|ognizant Communication Corporation|
The Regenerative Medicine Journal
VOLUME 19, NUMBER 3, 2010
Cell Transplantation, Vol. 19, pp. 253-268, 2010
0963-6897/10 $90.00 + 00
Copyright © 2010 Cognizant Comm. Corp.
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Immune Tolerance Induction by Integrating Innate and Adaptive Immune Regulators
Jun Suzuki,1 Camillo Ricordi,1,2,3,4 and Zhibin Chen1,2
1Department of Microbiology and Immunology, University of
Miami, Miami, FL, USA
2Diabetes Research Institute, University of Miami, Miami, FL, USA
3Department of Surgery, University of Miami, Miami, FL, USA
4Karolinska Institute, Stockholm, Sweden
A diversity of immune tolerance mechanisms have evolved to protect normal tissues from immune damage. Immune regulatory cells are critical contributors to peripheral tolerance. These regulatory cells, exemplified by the CD4s+Foxp3+ regulatory T (Treg) cells and a recently identified population named myeloid-derived suppressor cells (MDSCs), regulate immune responses and limiting immune-mediated pathology. In a chronic inflammatory setting, such as allograft-directed immunity, there may be a dynamic "cross-talk" between the innate and adaptive immunomodulatory mechanisms for an integrated control of immune damage. CTLA4-B7-based interaction between the two branches may function as a molecular "bridge" to facilitate such "cross-talk." Understanding the interplays among Treg cells, innate suppressors, and pathogenic effector T (Teff) cells will be critical in the future to assist in the development of therapeutic strategies to enhance and synergize physiological immunosuppressive elements in the innate and adaptive immune system. Successful development of localized strategies of regulatory cell therapies could circumvent the requirement for very high number of cells and decrease the risks associated with systemic immunosuppression. To realize the potential of innate and adaptive immune regulators for the still elusive goal of immune tolerance induction, adoptive cell therapies may also need to be coupled with agents enhancing endogenous tolerance mechanisms.
Key words: Immune tolerance; Immunosuppression; Regulatory cells; CTLA4; Autoimmunity; Transplant
Address correspondence to Zhibin Chen, M.D., Ph.D., Department of Microbiology and Immunology, University of Miami, RMSB 3035,1600 NW 10th Ave, Miami, FL 33136, USA. Tel: 305-243-8348; Fax: 305-243-4404; E-mail: firstname.lastname@example.org
Continuous and High-Level In Vivo Delivery of Endostatin From Recombinant Cells Encapsulated in TheraCyte® Immunoisolation Devices
N. V. Malavasi,1 D. B. Rodrigues,1 R. Chammas,2 R. M. Chura-Chambi,1 J. A. M. Barbuto,3 K. Balduino,1 S. Nonogaki,4 and L. Morganti1
1Instituto de Pesquisas Energéticas e Nucleares-IPEN-CNEN/SP,
Centro de Biotecnologia, São Paulo, Brazil
2Laboratório de Oncologia Experimental, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil
3Departamento de Imunologia do Instituto de Ciências Biomédicas da USP, São Paulo, Brazil
4Instituto Adolfo Lutz, São Paulo, Brazil
Endostatin (ES) is a potent inhibitor of angiogenesis and tumor growth. Continuous ES delivery of ES improves the efficacy and potency of the antitumoral therapy. The TheraCyte® system is a polytetrafluoroethylene (PTFE) semipermeable membrane macroencapsulation system for implantation of genetically engineered cells specially designed for the in vivo delivery of therapeutic proteins, such as ES, which circumvents the problem of limited half-life and variation in circulating levels. In order to enable neovascularization at the tissues adjacent to the devices prior to ES secretion by the cells inside them, we designed a scheme in which empty TheraCyte® devices were preimplanted SC into immunodeficient mice. Only after healing (17 days later) were Chinese hamster ovary cells expressing ES injected into the preimplanted devices. In another model for device implantation, the cells expressing ES where loaded into the immunoisolation devices prior to implantation into the animals, and the TheraCyte® were then immediately implanted SC into the mice. Throughout the 2-month study, constant high ES levels of up to 3.7 mg/ml were detected in the plasma of the mice preimplanted with the devices, while lower but also constant levels of ES (up to 2.1 mg/ml plasma) were detected in the mice that had received devices preloaded with the ES-expressing cells. Immunohistochemistry using anti-ES antibody showed reaction within the device and outside it, demonstrating that ES, secreted by the confined recombinant cells, permeated through the membrane and reached the surrounding tissues.
