Scottish Society of Cytomics SSC 2014
Scottish Society of Cytomics SSC 2014
“Transla onal Cytometry from Bench to Bedside”
1st Annual Mee ng on 25th September 2014
With an exci ng line up of Speakers, Workshops & Awards!
Ins tute of Medical Sciences Level 7 Conference Room University of Aberdeen Foresterhill, Aberdeen
Local Organisers: Linda Duncan, Raif Yuecel, David Wilson & Janice Forsyth
Free Registra on and Lunch kindly provided by generous sponsors: Please send your registra on to [email protected] “Transla onal Cytometry from Bench to Bedside”
KEY SPEAKERS TOPICS Paul Smith (University of Cardiff) Regenera ve Medicine Derek Davies (CRUK London) Cancer Research Ian Dimmick (University of Newcastle) Drug Discovery Cosimo De Bari (University of Aberdeen) Clinical Cytometry John Campbell (NSL Edinburgh) Immunology Adriano Rossi (MRC Edinburgh) Data Analysis Tools Elizabeth Ballou (University of Aberdeen) Mul colour Cytometry David Wilson (NHS Grampian) Fungal Research
SPECIAL LECTURE: Paul Smith Past President of ISAC & Emeritus Professor of Cancer Biology University of Cardiff Cytomics Of The Tumour Microenvironment: Therapeu c Targe ng or “Loca on, Loca on, Loca on!”
SPECIAL WORKSHOP: Ian Dimmick Manager, Flow Cytometry Core Facility, Newcastle upon Tyne University Prac cal session on 8-10 colour Compensa on
1st Annual meeting of SSC 2014 Contents
6 SSC Programme for 25th of September
9 Speaker Biographies
16 Sponsors and Exhibi on Plan
20 Poster Presenta ons & Abstracts
27 SSC Membership Form
28 Direc ons and Map of IMS
29 Aberdeen City Map
31 SSC Conference Commi ee
32 Post-SSC Mee ng Workshop on 26th of September “Flow Cytometry Data Analysis Workshop”
1st Annual meeting of SSC 2014 “Ceud Mìle Fàilte gu Aberdeen” - "A Hundred Thousand Welcomes to Aberdeen”
Dear Par cipant, Dear Colleagues,
On behalf of the Sco sh Society of Cytomics (SSC), it is my greatest pleasure to welcome you to our 1st Annual mee ng of the newly formed Society SSC in Aberdeen. Following the Scotland and Northern England Flow Cytometry mee ng 2012 in Moredun, we considered the establishment of a new cytometry group in Scotland, that brings scien sts in the field of Cytomics together from the northern regions.
Why Cytomics? Actually the theme of our first SSC mee ng already provides the answer —Transla onal Cytometry from Bench to Bedside— relates to the future of this society and it reflects our shared passion for cellular systems. Cytometry is growing as a diagnos c and monitoring technology in the research and clinical laboratory and that it will be useful to establish a forum to exchange informa on, to present research data and to explore new applica ons of the rapidly expanding technologies of image and flow cytometry. To be er understand and further explore the cellular systems, the merging and bundling of technologies is necessary. Single technology can’t provide the ul mate solu on! Therefore CYTOMICS, which analyzes the func onal rela onship between cell systems and networks using the “omics” technologies (e.g. Genomics, Proteomics), will give a comprehensive view of a disease. The analysis of already exis ng data from scien fic studies or rou ne diagnos c procedures makes cytomics a valuable tool for studying diseases, that will be of immediate value in transla onal medicine.
The Sco sh Society of Cytomics was formed with the inten on of providing informa on related to the art of cytometry (flow and imaging) including its cross technology applica ons to genomics, proteomics, in vivo imaging and other core related areas between the research organiza ons in Scotland and Northern England. We believe that SSC will be the ideal forum for users of cytometry & omics systems to get together, share exper se and innova on, discuss pi alls, successes and ideas for the future of transla onal cytomics.
Simply fostering the strong network of this scien fic community!
1st Annual meeting of SSC 2014
It is a great privilege for me to announce this exci ng line up of talks, exhibi ons and workshop. SSC 2014 brings together top scien sts from various disciplines, and interna onal renowned keynote speakers. On behalf of the organizing commi ee, I would like to express my sincere gra tude to our excep onal speakers for taking the me to a end and support our first SSC mee ng.
Further to our talks we will have poster session during the coffee and lunch break. This poster presenta on will results in poster awards, from generous sponsors, to recognize the excellent research done by scien sts.
