kyoto and iodixanol gradient solution with either the standard COBE method (COBE group) or the top loading (top group) or bottom loading (bottom group) bottle purification methods. Islet yield, purity, recovery rate after purification, and in vitro and in vivo viability were compared. Islet yield per pancreas weight (IE/g) and the recovery rate in the top group were significantly higher than in the COBE and bottom groups. Furthermore, the average size of purified islets in the top group was significantly larger than in the COBE group, which indicated that the bottle method could reduce the shear force to the islets. In vivo viability was also significantly higher in the top group compared with the COBE group. In conclusion, the top-loading bottle method could improve the quality and quantity of human islets after purification.
  Islets 11/2012; 4(6).
• Article: Emergence of a Broad Repertoire of GAD65-specific T Cells in Type 1 Diabetes Patients with Graft Dysfunction after Allogeneic Islet Transplantation.

  Daisuke Chujo, Emile Foucat, Morihito Takita, Takeshi Itoh, Koji Sugimoto, Masayuki Shimoda, Kunimasa Yagi, Masakazu Yamagishi, Yoshiko Tamura, Liping Yu, Bashoo Naziruddin, Marlon F Levy, Hideki Ueno, Shinichi Matsumoto
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  ABSTRACT: Islet transplantation is one of the most promising therapies for Type 1 diabetes (T1D). A major issue in islet transplantation is the loss of graft function at late phase. Several studies suggested the involvement of islet-specific T cells in such islet graft dysfunction. In this study, we investigated the breadth and type of glutamic acid decarboxylase 65 (GAD65)-specific T cells in T1D patients after allogeneic islet transplantation. Peripheral blood mononuclear cells (PBMCs) were obtained from islet-transplanted T1D patients during insulin independent period, and cultured for 7 days with pools of GAD65 overlapping peptides in the presence of IL-2. Cytokine secretion profiles of peptide-reactive T cells were analyzed after a short-term re-stimulation with the same peptides, by a multiplex bead-based cytokine assay and by an intracytoplasmic cytokine detection assay. Robust GAD65-specific CD4⁺ and CD8⁺ T cell responses were detected in patients who eventually developed chronic graft dysfunction. Multiple GAD65 peptides were found to induce specific T cell responses in these patients, indicating that the repertoire of GAD65-specific T cells was broad. Furthermore, GAD65-specific CD4⁺ T cells were composed of heterogeneous populations which differentially expressed cytokines including IFN-γ and Type 2 cytokines, but not IL-10. In contrast, patients who showed only marginal GAD65-specific T cell responses maintained substantially longer graft survival and insulin independence. In conclusion, our study suggests that the emergence of islet-specific T cells precedes the development of chronic graft dysfunction in islet-transplanted patients. Thus, our observations support the hypothesis that these islet-specific T cells contribute to the development of chronic islet graft dysfunction.
  Cell Transplantation 09/2012; · 5.13 Impact Factor
• Article: Safety and tolerability of the T-cell depletion protocol coupled with anakinra and etanercept for clinical islet cell transplantation.

  Morihito Takita, Shinichi Matsumoto, Masayuki Shimoda, Daisuke Chujo, Takeshi Itoh, Jeffrey A Sorelle, Kerri Purcell, Nicholas Onaca, Bashoo Naziruddin, Marlon F Levy
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  ABSTRACT: Islet cell transplantation (ICT) is a promising approach to cure patients with type 1 diabetes. We have implemented a new immunosuppression protocol with antithymoglobulin plus anti-inflammatory agents of anakinra and eternacept for induction and tacrolimus plus mycophenolate mofetil for maintenance [T-cell depletion with anti-inflammatory (TCD-AI) protocol], resulting in successful single-donor ICT. Eight islet recipients with type 1 diabetes reported adverse events (AEs) monthly. AEs were compared between three groups: first infusion with the TCD-AI protocol (TCD-AI-1st) and first and second infusion with the Edmonton-type protocol (Edmonton-1st and Edmonton-2nd). The incidence of symptomatic AEs within the initial three months in the TCD-AI-1st group was less than in the Edmonton-1st and Edmonton-2nd groups, with a marginally significant difference (mean ± SE: 5.5 ± 0.3, 7.5 ± 0.5, and 8.3 ± 1.3, respectively; p = 0.07). A significant reduction in liver enzyme elevation after ICT was found in the TCD-AI-1st group compared with the Edmonton-1st and Edmonton-2nd groups (p < 0.05). Because of AEs, all patients in the Edmonton protocol eventually converted to the TCD-AI protocol, whereas all patients tolerated the TCD-AI protocol. TCD-AI protocol can be tolerated for successful ICT, although this study includes small cohort, and large population trial should be taken.
  Clinical Transplantation 09/2012; 26(5):E471-84. · 1.67 Impact Factor
• Article: Gene expression changes in human islets exposed to type 1 diabetic serum.