Key words: Endostatin; Immunoisolation; Angiogenesis; Gene therapy
Address correspondence to Ligia Morganti, Instituto de Pesquisas Energéticas e Nucleares-IPEN-CNEN/SP, Centro de Biotecnologia, Av. Prof. Lineu Prestes, 2242, São Paulo, Brazil. Tel: 55-11-3133-9703; Fax: 55-11-3133-9709; E-mail: email@example.com
Isolation, Characterization, and Differentiation Potential of Canine Adipose-Derived Stem Cells
N. M. Vieira,1 V. Brandalise,1 E. Zucconi,1 M. Secco,1 B. E. Strauss,2 and M. Zatz1
1Human Genome Research Center, Biosciences Institute, University
of São Paulo, São Paulo, Brazil
2Viral Vector Group, Heart Institute, InCor, University of São Paulo, São Paulo, Brazil
Adipose tissue may represent a potential source of adult stem cells for tissue engineering applications in veterinary medicine. It can be obtained in large quantities, under local anesthesia, and with minimal discomfort. In this study, canine adipose tissue was obtained by biopsy from subcutaneous adipose tissue or by suction-assisted lipectomy (i.e., liposuction). Adipose tissue was processed to obtain a fibroblast-like population of cells similar to human adipose-derived stem cells (hASCs). These canine adipose-derived stem cells (cASCs) can be maintained in vitro for extended periods with stable population doubling and low levels of senescence. Immunofluorescence and flow cytometry show that the majority of cASCs are of mesodermal or mesenchymal origin. cASCs are able to differentiate in vitro into adipogenic, chondrogenic, myogenic, and osteogenic cells in the presence of lineage-specific induction factors. In conclusion, like human lipoaspirate, canine adipose tissue may also contain multipotent cells and represent an important stem cell source both for veterinary cell therapy as well as preclinical studies.
Key words: Canine adipose-derived stem cells; Tissue engineering; Veterinary cell therapy
Address correspondence to Dr. Mayana Zatz, Ph.D., Human Genome Research Center, Institute of Biosciences, University of São Paulo, Rua do Matão, n.106 Cidade Universitária, São Paulo-SP, Brasil-CEP: 05508-090. Tel/Fax: (55) (11) 3091-7966; E-mail: firstname.lastname@example.org
Seven Consecutive Successful Clinical Islet Isolations With Pancreatic Ductal Injection
Shinichi Matsumoto,1* Hirofumi Noguichi,1* Masayuki Shimoda,2 Tetsuya Ikemoto,1 Bashoo Naziruddin,2 Andrew Jackson,,2,3 Yoshiko Tamura,2 Greg Olson,2 Yasutaka Fujita,1 Daisuke Chujo,4 Morihito Takita,1 Naoya Kobayashi,5 Nicholas Onaca,2 and Marlon Levy1,2
1Baylor Research Institute, Fort Worth, TX, USA
2Baylor University Medical Center, Dallas, TX, USA
3Biomedical Studies, Baylor University, Waco, TX, USA
4Baylor Institute for Immunology Research, Dallas, TX, USA
5Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
Inconsistent islet isolation is one of the issues of clinical islet transplantation. In the current study, we applied ductal injection to improve the consistency of islet isolation. Seven islet isolations were performed with the ductal injection of ET-Kyoto solution (DI group) and eight islet isolations were performed without the ductal injection (standard group) using brain-dead donor pancreata. Isolated islets were evaluated based on the Edmonton protocol for transplantation. The DI group had significantly higher islet yields (588,566 ± 64,319 vs. 354,836 ± 89,649 IE, p < 0.01) and viability (97.3 ± 1.2% vs. 92.6 ± 1.2%, p < 0.02) compared with the standard group. All seven isolated islet preparations in the DI group (100%), versus only three out of eight isolated islet preparations (38%) in the standard group met transplantation criteria. The islets from the DI group were transplanted into three type 1 diabetic patients and all three patients became insulin independent. Ductal injection significantly improved quantity and quality of isolated islets and resulted in high success rate of clinical islet transplantation. This simple modification will reduce the risk of failure of clinical islet isolation.
Key words: Ductal injection; Islet isolation; Islet transplantation; Brain-dead donor; ET-Kyoto solution
Address correspondence to Shinichi Matsumoto, M.D., Ph.D., Baylor All Saints Medical Center/Baylor Research Institute, 1400 8th Avenue, Fort Worth, TX 76104, USA. Tel: 817-922-2570; Fax: 817-922-4645; E-mail: email@example.com
*Shinichi Matsumoto and Hirofumi Noguchi contributed equally to this work.
