Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaSupport: NIH, NEI, R44 EY017497, R43 EY021045, and RO1EY06819Program Number: 2061 Poster Board Number: D0200Presentation Time: 11:00 AM - 12:45 PMIn Vivo Administration of Interleukin-2 Increases CornealAllograft Survival through Expansion of CD4+CD25+ TRegulatory CellsMaryam Tahvildari, Parisa Emami-Naeini, Yihe Chen, MasahiroOmoto, Jing Hua, Sunil K. Chauhan, Reza Dana. Schepens EyeResearch Institute, Massachusetts Eye and Ear, Department ofOphthalmology, Harvard Medical School, Boston, MA.Purpose: T regulatory cells (Tregs) have a major role in inhibitinggrowth and differentiation of activated T cells following alloantigenstimulation, and it has been found that interleukin-2 (IL-2) is crucialin the generation and maintenance of these cells. In this study ofmurine corneal transplantation, we aimed to investigate the effect ofsystemic administration of IL-2 on corneal allograft survival.Methods: Corneal transplantation was performed on 8-10 week-oldmale BALB/c (H-2d) mice using C57BL/6 (H-2b) mice as donors.Prior to surgery, recipient mice received 3 intraperitoneal injections(once a day) of either IL-2 (1µg/20 g body weight, in 100µlphosphate buffer saline [PBS]) or PBS alone (as control). Injectionswere continued once daily for one week after surgery followed bytwice a week for another 4 weeks. Allografts were evaluated up to 5weeks post-transplantation and were scored using the standardopacity grading system (from 0 to +5). Frequencies ofCD4+CD25+Foxp3+ Tregs in the draining lymph nodes wereevaluated by flow cytometry on the day of transplantation (day 0),one week (day 7) and two weeks (day 14) afterwards. In addition, thesuppressive function of CD4+CD25+ Tregs derived from draininglymph nodes of recipient mice was evaluated one week aftertransplantation.Results: Of the six corneas transplanted in each group, 66.6% in theIL-2 treated group remained clear up to 5 weeks after surgerycompared to 33.3% in the control group; moreover, IL-2 treatmentdelayed the incidence of graft rejection with mean rejection time of32±1.4 days (mean±SD) for the IL-2 treated vs. 19.5±7.7 days for thecontrol group. Flow cytometric analysis showed significant increasesin the frequencies of CD4+CD25+Foxp3+ Tregs in the draininglymph nodes of recipient mice in the IL-2 treated group compared tothe control group at day 0 (14.54% vs. 10.25%, p=0.014), day 7(14.77% vs. 11.73%, p=0.009) and day 14 (16.15% vs. 12.41%,p=0.0007). Treg suppression assay revealed 16.05% increasedsuppression of effector T cells in the IL-2 treated group compared tothe control group one week after transplantation (p=0.01).Conclusions: According to our results, systemic administration ofIL-2 is capable of enhancing the frequencies and suppressive functionof CD4+CD25+Foxp3+ Tregs, and increases corneal allograftsurvival.Commercial Relationships: Maryam Tahvildari, None; ParisaEmami-Naeini, None; Yihe Chen, None; Masahiro Omoto, None;Jing Hua, None; Sunil K. Chauhan, None; Reza Dana, Allergan(C), Alcon (C), Bausch & Lomb (C), Eleven Bio (I), GSK (F),Novabay (C), Revision Optics (C), Novaliq (C), RIgel (F)Support: NIH Grant EY12963Program Number: 2062 Poster Board Number: D0201Presentation Time: 11:00 AM - 12:45 PMProteomic Analysis of Plasma and Mucosal Samples fromPatients with Stevens-Johnson Syndrome/Toxic EpidermalNecrolysisJulia Malalis 1 , Christine Mata 1 , Daniel Kahn 3 , Amy Lin 1 , Michael J.Mosier 2 , Charles S. Bouchard 1 , Josephine Cunanan 3 , Omer Iqbal 1, 3 ,Debra Hoppensteadt 3 , Jawed Fareed 3 . 1 Ophthalmology, LoyolaUniversity Chicago Stritch School of Medicine, Maywood, IL;2 Surgery, Loyola University Medical Center, Maywood, IL;3 Pathology, Loyola University Chicago Stritch School of Medicine,Maywood, IL.Purpose: Stevens-Johnson syndrome and toxic epidermal necrolysis(SJS/TEN) are life-threatening, immune-complex hypersensitivityreactions that affect the skin and mucous membranes, often resultingin significant ocular inflammation. The purpose of this study was todetermine and compare the proteomic and coagulation profile ofplasma and mucosal discharge samples in affected and unaffectedpatients.Methods: Following institutional review board approval, plasma andswabs from ocular, oral, and skin lesions were obtained from patientswith clinically suspected SJS/TEN (n=5). Two patients had biopsyprovenSJS/TEN. Three patients had alternative skin diagnoses notconsistent with SJS/TEN and were considered abnormal controls.Samples from normal healthy controls were also obtained. Followingcentrifugation, the plasma was frozen at -70°C and later thawed andanalyzed to determine thrombin-antithrombin complex (TAT,Dade®, Marburg, Germany), fibrinopeptide (F1.2, Dade®),plasminogen activator inhibitor-1 (PAI-1, Diagnostica Stago®,Asnieres Sur Seine, France), ZYMUPHEN platelet microparticleactivity (Hyphen® BioMed, Neuville-Sur Oise, France),HEMOCLOT protein C (Stago®), and STACHROME antithrombin(Stago®), using ELISA kits as per manufacturer’s instructions. Theswabs were immediately frozen at -70°C and later thawed. Thedischarges were isolated following addition of 0.25 ml of saline toeach swab and double centrifugation. The discharges and plasmasamples were analyzed using SELDI-TOF technique.Results: Analyses of the SJS/TEN plasma samples revealed amarked increase in the TAT complexes (6.3±5.9µg/ml), F1.2(430.4±202.4 pmol/L), platelet microparticles (13.1±9.3nM) andprotein C levels (90.5±63.4%), with a corresponding decrease in PAI-1 (53.3±18.8ng/ml) and antithrombin levels (80.7±42.4%) comparedto normal control human plasma, suggesting a procoagulant state.Protein chip array of the SJS/TEN skin and oral mucosal samplesexhibited two major peaks at 14.2 kDa and 15.6 kDa, in the samemolecular weight range as recombinant human granulysin, amolecule implicated in the pathophysiology of SJS/TEN. Thesepeaks were not present in the control group.Conclusions: Procoagulant factors and unique peaks suggestive ofgranulysin may lead to the development of targeted therapy aimed toattenuate local and systemic inflammatory processes in patients withSJS/TEN.Commercial Relationships: Julia Malalis, None; Christine Mata,None; Daniel Kahn, None; Amy Lin, None; Michael J. Mosier,None; Charles S. Bouchard, None; Josephine Cunanan, None;Omer Iqbal, None; Debra Hoppensteadt, None; Jawed Fareed,NoneSupport: Illinois Society for the Prevention of Blindness (ISPB)GrantProgram Number: 2063 Poster Board Number: D0202Presentation Time: 11:00 AM - 12:45 PMMorphologic Dendritic Immune Cells Parameters RevealDifferential Characteristics between the Central and PeripheralCornea: an In Vivo Confocal Microscopy Normative DataClara M. Colon 1 , Bernardo M. Cavalcanti 1, 2 , Shruti Aggarwal 1 ,Andrea Cruzat 1, 2 , Candice Williams 1 , Douglas Critser 4 , Amy Watts 3 ,Christine W. Sindt 4 , Pedram Hamrah 1, 2 . 1 Department of©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.