Therefore, we encourage you to visit the stands of the commercial exhibi on as without their support this mee ng wouldn’t be such a success. Please take this this opportunity to acknowledge the sponsors for their support and helping us. Our special thanks goes to Becton Dickinson, Sony, BioStatus and Miltenyi Biotec for their extra dona ons and contribu ons.
This mee ng is not the outcome of one person; it’s the result of a team. I would like to express my gra tude to the fantas c organizing commi ee and members, namely Lynne Lumsden, Janice Forsyth, Ian Dimmick, Mara Rochi, and David Wilson. My big huge thank you to Linda Duncan, who has done the majority of the administra ve and exhibi on organiza on, nothing would be possible without her.
Finally we THANK YOU for par cipa ng in this first Sco sh Society of Cytomics mee ng. We hope that this conference will bring about rich and rewarding experiences for all of us in science, and help develop friendships.
Let us greet old friends and make new ones! Once again, I welcome you to Aberdeen, and have a good me at SSC 2014.
Raif Yuecel, IFCC University of Aberdeen Chairman SSC
"Coming together is a beginning; keeping together is progress; working together is success" -Henry Ford
1st Annual meeting of SSC 2014 Programme Thursday 25th September
08:15 Coffee Recep on and Registra on (IMS level 7 CR)
09:00 Welcome address by Raif Yuecel, SSC, IFCC, University of Aberdeen Peter McCaffery, IMS Director, University of Aberdeen Alistair Brown, Integrated Centre, University of Aberdeen
Speakers
09:30 Derek Davies, CRUK London Sharing the love: Networking amongst cytometrists
09:45 Elizabeth Ballou, University of Aberdeen FACS for Fungi: Revealing popula on heterogeneity and dynamics in fungal pathogens via flow cytometry
10:00 Ian Dimmick, Newcastle upon Tyne University A unique approach to compensa on, and its verifica on
10:20 David Wilson, NHS Grampian Quality in Clinical Cytometry - do we con nue to make the same mistakes?
10:40 COFFEE & Poster Session in IMS Atrium
11:00 Derek Davies, CRUK London Drug effects in mul ple rounds of cell division monitored by flow cytometry
11:20 Cosimo De Bari, University of Aberdeen Cell therapies for joint repair
11:40 John Campbell, Na onal Science Laboratory Edinburgh Transla ng Research Flow Assays for Characteriza on of Cellular Therapeu cs
12:00 Adriano Rossi, MRC Edinburgh Flow cytometric techniques for isola ng and analysing leucocytes
1st Annual meeting of SSC 2014 6 12:20 Special Lecture: Paul Smith, University of Cardiff Cytomics Of The Tumour Microenvironment: Therapeu c Targe ng, or Loca on, Loca on, Loca on!
13:00 LUNCH & Exhibi on & Poster Viewing (IMS Atrium)
AFTERNOON Parallel Sessions
14:00 – 15:00 GROUP I:Prac cal session on 8-10 colour Compensa on by Ian Dimmick , IMS, IFCC lab 2.54
15:00 – 16:00 GROUP II:Prac cal session on 8-10 colour Compensa on by Ian Dimmick , IMS, IFCC lab 2.54
14:00 - 16:00 New technologies by industrial applica on specialists at IMS level 7 CR: Please see following page
16:00 - 17:00 SSC Commi ee mee ng (open mee ng, IMS level 7)
17: 30- Come together over Wine and Cheese & Poster Award Winner Announcement (IMS level 7, everyone welcome)
1st Annual meeting of SSC 2014 7 A ernoon parallel session from 14:00 – 16:00 at IMS level 7 CR New technologies by industrial applica on specialists (10min presenta ons!)