  Andrew M Jackson, Mazhar A Kanak, Ellen Grishman, Damien Chaussabel, Marlon F Levy, Bashoo Naziruddin
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  ABSTRACT: A major obstacle to the success of islet cell transplantation as a standard treatment for labile type 1 diabetes mellitus is the immediate loss of up to 70% of the transplanted islet mass. Activation of the complement cascade and coagulation factors has been implicated in initiating the destruction of the islet graft. In this study, we analyzed the gene expression changes in islet cells following exposure to type 1 diabetes mellitus serum (T1DM). Isolated human pancreatic islet cells were cultured for 2 d to stabilize islet cell gene expression. Cultured islets were divided into three groups for treatment as follows: group 1 was treated with autologous donor serum, while groups two and three were treated with sera from ABO-matched allogeneic donors or autoantibody positive type 1 diabetic patient, respectively. Complement was detected using anti-C3 FITC and CH50 assay. Islet gene expression was analyzed using Illumina micro-array technology. Results were confirmed using real-time PCR. Immunofluorescent imaging demonstrated complement deposition only in the T1DM condition. Gene array and class prediction analysis generated a list of 50 genes that were able to predict the effect of T1DM serum on islets. Quantitative PCR corroborated microarray results. Both techniques demonstrated upregulation of MMP9 (243%), IL-1β (255%), IL-11 (220%), IL-12A (132%), RAD (343%) and a concomitant downregulation of IL-1RN (64%) in islets treated with T1DM serum. Islets treated with T1DM serum overexpressed genes associated with angiogenesis while decreasing transcription of genes that protect islets from inflammatory cytokines and reactive oxygen species.
  Islets 07/2012; 4(4).
• Article: Correlation of Released HMGB1 Levels with the Degree of Islet Damage in Mice and Humans and with the Outcomes of Islet Transplantation in Mice.

  Takeshi Itoh, Morihito Takita, Jeffrey A Sorelle, Masayuki Shimoda, Koji Sugimoto, Daisuke Chujo, Huanying Qin, Bashoo Naziruddin, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: Establishing reliable islet potency assay is a critical and unmet issue for clinical islet transplantation. Recently, we reported that islets contained high levels of high-mobility group box 1 (HMGB1) and damaged islets released HMGB1 in a mouse model. In this study, we hypothesized that the amount of released HMGB1 could reflect the degree of islet damage, and could predict the outcome of islet transplantation. Four groups of damaged mouse islets and three groups of damaged human islets were generated by hypoxic conditions. These islets were assessed by in vivo (transplantation) and in vitro (released HMGB1 levels, released Cpeptide levels, PI staining, TUNEL staining, ATP/DNA and glucose-stimulated insulin release test) assays. In addition, the ability of each assay to distinguish between non-cured (n=13) and cured (n=7) mice was assessed. The curative rates of STZ-diabetic mice after receiving control, hypoxia-3hr, hypoxia-6hr and hypoxia-24hr mouse islets were 100%, 40%, 0%, and 0%, respectively. Only amounts of released HMGB1 and ratio of PI staining significant increased according to the degree of damages in both human and mouse islets. In terms of predictability of curing diabetic mice, amounts of released HMGB1 showed the best sensitivity (100%), specificity (100%), positive (100%) and negative predictive values (100%) among all the assays. The amount of released HMGB1 reflected the degree of islet damage and correlated with the outcome of islet transplantation in mice. Hence, released HMGB1 levels from islets should be a useful marker to evaluate the potency of isolated islets.
  Cell Transplantation 04/2012; · 5.13 Impact Factor
• Article: Low Temperature Condition Prevents Hypoxia Induced Islet Cell Damage and HMGB1 Release in a mouse model.

