Vascularized Composite Allotransplantation最新文献

{"title":"2586: Next generation quantitative sequencing for detecting chimerism in a NHP model","authors":"J. Woodall, M. Uluer, Matthew T. Chrencik, A. Nam, S. Bartlett, R. Barth","doi":"10.1080/23723505.2016.1234247","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234247","url":null,"abstract":"2586: Next generation quantitative sequencing for detecting chimerism in a NHP model Jhade D. Woodall, MD, Mehmet C. Uluer, MD, ScM, Matthew Chrencik, Arthur J. Nam, MD, Stephen T. Bartlett, MD, and Rolf N. Barth, MD University of Maryland Medical Center; University of Maryland School of Medicine, Baltimore, MD, USA Background The association between chimerism and immunologic tolerance in transplantation has shown a path toward eliminating the need for lifelong immunosuppression We have been investigating vascularized composite allograft (VCA) facial transplantation in a well-established non-human primate (NHP) model and have observed transient chimerism without the use of preconditioning therapies when these grafts include vascularized bone marrow The detection of chimerism in the Mauritian cynomolgus macaque (MCM) model relies on flow cytometry with limited ability to detect microchimerism We explored the use of next generation quantitative sequencing as a method of chimerism detection in our VCA model. Methods: Non-specific PCR primers were designed in a conserved region to amplify all Mafa-B major alleles for each of the 7 haplotypes (M1-M7) in MCMs Two animals DNA were initially used: with M3/M3 and M6/M6 haplotypes After confirming PCR efficiency and that products were of appropriate length the primers were coupled with barcodes for use with the ion torrent sequencing system The barcoded primers were added to mixed DNA from M3 and M6 animals and amplified for 25 cycles These products were purified and then sequenced by ion torrent. Results Initial efficiency was determined with the non-barcoded primers on 2 different animals (M3/M3, M6/ M6) to ensure our conditions and primers were at an optimal range These haplotypes had efficiencies in a reasonable range between 90–105% The addition of the barcode to these primers resulted in a drop in the PCR efficiencies to 77% and 79% respectively both with R values of 099 Despite similar but decreased efficiencies, sequencing was subsequently performed Sequencing results indicated that both haplotypes were present at inexact ratios, thus not permitting quantitative assessments of chimerism. Conclusions Real time PCR and next generation quantitative sequencing technologies are feasible techniques to detect chimerism in our MCM model Optimization of PCR reaction conditions for barcoded primers are necessary for quantitative interpretations Furthermore, more specific primers for sets of haplotypes may be necessary given the genetically close nature of MCM’s CONTACT Mehmet C. Uluer, MD, ScM mculuer@gmail.com © 2016 Jhade D. Woodall, Mehmet C. Uluer, Matthew Chrencik, Arthur J. Nam, Stephen T. Bartlett, and Rolf N. Barth. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"37 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123542420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2542: Exosomes from mesenchymal stem cells package anti-inflammatory cytokines in allotransplantation","authors":"Jonathan P. Massie, Yohei Rosen, J. Diaz-Siso, N. Plana, Daniel J Ceradini","doi":"10.1080/23723505.2016.1232965","DOIUrl":"https://doi.org/10.1080/23723505.2016.1232965","url":null,"abstract":"2542: Exosomes from mesenchymal stem cells package anti-inflammatory cytokines in allotransplantation Jonathan P. Massie, BS, Yohei M. Rosen, J. Rodrigo Diaz-Siso, Natalie M. Plana, and Daniel J. Ceradini NYU Langone Medical Center, New York, NY, USA","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123822048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2576: Optimization of a differential cytokine profile-based non-invasive diagnostic and predictive tool for reliable diagnosis of acute rejection in VCA","authors":"Piul S. Rabbani, Rohini L Kadle, Nakul Rao, Chin Park, Daniel J Ceradini","doi":"10.1080/23723505.2016.1233019","DOIUrl":"https://doi.org/10.1080/23723505.2016.1233019","url":null,"abstract":"2576: Optimization of a differential cytokine profile-based non-invasive diagnostic and predictive tool for reliable diagnosis of