14:00 Roy Bongaerts, Sony Biotechnology Europe New mul colour single cell technology: Spectral cell analysis on the Sony SP6800
14:11 Laura Schneider, Cell Signaling Technology, NEB Ltd Considera ons to study cell signaling by flow cytometry
14:22 Stephen Rackstraw, BD Biosciences Sor ng the Molecules that Count’ - Single Cell Sor ng for Next Genera on Sequencing’
14:33 Kalpana Singh, Miltenyi Biotech Ltd MACS Flow Cytometry Products: Innovate Solu ons for all Your Flow Needs
14:44 Lindsey Ward, iLab Solu ons LLC Addressing the Challenges of Opera ng a Core Facility
14:55 Leonor Heleno Wielgosz, Stratocore Facility Management bringing the focus back to Research
15:06 Jasmin Moss Life Technologies, Thermo Fisher A une® NxT: The next genera on in acous c focusing cytometry
15:17 Sco Cribbes, Nexcelcom Ltd. Adherent Imaging Cytometry Complemen ng Flow Cytometric Measurements to address the diverse needs of a research environment
15:28 Andrea Valle, De Novo So ware Cytometry Data Analysis using FCS Express
15:39 Norman Maidment, Beckman Coulter Ltd Going Dry: Alterna ve monoclonal an body op ons for Flow Cytometry
15:50 Alistair Stewart, Affymetrix – eBioscience TBC
8 1st Annual meeting of SSC 2014 Speakers
Paul%Smith% Emeritus)Professor)of)Cancer)Biology)) Institute)of)Cancer)&)Genetics,)Innovation)and)Engagement) Representative,)) Cardiff)University,)UK) ) Paul)has)been)active)in)academic)research)in) the) fields) of) DNA) repair,) drug) development,) cytometry,)biochip)and)imaging)for)more)than) 30)years)at)the)MRC)Centre)in)Cambridge)and) at)Cardiff)University)where)he)is)now)Emeritus) Professor) of) Cancer) Biology.) His) research) expertise) has) focused) on) cancer) K) encompassing)the)cell)cycle,)cancer)stem)cells) and) resistance) to) anticancer) drugs.) His) inventions) include) a) range) of) anticancer) drugs) and) molecular) probes) including) the) DRAQ) dyes.) He) has) served) the) International) Society) for) Advancement) of) Cytometry,) being) the) President) from) 2010K12) and) successfully) introducing) the) CYTO) series) of) congresses) with) linked) educational) initiatives.) Earlier,) he) was) responsible) for) the) ISAC’s) 1st) International)Cytomics)Meeting)in)Wales)in)2003,)aimed)at)introducing) the) concept) of) cytomics.) His) entrepreneurial) activities) include) the) coK founding)of)the)technology)companies)Biostatus)Ltd)and)Biosuspensions) Ltd.) He) is) the) Science) Director) of) the) awardKwinning) startKup) Oncotherics) Ltd) –) a) new) pharmaceutical) company) that) has) exploited) cytometric) techniques) in) the) development) of) novel) hypoxiaKactivated) proKdrugs)for)the)treatment)of)a)wide)range)of)cancers.) )
1st Annual meeting of SSC 2014 9 Derek%Davies% Manager'of'the'FACS'Laboratory,'London'Research'Institute'(LRI),'' Cancer'Research'UK,'London,'UK' ' ' Derek’s' first' post' after' graduating' was' in' a' regional'Cytopathology'Unit'where,'amongst' other'duties,'he'was'introduced'to'cytometry'' F'using'a'microdensitometer'to'measure'DNA' content' in' cervical' cells.' This' led' to' his' first' exposure' of' a' flow' cytometer' –' the' FACS' Analyzer.' Realizing' the' potential' for' cytometry' in' cancer'diagnosis'and'research,'he'moved'to' Kings'College'Hospital'in'London'on'an'MRCFfunded'project'to'look'at' the'feasibility'of'using'a'flowFbased'preFscreen'in'cytological'specimens.' In' 1990,' he' moved' to' the' Imperial' Cancer' Research' Fund' to' work' in' what' was' then' a' small' core' facility' providing' support' for' 40' research' Laboratories.'Derek'became'Head'of'that'unit'in'1996'and'remains'there' today.'Derek'currently'manages'a'team'of'7'staff'and'9'cytometers'(6' analysers'and'3'sorters).' Derek'is'Chair'of'flowcytometryUK,'he'sits'on'the'Council'of'the'Royal' Microscopical'Society'and'head'of'the'ISAC'Core'Managers'TaskForce.' He'is'very'active'in'teaching'cytometry'within'the'UK'and'coForganises' an'annual'meeting'on'behalf'of'the'RMS'as'well'as'speaking'at'several' other'national'and'international'courses.' Current'Research'Interest:' Development'of'instrumentation'and'methodologies'appropriate'to'the' needs'of'a'cancer'research'institute.' ' '
1st Annual meeting of SSC 2014 10 Elizabeth*Ballou** Research( Fellow( in( the( Aberdeen( Fungal( Group( at( Institute( of( Medical( Sciences,(University(of(Aberdeen,(UK( ( Elizabeth( received( her( PhD( in( Genetics( and( Genomics( from( Duke( University,( USA,( in( the( lab( of( Dr.( Andrew( Alspaugh,( where( she( studied(the(role(of(conserved(RhoGGTPases(in( C.(neoformans(thermotolerance,(polarity,(and( ploidy.( As( a( member( of( Prof( Alistair( Brown's( lab( in( the( AFG,( Elizabeth( has( used( flow( cytometry( and( FACS( analysis( to( study( the( impact( of( host( carbon( source( on( C.( albicans( morphogenesis(and(pathogenicity.(( Current(Research(Interest:( Her(work(focuses(on(the(impact(of(the(human(host(environment(on(the( fungal(pathogens(Candida(albicans(and(Cryptococcus(neoformans.(
David&Wilson&& Principal)Healthcare)Scientist,)Immunology)Laboratory,)Aberdeen)Royal) Infirmary,)UK) He)has)over)thirty)years)experience)in)clinical) flow)cytometry) working) both) in) Aberdeen) and)Glasgow.))David)is)a)founder)member)of) the) British) Society) for)Histocompatibility) and)Immunogenetics) and) is) a) Fellow) of) the) Institute) of) Biomedical) Science) and) a) member) of) their) Immunology) Scientific) Advisory) Panel.)) His) interests) in) addition) to) flow)cytometry) include) transplantation) and) autoimmune)diseases.)