  Takeshi Itoh, Koji Sugimoto, Morihito Takita, Masayuki Shimoda, Daisuke Chujo, Jeff A Sorelle, Bashoo Naziruddin, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: One of the major issues in clinical islet transplantation is the poor efficacy of islet isolation. During pancreas preservation and islet isolation, islets suffer from hypoxia as islets are highly sensitive to hypoxic conditions. Cold preservation has been applied to minimize hypoxia induced cell damage during organ preservation. However, the studies related to hypoxia induced islet cell damage during islet isolation are limited. Recently, we demonstrated that mouse islets contain high levels of high-mobility group box 1 protein (HMGB1) and during pro-inflammatory cytokine-induced damage, islets release HMGB1 outside the cell. The released HMGB1 is involved in the initial events of early islet loss. In the present study, we hypothesize that low temperature conditions could prevent both hypoxia induced islet cell damage and HMGB1 release from islets in a mouse model. Isolated mouse islets underwent normoxic condition (95% air and 5% CO2) at 37(o)C or hypoxic conditions (1% O2, 5% CO2 and 94% N2) at 37(o)C (hypoxia-37(o)C islets), 22 (o)C (hypoxia-22(o)C islets) or 4(o)C (hypoxia-4(o)C islets) for 12 hours. In vitro and in vivo viability and functionality tests were performed. HMGB1, IL-6, G-CSF, KC, RANTES, MCP-1 and MIP-1α levels in the medium were measured. Low temperature conditions substantially reduced hypoxia induced necrosis (p<0.05) and apoptosis (p<0.05). In addition, low temperature islet culture significantly increased the insulin secretion from islets by high glucose stimulation (p<0.05). All of recipient mice reversed diabetes after receiving the hypoxia-4(o)C islets but not after receipt of hypoxia-37(o)C or 22(o)C islets. The amount of released HMGB1, IL-6, G-CSF, KC, RANTES, MCP-1 and MIP-1α were significantly reduced in the hypoxia-4(o)C islets compared to those of the hypoxia-37(o)C islets (p<0.05). In conclusion, low temperature conditions could prevent hypoxia-induced islet cell dmage, inflammatory reactions in islet, HMGB1 release and expression. Low temperature conditions should improve the efficacy of isolated islets.
  Cell Transplantation 04/2012; · 5.13 Impact Factor
• Article: Improvement of collagenase distribution with the ductal preservation for human islet isolation.

  Masayuki Shimoda, Takeshi Itoh, Koji Sugimoto, Shuichi Iwahashi, Morihito Takita, Daisuke Chujo, Jeffery A Sorelle, Bashoo Naziruddin, Marlon F Levy, Paul A Grayburn, Shinichi Matsumoto
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  ABSTRACT: A delivery of collagenase at the islet-exocrine interface is crucial for successful human islet isolation. In this study, we investigated how the ductal preservation method at the procurement site affected collagenase distribution. At first, we analyzed human islet isolation data among groups using Serva collagenase with or without ductal injection (DI) or using new Liberase MTF with DI. Then, to assess the distribution of collagenase, human pancreata were classified into two groups: without DI (no DI, n = 5) and with DI at the procurement site (DI, n = 5). Collagenase with 1% marking dye was perfused in the same manner as in our clinical isolation. The distension of the pancreas and the microscopic distribution of the dyed collagenase in pancreas sections were examined. For microscopic analysis, islets were counted and classified into three criteria: unreached, dye didn't reach the islet surface; surface, dye resided on the surface of the islet but not inside; and inside, dye was found inside the islet. As a result, DI groups substantially improved islet yields. In addition, Liberase MTF with DI significantly improved efficacy of pancreas digestion. All pancreata were well distended macroscopically. However, microscopically, the majority of islets in the no DI group were untouched by the dyed collagenase. Ductal preservation substantially improved dyed collagenase delivery on the surface of islets. In conclusion, delivery of collagenase on the surface of islets was unexpectedly insufficient without DI, which was substantially improved by DI. Thus, ductal preservation is a potent method to improve collagenase delivery and islet yields.
  Islets 03/2012; 4(2):130-7.
• Article: Impact of Tissue Volume and Purification on Clinical Autologous Islet Transplantation for the Treatment of Chronic Pancreatitis.