acute rejection in VCA Piul S. Rabbani, PhD, Rohini L. Kadle, MD, Nakul Rao, Chin Park, and Daniel Ceradini New York University School of Medicine, New York, NY, USA Background Current methods of detection of early acute transplant rejection relies on invasive tissue biopsies and time-consuming histological analysis We propose an alternative method using adhesive discs to analyze molecular changes in cells sampled from the epidermis of a vascular composite allotransplant (VCA), to detect markers of acute rejection We aim to validate efficiency of skin stripping as a non-invasive predictor and sensitive diagnostic test for acute rejection in VCA. Methods Using an established VCA rat model, we transplanted composite flaps from donor Brown-Norway rats to age-matched recipient Lewis rats Following cyclosporine for 5 days, we inspected daily for clinical signs of rejection and sampled transplanted skin with adhesive CuDerm-discs at each time point up to rejection We performed QRT-PCR on sampled cells for cytokines associated with early rejection We sampled flaps for biopsies and histology to corroborate the disc data. Results CuDerm-disc-sourced PCR revealed that expressions of MCP1, MIP1̂I§, MIP1̂I and CXCL10 increased progressively with mild and advanced rejection, compared to the immunosuppressed stage (p < 005) MIP3̂I§ and CXCL9 showed significant upregulation at mild rejection (19-fold, 70-fold, respectively), but a downregulation during advanced rejection (4fold,20-fold, respectively, p < 005) Comparison of mild and advanced rejection showed highly significant differential cytokine expression (p < 001) We verified the sensitivity and validity of the CuDerm-disc method by comparison of mRNA expression in VCA biopsies and found cytokine detection comparable between both methods The mild and advanced rejection cytokine profiles from discs also corresponded with the Banff classification of cellular allograft rejection of the respective flap histologies. Conclusions Skin stripping, when compared to traditional tissue biopsy, is a comparable and reliable analytical tool The cytokine profiles gathered from skin stripping are distinct and capable of detecting the earliest stages of acute rejection, as well as distinguishing from advanced rejection, stages which are difficult to analyze definitively using traditional histology Our results clearly demonstrate the promise of skin stripping as a noninvasive tool in predicting and diagnosing early rejection, prior to onset of advanced rejection.","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131692629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2510: Elbow vascularized composite allotransplantation - Surgical anatomy and technique","authors":"Zvi Steinberger, Heng Xu, N. Kazmers, S. Thibaudeau, Russell G. Huffman, L. Levin","doi":"10.1080/23723505.2016.1234219","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234219","url":null,"abstract":"2510: Elbow vascularized composite allotransplantation Surgical anatomy and technique Zvi Steinberger, MD, Heng Xu, Nikolas H. Kazmers, Stephanie Thibaudeau, Russel G. Huffman, and L. Scott Levin University of Pennsylvania, Philadelphia, PA, USA Background Elbow reconstruction with vascularized composite allotransplantation (VCA) may hold promise in treating end-stage arthritis, as no current treatment is both functional and durable. We describe the vascular and gross anatomy of the elbow in the context of VCA procurement, and propose a step-by-step surgical technique for elbow VCA. Methods Sixteen fresh adult cadaveric upper extremities underwent arterial tree latex injection. Arteries, nerves, and their branch points were identified and measured relative to the medial epicondyle. Based upon our determination of the dominant blood supply to elbow osseous and capsular structures, a cadaveric model of elbow VCA was derived by performing donor preparation on two fresh cadaveric upper extremities, with elevation of a lateral arm flap in conjunction with the vascularized elbow joint. Two size-matched specimens underwent recipient preparation, followed by transplantation. The surgical technique was refined with each successive transplant. Results The arterial supply to the elbow was comprised of consistent branches contributing to medial, lateral, and posterior arcades (Table 1). Preservation of the elbow arterial network