Enhanced Prediction of Porcine Islet Yield and Posttransplant Outcome Using a Combination of Quantitative Histomorphometric Parameters and Flow Cytometry
Sang-Man Jin,1,2,3 Kang Seok Kim,1,2,3 Song-Yi Lee,1,2,3 Chang-Hoon Gong,1,2,3 Su Kyoung Park,1,2,3 Jae Eun Yu,1,2 Su-Cheong Yeom,1,4 Tai Wook Yoon,1,2 Jongwon Ha,1,6,7 Chung-Gyu Park,1,2,3,5,6,7 and Sang-Joon Kim1,6,7
1Xenotransplantation Research Center, Seoul National University
Hospital, Seoul, Republic of Korea
2Cancer Research Institute, Seoul National University College of Medicine, Seoul, Republic of Korea
3Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
4Center for Animal Resource Development, Seoul National University College of Medicine, Seoul, Republic of Korea
5Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, Republic of Korea
6Transplantation Research Institute, Seoul National University College of Medicine, Seoul, Republic of Korea
7Department of Surgery, Seoul National University College of Medicine, Seoul, Republic of Korea
Prediction of islet yield and posttransplant outcome is essential for clinical porcine islet xenotransplantation. Although several histomorphometric parameters of biopsied porcine pancreases are predictive of islet yield, their role in the prediction of in vivo islet potency is unknown. We investigated which histomorphometrical parameter best predicts islet yield and function, and determined whether it enhanced the predictive value of in vitro islet function tests for the prediction of posttransplant outcome. We analyzed the histomorphometry of pancreases from which 60 adult pig islet isolations were obtained. Islet function was assessed using the b-cell viability index based on flow cytometry analysis, oxygen consumption rate, ADP/ATP ratio, and/or concurrent transplantation into NOD/SCID mice. Receiver operating characteristic (ROC) analysis revealed that only islet equivalent (IEQ)/cm2 and the number of islets >200 mm in diameter significantly predicted an islet yield of >2000 IEQ/g (p < 0.001 for both) and in vivo islet potency (p = 0.024 and p = 0.019, respectively). Although not predictive of islet yield, a high proportion of large islets (>100 mm in diameter) best predicted diabetes reversal (p = 0.001). Multiple regression analysis revealed that the b-cell viability index (p = 0.003) and the proportion of islets >100 mm in diameter (p = 0.048) independently predicted mean posttransplant blood glucose level (BGL). When BGL was estimated using both these parameters [area under the ROC curve (AUC), 0.868; 95% confidence interval (CI), 0.730-1.006], it predicted posttransplant outcome more accurately than the b-cell viability index alone (AUC, 0.742; 95% CI, 0.544-0.939). In conclusion, we identified the best histomorphometric predictors of islet yield and posttransplant outcome. This further enhanced the predictive value of the flow cytometry analysis. These parameters should be useful for predicting islet yield and in vivo potency before clinical adult porcine islet xenotransplantation.
Key words: Biopsy; Pancreas; Flow cytometry; Islet isolation; Islet potency
Address correspondence to Chung-Gyu Park, M.D., Ph.D., Department of Microbiology and Immunology, Xenotransplantation Research Center, Seoul National University College of Medicine, 103 Daehangno Jongno-gu, Seoul 110-799, Republic of Korea. Tel: +82-2-740-8308; Fax: +82-2-743-0881; E-mail: firstname.lastname@example.org
Transplantation of Mesenchymal Stem Cells Exerts a Greater Long-Term Effect Than Bone Marrow Mononuclear Cells in a Chronic Myocardial Infarction Model in Rat
Manuel Mazo,1 Juan José Gavira,2 Gloria Abizanda,1 Cristina Moreno,3 Margarita Ecay,4 Mario Soriano,6 Pablo Aranda,1 María Collantes,4 Eduardo Alegría,2 Juana Merino,3 Iván Peñuelas,5 José Manuel García Verdugo,6 Beatriz Pelacho,1* and Felipe Prósper1*
1Hematology and Cell Therapy and Division of Cancer, Clinica
Universitaria and Foundation for Applied Medical Research, University of
Navarra, Navarra, Spain
2Department of Cardiology and Cardiovascular Surgery, University of Navarra, Navarra, Spain
3Immunology Service, University of Navarra, Navarra, Spain
4MicroPET Research Unit CIMA-CUN, University of Navarra, Navarra, Spain
5Radiopharmacy Unit, Department of Nuclear Medicine Clínica Universitaria, University of Navarra, Navarra, Spain
6Department of Cell Biology, Instituto Cavanilles, University of Valencia, Valencia, Spain
The aim of this study is to assess the long-term effect of mesenchymal stem cells (MSC) transplantation in a rat model of chronic myocardial infarction (MI) in comparison with the effect of bone marrow mononuclear cells (BM-MNC) transplant. Five weeks after induction of MI, rats were allocated to receive intramyocardial injection of 106 GFP-expressing cells (BM-MNC or MSC) or medium as control. Heart function (echocardiography and 18F-FDG-microPET) and histological studies were performed 3 months after transplantation and cell fate was analyzed along the experiment (1 and 2 weeks and 1 and 3 months). The main findings of this study were that both BM-derived populations, BM-MNC and MSC, induced a long-lasting (3 months) improvement in LVEF (BM-MNC: 26.61 ± 2.01% to 46.61 ± 3.7%, p < 0.05; MSC: 27.5 ± 1.28% to 38.8 ± 3.2%, p < 0.05) but remarkably, only MSC improved tissue metabolism quantified by 18FFDG uptake (71.15 ± 1.27 to 76.31 ± 1.11, p < 0.01), which was thereby associated with a smaller infarct size and scar collagen content and also with a higher revascularization degree. Altogether, results show that MSC provides a long-term superior benefit than whole BM-MNC transplantation in a rat model of chronic MI.
Key words: Bone marrow stem cells; Chronic myocardial infarction; Cardiac remodeling; Angiogenesis
Address correspondence to Felipe Prósper, M.D., Hematology and Cell Therapy, Clínica Universitaria,31008, Navarra, Spain. Tel: 34 948 194700; E-mail: email@example.com
*B.P. and F.P. contributed equally to this work and should be considered equal last authors.