  Shinichi Matsumoto, Morihito Takita, Masayuki Shimoda, Koji Sugimoto, Takeshi Itoh, Daisuke Chujo, Jeffery A Sorelle, Yoshiko Tamura, Rahman M Ana, Nicholas Onaca, Bashoo Naziruddin, Marlon F Levy
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  ABSTRACT: Autologous islet transplantation after total pancreatectomy is an excellent treatment for painful chronic pancreatitis. Traditionally, islets have been isolated without purification; however, purification is applied when the tissue volume is large. Nevertheless, the impact of tissue volume and islet purification on clinical outcomes of autologous islet transplantation has not been well examined. We analyzed 27 cases of autologous islet transplantation performed from October 2006 to January 2011. After examining the relationship between tissue volume and portal pressure at various time points, we compared islet characteristics and clinical outcomes between cases with complications (complication group) and without (noncomplication group), as well as cases with purification (purification group) and without (nonpurification group). Tissue volume significantly correlated with maximum (R = 0.61), final (R = 0.53), and delta (i.e., difference between base and maximum; R = 0.71) portal pressure. The complication group had a significantly higher body mass index, tissue volume, islet yield, and portal pressure (maximum, final, delta), suggesting that complications were associated with high tissue volume and high portal pressure. Only one of four patients (25%) in the complication group became insulin free, whereas 11 of 23 patients (49%) in the noncomplication group became insulin free with smaller islet yields. The purification group had a higher islet yield and insulin independence rate but had similar final tissue volume, portal pressure, and complication rates compared with the nonpurification group. In conclusion, high tissue volume was associated with high portal pressure and complications in autologous islet transplantation. Islet purification effectively reduced tissue volume and had no negative impact on islet characteristics. Therefore, islet purification can reduce the risk of complications and may improve clinical outcome for autologous islet transplantation when tissue volume is large.
  Cell Transplantation 02/2012; · 5.13 Impact Factor
• Article: Islet purification method using large bottles effectively achieves high islet yield from pig pancreas.

  Masayuki Shimoda, Hirofumi Noguchi, Yasutaka Fujita, Morihito Takita, Tetsuya Ikemoto, Daisuke Chujo, Bashoo Naziruddin, Marlon F Levy, Naoya Kobayashi, Paul A Grayburn, Shinichi Matsumoto
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  ABSTRACT: Porcine islets are a promising resource for xenotransplantation. However, low efficacy of islet isolation because of their marked fragility remains a problem. Recently we found that the standard purification method using COBE 2991 cell processor (COBE) with Ficoll density gradient solution damaged islets mechanically by high shearing force. In this study, we evaluated our new purification method using large plastic bottles for the efficacy of islet purification. Ten porcine pancreata were used. The average warm ischemic time was over 40 min; therefore, these pancreata were considered to be in a marginal condition. After digestion, the digested tissue was divided into three groups. Each group was purified using either top loading method with bottle (top group) or bottom loading method with bottle (bottom group) or standard COBE method (COBE group). Islet yield per pancreas weight (IEQ/g) and the rate of postpurification recovery in the top group were significantly higher than the COBE group (top: 8060 ± 1652 IEQ/g, bottom: 4572 ± 614 IE/g, COBE: 3900 ± 734 IE/g. p < 0.02 in top vs. COBE; top percentage of recovery: 99.3 ± 12.3%, bottom: 62.6 ± 8.8%, COBE: 49.5 ± 6.7%, p < 0.02 in top vs. bottom and COBE). The average sizes of purified islets in the top and bottom groups were significantly larger than COBE group (Average diameter top: 156 ± 8 μm, bottom: 147 ± 6 μm, COBE: 119 ± 6 μm, p < 0.01 in top vs. COBE and in bottom vs. COBE), which indicated that bottle method can reduce shear force during purification. Our new purification using top loading bottle method enabled us to obtain a high yield of porcine islets from marginal pancreata.
  Cell Transplantation 01/2012; 21(2-3):501-8. · 5.13 Impact Factor
• Article: Adverse events in clinical islet transplantation: one institutional experience.