requires sectioning of brachial, radial and ulnar arteries 12 cm proximal, 1 cm distal, and 6 cm distal to the ulnar artery take-off, respectively. Preservation of the supinator, anconeus, distal brachialis, proximal aspects of the flexor digitorum profundus, and flexor carpi ulnaris is required to protect osseous perforators. Nerves branches to the joint most commonly were derived from ulnar and median nerves (Table 1). Following two cadaveric elbow VCA procedures, our proposed surgical technique has been refined (Table 2).","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"15 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128226612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2599: A volumetric analysis of potential tissue burden for lower extremity vascularized composite allotransplantation","authors":"M. Carty, S. Talbot, S. Ledbetter, Seth Levine","doi":"10.1080/23723505.2016.1234264","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234264","url":null,"abstract":"2599: A volumetric analysis of potential tissue burden for lower extremity vascularized composite allotransplantation Matthew J. Carty, MD, Simon G. Talbot, MD, Stephen Ledbetter, MD, MPH, and Seth Levine Brigham and Women’s Hospital, Boston, MA, USA; Brigham and Women’s Faulkner Hospital, Boston, MA, USA Background Despite remarkable advances in vascularized composite allotransplantation (VCA) over the past 2 decades, there has been general resistance to embrace the notion of lower extremity allotransplantation One of the principal concerns raised is the potential tissue burden associated with lower extremity allotransplantation; however, no formal studies to assess the anticipated volume of biological substrate to be transferred in lower extremity VCA has been performed to date. Methods We performed volumetric analyses of 10 patients’ lower extremities utilizing DICOM data from standard lower extremity CT scans and Vitrea Enterprise Suite 674 software For each patient, standard 3D renderings of the lower extremities were generated and segmented into discrete tissue phases based on Hounsfield units Volumetric assessment were then made of each lower extremity at both the below knee (BK) level and above knee (AK) level Similar volumetric analyses were performed on a single patient’s upper extremity CT scan to use as reference. Results Ten patients’ lower extremity CT scans were analyzed. Mean patient age was 52§7 years, with mean BMI of 29 § 4 and a 50:50 male:female gender split Volumetric analyses at the BK level were: total 2660 § 676cc, muscle 1369 § 307cc, fat 859 § 387, bone 432 § 79cc; at the AK level: total 7483 § 1731cc, muscle 3632 § 768cc, fat 2940 § 1203cc, bone 921 § 159cc By comparison, a full upper extremity control demonstrated the following volumetric results: total 2968cc, muscle 2005cc, fat 629cc, bone 334cc Approximate matching of a subset of the lower extremity patient analyses to the upper extremity control revealed that a unilateral BK transplant would involve transfer of 90% of the total tissue volume of a full upper extremity, while a unilateral AK transplant would involve transfer of 252% of the total tissue volume of the same. Conclusions In the case of BK level transplants, anticipated tissue transfer will likely be less than that already performed successfully in high upper extremity transplant procedures AK level transplants, however, will entail a higher level of tissue burden than what has previously been witnessed in VCA cases reported worldwide, and may require special considerations to mitigate the likely metabolic impact of larger scale ischemia-reperfusion injury and immunologic load. CONTACT Matthew J. Carty, MD mcarty@partners.org © 2016 Matthew J. Carty, Simon G. Talbot, Stephen Ledbetter, and Seth Levine. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130980110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2601: Genitourinary vascularized composite allotransplanation: Preliminary report of first case performed in the United States","authors":"C. Cetrulo, B. Bojovic, K. Eberlin, J. Winograd, F. Mcgovern, Michael P. Grant, C. Tanrikut, A. Feldman, R. Ehrlichman, H. Salinas, M. Treiser, Jeffrey Lee, E. Wright, K. Lee, Ilse M. Schol, Paul W Holzer, W. Austen, D. Ko","doi":"10.1080/23723505.2016.1234212","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234212","url":null,"abstract":"2601: Genitourinary vascularized composite allotransplanation: Preliminary report of first case performed in the United States Curtis L. Cetrulo, MD, FACS, Branko Bojovic, MD, Kyle Eberlin, MD, Jonathan Winograd, MD, Francis McGovern, MD, Michael Grant, MD, Cigdem Tanrikut, MD, Adam Feldman, MD, Richard Ehrlichman, MD, Harry Salinas, MD, Matthew Treiser, MD, Jeffrey Lee, MD, Eric Wright, MD, Kai Lee, MD, Ilse Schol, BS, Paul Holzer, MS, William G. Austen, Jr., MD, and Dicken Ko, MD Massachusetts General Hospital, Boston, MA, USA Introduction Vascularized Composite Allotransplantation (VCA) has been a clinical reality in hand transplantation since the late 1990s and in facial transplantation since 2005. VCA has ushered in a very exciting and explosive era of research and new modalities to offer patients in need of options the potential for restorative operative interventions. In addition to upper extremity and facial VCA, much interest has been given to genitourinary VCA, specifically penile transplantation. Recent conflicts that have rendered many wounded warriors with devastating genitourinary injuries, a subset of penile cancer patient survivors previously left with disfiguring or minimal residual penile function, and a newer group of transgender patients interested in more realistic reconstruction than current autologous stateof-the-art techniques can offer, have furthered research and interest in penile VCA. Two prior successful penile VCA cases have been performed to date in the world literature experience. Methods A protocol for penile VCA was approved by the Internal Review Board at Massachusetts General Hospital (MGH) Appropriate candidate screening and selection was approved by the patient selection committee. Preoperative preparation via recipient patient defect-specific cadaveric dissection sessions was performed. The New England Organ Bank (NEOB) facilitated donor patient screening for protocol screened and approved recipient candidates. Results In early May, 2016, a potential donor was identified by the NEOB for a recipient patient accepted by the IRB-approved protocol at MGH. Team mobilization ensued and the recipient patient was informed. Subsequently, the first successful penile VCA in the United States was performed over the course of a 15 hour operation. One week out from the successful procedure, the recipient patient is doing well and meeting expected milestones in recovery. Conclusions We herein present the first successful penile VCA performed in the United States and have demonstrated proof of concept. This now becomes the third case in the world literature experience and opens the discussion of genitourinary VCA to a new geographic focus in the world, possibly leading to further such transplants to now be considered and performed in this important and deserving patient population. CONTACT Curtis L. Cetrulo ccetrulo@mgh.harvard.edu © 2016 Curtis L. Cetrulo, Branko Bojovic, Kyle Eberlin, Jonathan Winograd, Francis McGovern, ","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"48 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132360477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2500: Possible accommodation in a bilateral transhumeral transplant [sessiontypeP]","authors":"P. Cavadas, A. Thione","doi":"10.1080/23723505.2016.1234217","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234217","url":null,"abstract":"2500: Possible accommodation in a bilateral transhumeral transplant [sessiontypeP] Pedro C. Cavadas, MD, PhD and Alessandro Thione, MD, PhD Clinica Cavadas, Valencia, Spain Immunological accommodation is a described intriguing mechanism by which DSA are present in a transplanted patient without evidence of organ damage. It has been described in solid organ transplantations but, to the best of the authors knowledge, not in VCA. A young male patient with bilateral transhumeral amputation was transplanted in 2008. The clinical and immunological evolution was relatively uneventful for 4 years, with excellent function. In 2012 the patient tested positive for DSA (anti DQ7, DQ8, DQ9) complement-fixing, at high MFI (16000), without clinical or histological evidence of rejection. The patient was treated with Bortezomib, 4 cycles, without clearance of the DSA. The MFI decreased to around 4000 and have remained like this ever since. Given the complete absence of clinical or pathological evidence of rejection, and the risk of complications with escalating depleting treatments, the decision of not giving further treatment was taken. Four years after the first positive DSA test, the clinical and pathological findings are normal, without evidence of organ damage. If this case constitutes a true accommodation or just an inconspicuous impending rejection will require longer follow-up. CONTACT Pedro C. Cavadas, MD, PhD pcavadas@telefonica.net © 2016 Pedro C. Cavadas and Alessandro Thione. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. VASCULARIZED COMPOSITE ALLOTRANSPLANTATION 2016, VOL. 3, NOS. 1–2, 34 http://dx.doi.org/10.1080/23723505.2016.1234217","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128860332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2527: The International Registry on Hand and Composite Tissue Allotransplantation (IRHCTT)","authors":"P. Petruzzo, J. Dubernard, M. Lanzetta","doi":"10.1080/23723505.2016.1232944","DOIUrl":"https://doi.org/10.1080/23723505.2016.1232944","url":null,"abstract":"2527: The International Registry on Hand and Composite Tissue Allotransplantation (IRHCTT) Palmina Petruzzo, MD, Jean Michel Dubernard, and Marco Lanzetta, MD Hopital Edouard Herriot, Lyon, France; Italian Institute of Hand Surgery, Monza, Italy Background The primary purpose of the IRHCTT is to collect information on voluntary basis. At present it includes upper extremity (UET) and face allotransplantations (FT). Methods 25 unilateral and 31 bilateral UET, for a total of 56 patients have been reported. In the majority of cases the level of amputation was distal, but there were also 9 arm transplantations. Twenty-eight cases of partial or total face allotransplantations have been reported. In the majority of cases the deficit included cheek, nose, chin, lips and perioral area. In both types of transplantation the immunosuppressive therapy included tacrolimus, mycophenolate mofetil, sirolimus and steroids; polyclonal or monoclonal antibodies were used for induction. Results Patient survival in UET was 9637%: 1 patient died after simultaneous face and bilateral hand transplantation and another one after bilateral arm transplantation; while in FT it was 8572% (4 patients died including the case of simultaneous face and bilateral hand transplantation) Graft survival in UET was 8182%: in 5 cases it occurred in the first period after transplantation (poor vascularization or infectious complications) and in other 5 during the follow-up (chronic rejection/graft vasculopathy) Graft survival in FT was 9643% (one face graft was removed for unknown cause). In UET 74% of the recipients experienced at least one episode of acute rejection within the first posttransplant year and 60% in FT Six cases of chronic rejection in UET and one in FT have been reported Complications included, as in solid organ transplantation, opportunistic infections, metabolic complications and malignancies. Hand-grafted patients developed protective sensibility, 90% of them tactile sensibility and 823% developed also a partial discriminative sensibility Motor recovery enabling patients to perform most daily activities Face-grafted patients improved their aesthetic aspect and they were able to perform some activities such as eating, drinking and speaking which were impossible before the transplantation. Conclusions UET and FT are successful procedures, however careful evaluation of patients before and after transplantation are indispensable. CONTACT Palmina Petruzzo, MD petruzzo@medicina.unica.it © 2016 Palmina Petruzzo, Jean Michel Dubernard, and Marco Lanzetta. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. VASCULARIZED COMPOSI","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"52 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128950771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2534: Type-1 interferon counters the immunoregulatory activity of IL-10: A mechanism in the abrogation of transplant tolerance","authors":"Marcos Iglesias Lozano, A. Arun, B. Lam, W. Lee, G. Brandacher, G. Raimondi","doi":"10.1080/23723505.2016.1232957","DOIUrl":"https://doi.org/10.1080/23723505.2016.1232957","url":null,"abstract":"2534: Type-1 interferon counters the immunoregulatory activity of IL-10: A mechanism in the abrogation of transplant tolerance Marcos Iglesias Lozano, DVM, PhD, Anirudh