1st Annual meeting of SSC 2014 11 Ian$Dimmick$ Manager,(Flow(Cytometry(Core(Facility,(Newcastle(upon(Tyne(University,( UK( ( (Ian( began( his( career( in( a( clinical( setting( and( from(1972(to(1991(worked(in(various(research( posts(using(flow(cytometry(as(a(diagnostic(and( research( tool( primarily( for( HIV,( leukaemia,( lymphoma( and( a( broad( range( of( immunological(testing.( ( From( 1991( to( 2006( Ian( worked( as( a( Flow( Cytometry( Applications( and( Instrumentation( Specialist(for(various(commercial(companies.( Ian( took( up( his( current( post( in( 2006( and( manages( the( flow( cytometry( core( facility( in( Newcastle( upon( Tyne( University.( In( his( current( role( Ian( organises( various( flow( cytometry( teaching( courses( and( conferences,( such( as( the( quarterly( ‘Colour( Compensation’( course( and( the( annual( ‘Practicalities(of(Flow(Cytometry/Cellular(Analysis’(meeting,(both(held(in( Newcastle(upon(Tyne.( Current(Research(Interests( Research(and(clinical(flow(cytometry(in(particular(leukaemia,(lymphoma( and(stem(cells.(
1st Annual meeting of SSC 2014 12 Cosimo&De&Bari& Professor'of'Translational'Medicine'&'Hon'Consultant'Rheumatologist' Institute'of'Medical'Sciences,'University'of'Aberdeen,'UK' ' Cosimo' is' a' clinically' active' rheumatologist' with' expertise' in' regenerative' medicine' for' musculoskeletal'applications.' Cosimo'graduated'in'Medicine'(maxima'cum' laude)' from' the' University' of' Bari' (Italy),' where' he' also' underwent' specialist' training' in' Rheumatology.' ' He' then' moved' to' Belgium,'where'he'obtained'his'PhD'from'the' University'of'Leuven'and'was'recipient'of'the' Rotary' Young' Investigator' Award' 2003' from' the' Belgian' Society' for' Rheumatology.''His'work'in'Belgium'contributed'to'the'development'of' an' autologous' cellular' product' for' articular' cartilage' repair,' which' recently' became' the' first' cellPbased' product' to' obtain' EU' Marketing' Authorisation'as'an'Advanced'Therapy'Medicinal'Product.' In'2003'Cosimo'moved'to'the'UK'in'the'Department'of'Rheumatology'at' King's'College'London.''In'May'2005'he'was'awarded'a'Clinician'Scientist' Fellowship'from'the'Medical'Research'Council'and'in'December'2005'he' was'appointed'Clinical'Senior'Lecturer'&'Consultant'Rheumatologist.' Since'September'2007'Cosimo'is'Professor'of'Translational'Medicine'at' the' University' of' Aberdeen,' where' he' currently' leads' the' Musculoskeletal'Research'Programme.' Cosimo' has' expertise' in' stem' cell' research' for' musculoskeletal' repair,' regenerative' medicine' and' tissue' engineering.' ' Current' research' interests' in' his' Group' include' (i)' the' development' of' stem' cellPbased' tissue'engineering'products'for'cartilage'and'bone'repair;'(ii)'the'study' of' the' stem' cell' niches' in' the' joint' in' health' and' diseases' such' as' osteoarthritis' and' rheumatoid' arthritis;' (iii)' the' investigation' of' the' developmental'ontogeny'of'mesenchymal'stem'cells.'