  Morihito Takita, Shinichi Matsumoto, Hirofumi Noguchi, Masayuki Shimoda, Tetsuya Ikemoto, Daisuke Chujo, Yoshiko Tamura, Greg S Olsen, Bashoo Naziruddin, Kerri Purcell, Nicholas Onaca, Marlon F Levy
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  ABSTRACT: Islet transplantation is one of the most promising treatments for an unstable form of type 1 diabetes. However, islet transplantation still has some obstacles, such as low success rate of islet isolation, difficulty to obtain long-term insulin freedom, and adverse events related to transplant protocol. We describe the adverse events of current clinical islet transplantation at our institute in this report. Nine type 1 diabetic patients received 17 islet infusions from March 2005 to October 2008. The islet infusion procedure and immunosuppression regimen were based on a modified Edmonton protocol. Severe adverse events (SAEs) were defined as events that were more than grade 3 according to the Terminology Criteria for Adverse Events in Trials of Adult Pancreatic Islet Transplantation, version 4.1 (Collaborative Islet Transplant Registry, CITR). Sixteen events were reported as SAEs and among them 12 events were probably or definitely related to transplant protocols; all occurred within 1 year after infusion except for one. Five adverse events (31%) occurred within 10 days after transplantation and were related to infusion procedures. Seven events (44%) occurred after 50 days and were related to immunosuppressive therapy. SAEs related to the protocol included three events of elevated liver enzymes, two of hemorrhage into gall bladder or peritoneal cavity, two of neutropenia, two of infection, one of vomiting, one of diarrhea, and one of renal dysfunction. All events were grade 3, except for one case that was grade 4 of neutropenia. All SAEs resolved with no sequelae. Neoplasms and deaths were not observed in our study. The present study suggests need to improve both infusion procedure and immunosuppressive strategy from the view of preventing SAEs.
  Cell Transplantation 01/2012; 21(2-3):547-51. · 5.13 Impact Factor
• Article: Improved pancreatic islet isolation outcome in autologous transplantation for chronic pancreatitis.

  Bashoo Naziruddin, Shinichi Matsumoto, Hirofumi Noguchi, Morihito Takita, Masayuki Shimoda, Yasutaka Fujita, Daisuke Chujo, Chad Tate, Nicholas Onaca, Jeffrey Lamont, Naoya Kobayashi, Marlon F Levy
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  ABSTRACT: Total or partial pancreatectomy followed by autologous islet transplantation is a therapeutic option for the treatment of refractory chronic pancreatitis (CP). Maximization of islet yields from fibrotic and inflamed organs is crucial for prevention of posttransplant diabetes. We adapted technical advancements developed for islet allotransplantation toward islet autotransplantation. Eight patients (two men, six women; ages 24-58 years) underwent total (n = 7) or partial (n = 1) pancreatectomy for the treatment of CP refractory to maximal medical management. Pancreata were preserved in UW solution (UW group) in initial three cases and the last five pancreata were preserved with pancreatic ductal injection followed by ET-Kyoto/oxygenated PFC solutions (DI+TLM group). Islets were isolated by modified Ricordi method and were purified only in one case. All islet infusions were performed under general anesthesia via direct vein injection into the portal venous system with pressure monitoring. Total islet yields (129,314 ± 51,627 vs. 572,841 ± 116,934 IEQ, p < 0.04), islet yield/pancreas weight (1,233 ± 359 vs. 6,848 ± 847 IEQ/g, p < 0.003), and islet yield/patient body weight (1,951 ± 762 vs. 7,305 ± 1,531 IEQ/kg, p < 0.05) were significantly higher in the DI+TLM group when compared to the UW group. Pellet size was also higher (5.3 ± 0.3 vs. 13.5 ± 3.4 ml) in the DI+TLM group, suggesting that this method of preservation effectively protected pancreatic tissue against autolysis. First month posttransplant basal C-peptide and the secretory unit of islet transplant objects (SUITO) index were also higher in the DI+TLM group when compared to the UW group (2.0 ± 0.3 vs. 1.4 ± 0.4 ng/ml and 42.6 ± 12.7 vs. 14.6 ± 5.6, respectively). There were no technical complications related to the infusion. Our results suggest that higher islet yields can be achieved even from chronically inflamed and fibrotic organs using DI+TLM. The techniques applied for islet isolations from normal pancreata are showing promise for fibrotic pancreata from CP patients.
  Cell Transplantation 01/2012; 21(2-3):553-8. · 5.13 Impact Factor
• Article: Improvement of porcine islet isolation by inhibition of trypsin activity during pancreas preservation and digestion using α1-antitrypsin.