Arun, BS, Brandon Lam, BS, W. P. Andrew Lee, MD, Gerald Brandacher, MD, and Giorgio Raimondi, PhD Johns Hopkins University School of Medicine, Vascularized Composite Allotransplantation (VCA) Laboratory, Baltimore, MD, USA Background A growing body of evidence shows that induction of long term transplant survival by costimulation blockade (CoB) regimens is impaired by inflammatory responses In particular, multiple studies reported that engagement of toll like receptors (TLR) abrogates the tolerogenic effect of CoB Despite the identification of type-1 interferons (TI-IFN) as mediators of this effect in multiple models, the target population and specific pathway used by TI-IFN to induce this effect remain unknown To better understand how an inflammatory environment, and more specifically IFN-b, could interfere with the induction of transplant tolerance we studied their impact on the immunomodulatory properties of IL-10. Methods Mouse bulk T cells were isolated by negative-selection and Tmem and Treg subpopulations identified by flow cytometry Phospho-STAT3 induction (a key signaling step) after IL-10 and IL-6 stimulation in indicated conditions in Tmem and Treg cells were measured via flow cytometry The gene expression profile of T cell subsets exposed to TI-IFN was assessed by microarray and quantitative PCR analysis Protein levels were measured by Western Blot. Results Following 48 h of bystander incubation with IFN-b, Tmem and Treg subpopulations present a dramatic defect in the production of phospho-STAT3 in response to IL-10, but not to IL-6 Microarray and flow cytometry data indicated that this IL-10-specific unresponsiveness was not associated with any reduction of IL-10 receptor expression or an increase in SOCS (Suppressor of Cytokine Signaling) 1 and 3, nor with reduced STAT3 cytoplasmic availability They suggested a role for STAT1 in this process After IFN-b exposure, there is a complete reversal of the STAT1/ STAT3 ratio in T cells Using STAT1-KO cells, we show that the absence of STAT1 prevented/reduced IL-10 inhibition by IFN-b in Treg and Tmem, respectively. Conclusions Overall, these data reveal a promising new molecular mechanism where IFN-b interfere with IL-10 signaling in T cells via STAT1 Our data suggest that STAT1 exerts cross-competition with STAT3 for IL10R binding, preventing its phosphorylation and activity A targeted regulation of this mechanisms that counteract IL-10 suppressive functions could be a powerful tool to improve the efficacy of immunomodulatory strategies for transplant tolerance induction. CONTACT Marcos Iglesias Lozano, DVM, PhD miglesi1@jhmi.edu © 2016 Marcos Iglesias Lozano, Anirudh Arun, Brandon Lam, W. P. Andrew Lee, Gerald Brandacher, and Giorgio Raimondi. Published with license by Taylor & Francis. This is an Open Access article distr","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"46 23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123828685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"2564: Total human eye allotransplantation (THEA): Protocol optimization of imaging modalities in a non-human primate model","authors":"E. Davidson, J. Carney, B. Lopresti, M. Miller, K. Washington","doi":"10.1080/23723505.2016.1234225","DOIUrl":"https://doi.org/10.1080/23723505.2016.1234225","url":null,"abstract":"2564: Total human eye allotransplantation (THEA): Protocol optimization of imaging modalities in a non-human primate model Edward H. Davidson, MA, (Cantab), MBBS, Jonathan Carney, Brian Lopresti, Maxine Reedy Miller, MD, and Kia M. Washington University of Pittsburgh, Pittsburgh, PA, USA Background To pioneer vascularized composite allotransplantation (VCA) of the human eye as a clinical reality, our group has developed the first orthotopic model for eye transplantation in the rat for testing of immunomodulation and neuroregeneration therapies. We have also performed human cadaveric studies to design surgical protocols for donor and recipient procedures ahead of advancing to a non-human primate surgical model of eye transplantation Non-invasive methodologies of graft monitoring are paramount to development of this model as well as for clinical practice in future transplantation programs.","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"16 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114357939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}