  Masayuki Shimoda, Hirofumi Noguchi, Yasutaka Fujita, Morihito Takita, Tetsuya Ikemoto, Daisuke Chujo, Bashoo Naziruddin, Marlon F Levy, Naoya Kobayashi, Paul A Grayburn, Shinichi Matsumoto
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  ABSTRACT: Porcine islets are considered to be a promising resource for xenotransplantation. However, it is difficult to isolate porcine islets because of the marked fragility and rapid dissociation. Endogenous trypsin is one of the main factors to damage islets during the isolation procedure. Recent studies have suggested that trypsin inhibitors during the preservation of pancreas or the collagenase digestion can improve the result of islet isolation. In this study, we examined whether α1-antitrypsin (Aralast™), which inhibits several endogenous proteases and has immunomodulatory properties, can protect islets from the proteases and improve the results of porcine islet isolation. Twelve porcine pancreata were divided into three groups: without Aralast group (standard, n = 5), preserved with Aralast using the ductal injection (DI) method (DI, n = 3), and with Aralast using the DI method and in the collagenase solution (DI+C, n = 4). Efficacy of islet isolation was assessed by islet yields, purity, and viability. The trypsin activity of the preservation and the digestion solution during the isolation procedure was measured. During islet isolation, the trypsin activity in DI+C group was significantly inhibited compared to the standard group, whereas DI group showed less effect than DI+C group. The average of postpurification islet equivalents (IEQ) per pancreas weight in the DI+C group was significantly higher than the standard group (standard: 3516 ± 497 IEQ/g, DI: 4607 ± 1090 IEQ/g, DI+C: 7097 ± 995 IEQ/g; p = 0.017 between standard and DI+C). In the DI+C group, stimulation index was higher than in other groups, although there was no significant difference. The presence of Aralast in both DI solution and collagenase solution markedly inhibited trypsin activity during pancreas digestion procedure and improved the porcine islet isolation. Inhibition of trypsin activity by Aralast could improve porcine islet isolation.
  Cell Transplantation 01/2012; 21(2-3):465-71. · 5.13 Impact Factor
• Article: Evaluation of osmolality of density gradient for human islet purification.

  Hirofumi Noguchi, Bashoo Naziruddin, Masayuki Shimoda, Yasutaka Fujita, Daisuke Chujo, Morihito Takita, Han Peng, Koji Sugimoto, Takeshi Itoh, Naoya Kobayashi, Nicholas Onaca, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: For pancreatic islet transplantation, the most common method of islet purification is density gradient centrifugation because of the differences in density between islets and acinar tissue. The density of islets/acinar tissue depends on several conditions, such as osmolality of purification solution. In this study, we evaluated the osmolality of iodixanol-controlled density gradients (400, 450, and 500 mOsm/kg) on the islet purification step. The density of the purification solutions was controlled by changing the volumetric ratio of iodixanol and the purification solutions (iodixanol-Kyoto solutions; IK solutions). The osmolality of density gradients was controlled by addition of 10× Hanks balanced salt solution (HBSS) solution. Density of both islets and acinar tissue increased relative to increase of the osmolality of purification solutions. There were no significant differences among the three groups on islet yield after density-adjusted purification and the rate of postpurification recovery. In vitro and in vivo assays suggest that the quality of islets was similar among the three groups. Our data suggest that efficacy of purification and quality of isolated islets is similar when the osmolality of purification solutions is between 400 and 500 mOsm/kg and density adjustment is applied. Since the density of islet and acinar tissue is changed according to osmolality, the density adjustment is important when using several osmolality solutions.
  Cell Transplantation 01/2012; 21(2-3):493-500. · 5.13 Impact Factor
• Article: Comparison of ulinastatin, gabexate mesilate, and nafamostat mesilate in preservation solution for islet isolation.

  Hirofumi Noguchi, Bashoo Naziruddin, Andrew Jackson, Masayuki Shimoda, Yasutaka Fujita, Daisuke Chujo, Morihito Takita, Han Peng, Koji Sugimoto, Takeshi Itoh, Naoya Kobayashi, Michiko Ueda, Teru Okitsu, Yasuhiro Iwanaga, Hideo Nagata, Xiaoling Liu, Hiroki Kamiya, Nicholas Onaca, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: For islet transplantation, maintaining organ viability after pancreas procurement is critically important for optimal graft function and survival. We recently reported that islet yield was significantly higher in the modified ET-Kyoto (MK) solution, which includes a trypsin inhibitor (ulinastatin), compared with the UW solution, and that the advantages of MK solution are trypsin inhibition and less collagenase inhibition. In this study, we compared ulinastatin with other trypsin inhibitors, gabexate mesilate, and nafamostat mesilate, in preservation solution for islet isolation. Ulinastatin was easily dissolved in ET-Kyoto solution, while ET-Kyoto with gabexate mesilate and nafamostat mesilate became cloudy immediately after addition. Although there were no significant differences in islet yield among the three groups, viability was significantly higher for the MK group than for the GK group or the NK group. The stimulation index was significantly higher for the MK group than for the GK group. In summary, there are no other trypsin inhibitors that are more effective than ulinastatin. Based on these data, we now use ET-Kyoto solution with ulinastatin for clinical islet transplantation.
  Cell Transplantation 01/2012; 21(2-3):509-16. · 5.13 Impact Factor
• Article: Fresh islets are more effective for islet transplantation than cultured islets.

  Hirofumi Noguchi, Bashoo Naziruddin, Andrew Jackson, Masayuki Shimoda, Tetsuya Ikemoto, Yasutaka Fujita, Daisuke Chujo, Morihito Takita, Han Peng, Koji Sugimoto, Takeshi Itoh, Naoya Kobayashi, Nicholas Onaca, Marlon F Levy, Shinichi Matsumoto
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  ABSTRACT: For clinical islet transplantation, isolated islets deteriorate rapidly in culture, although culturing islets prior to transplantation provides flexibility for evaluation of isolated islets and pretreatment of patients. In the present study, we compared human fresh islets to cultured islets with in vitro and in vivo assays. After culture for 24, 48, and 72 h, islet yield significantly decreased from 2,000 to 1,738 ± 26 (13% loss), 1,525 ± 30 (24% loss), or 1,298 ± 18 IEQ (35% loss), respectively. The ATP contents were significantly higher in the 6-h cultured group (near fresh group) than in 48-h culture groups. The stimulation index was relatively higher in the 6-h cultured group than in 48-h cultured group. Human islets with or without culture were transplanted into diabetic nude mice. The attainability of posttransplantation normoglycemia was significantly higher in fresh group than in the culture groups. Intraperitoneal glucose tolerance testing (IPGTT) showed that the blood glucose levels of mice transplanted with fresh islets were significantly lower than with cultured islets at 30, 60, 90, and 120 min after injection. These data suggest that human islet transplantation without culture could avoid the deterioration of islets during culture and improve the outcome of islet transplantation. Based on these data, we have transplanted fresh islets without culture for our current clinical islet transplantation protocol.
  Cell Transplantation 01/2012; 21(2-3):517-23. · 5.13 Impact Factor
• Article: Implications of a positive crossmatch in liver transplantation: a 20-year review.

  Richard Ruiz, Koji Tomiyama, Jeffrey Campsen, Robert M Goldstein, Marlon F Levy, Greg J McKenna, Nicholas Onaca, Brian Susskind, Glenn W Tillery, Goran B Klintmalm
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  ABSTRACT: Whether a positive crossmatch result has any relevance to liver transplantation (LT) outcomes remains controversial. We assessed the impact of a positive crossmatch result on patient and graft survival and posttransplant complications. During a 20-year period, 2723 LT procedures with crossmatch results were identified: 2479 primary transplants and 244 retransplants. The rates of positive B cell and T cell crossmatches were 10.1% and 7.4%, respectively, for primary transplants and 14.6% and 6.4%, respectively, for retransplants (P = 0.049 for a B cell crossmatch). Across all primary transplants, females (P < 0.001) and patients with autoimmune hepatitis (P < 0.001) had greater frequencies of positive crossmatches. There was no effect from race or age. For both primary transplants and retransplants, patient survival and graft survival were not affected by the presence of a positive crossmatch. With respect to posttransplant complications, there were no differences in rejection episodes (hyperacute, acute, or chronic) or technical complications (biliary and vascular) between negative and positive crossmatch groups. However, there were significant differences in the pathological findings of preservation injury (PI) on liver biopsy samples taken at the time of transplantation and within the first week of transplantation (P = 0.003 for B cells and P = 0.03 for T cells). In summary, a positive crossmatch had no significant impact on patient survival or graft outcomes. However, there was a significantly higher incidence of PI in primary LT recipients with a positive crossmatch. This finding is important for a broader understanding of PI, which may include a significant immunological component.
  Liver Transplantation 12/2011; 18(4):455-60. · 3.39 Impact Factor