1st Annual meeting of SSC 2014 13 Adriano(G(Rossi( Professor'of'Respiratory'and'Inflammation'' MRC' Centre' for' Inflammation' Research,' Queen's' Medical' Research' Institute,'University'of'Edinburgh,'UK' ' Adriano' G.' Rossi' received' his' BSc' and' PhD' from'the'University'of'Glasgow,'Scotland'UK.'' He'carried'out'postdoctoral'research'at'Wake' Forest'University,'North'Carolina,'USA'and'at' the'National'Heart'&'Lung'Institute,'London,' UK.'' He'currently'holds'a'Chair'in'Respiratory'and' Inflammation'Pharmacology'at'the'University' of' Edinburgh,' with' a' research' focus' on' the' mechanisms'regulating'the'resolution'of'inflammation.'He'was'awarded' a'DSc'from'the'University'of'Edinburgh'and'is'a'Fellow'of'the'Society'of' Biology'and'a'Fellow'of'the'British'Pharmacological'Society.'' Research'Theme:'Immune'Modulation'and'Regulation'of'Inflammation,' Imaging' Inflammation.' We' aim' to' gain' a' better' understanding' of' the' mechanisms' controlling' inflammatory' processes' with' a' view' to' help' develop'novel'therapies'for'chronic'inflammatory'diseases.''For'this'we' aim' to' elucidate' the' mechanism' regulating' inflammatory' cell' behavior' and'apoptosis'and'manipulate'the'processes'controlling'the'resolution' of' inflammation' in' order' to' develop' new' therapeutic' strategies' to' remove'unwanted'and'dysregulated'inflammation.' '
1st Annual meeting of SSC 2014 14 John%Campbell% Associate) Director) at) Research,) Development) &) Innovation,) SNBTS,) National)Science)Laboratory,)Hon)Reader)at)University)of)Edinburgh,)UK) ) John) is) Associate) Director) of) Research) and) Development) at) the) Scottish) National) Blood) Transfusion)service)(SNBTS))in)Edinburgh.)He) completed)his)PhD)in)Pathology)at)Edinburgh) in) 1995) on) the) immunopathogenesis) of) lymproliferative) disease,) and) has) worked) in) the)cellular)therapy)field)for)over)20)years.)) He) has) held) various) academic) positions) including)Lecturer)in)Tumour)Immunology)at) the)University)of)Glasgow.)He)has)also)worked)extensively)in)the)biotech) industry)as)R&D)project)leader,)Group)Manager)Clinical)Science)and)Cell) Analysis,)and)Director)of)Clinical)Immunology)at)Miltenyi)Biotec)GmbH.) He)returned)to)full)time)academic/healthcare)work)at)the)end)of)2012.)) John) is) currently) the) national) head) of) research) for) SNBTS) and) is) Hon.) Reader)at)the)University)of)Edinburgh.)SNBTS)has)a)substantial)cellular) therapy)research)programme,)with)over)25)full)time)scientists)working) on)basic)cellular)function;)translation)of)laboratory)protocols)to)full)GMP) processes;) and) production) of) cellular) therapeutics) for) treatment) of) patients.) SNBTS) has) a) dedicated,) fully) MHRA) licensed,) GMP) cellular) therapy) production) centre) at) the) Scottish) Centre) for) Regenerative) medicine.) Cellular) therapeutics) currently) in) early) phase) trials) include) haematopoietic)stem)cells)for)liver)repair,)Corneal)Limbal)Stem)Cells)and) EBVYspecific)Cytotoxic)T)Lymphocytes)for)PTLD.)) John’s) own) research) programme) focuses) on) the) generation) of) novel) methodologies) to) characterise) and) improve) the) efficacy) and) safety) of) novel)cellular)therapeutics)in)vivo,)including)targeting)of)injected)cells)to) specific)anatomical)sites.) )
1st Annual meeting of SSC 2014 15 1st SSC Mee ng Venue We are grateful to the Ins tute of Medical Sciences at the University of Aberdeen for their support and providing us an excellent loca on for our first SSC conference.
1st Annual meeting of SSC 2014 16 Sponsorlist We would like to acknowledge the following sponsors for their generous support and helping us to make this conference a big success:
1st Annual meeting of SSC 2014 17 Extra Dona ons We are especially grateful to the following contributors for their extra dona ons and support to SSC 2014.
Dona on of An bodies
Dona on for Catering
Dona on of 1st Poster Prize
Dona on of 3rd Poster Prize
1st Annual meeting of SSC 2014 18 Exhibi on Plan in IMS Atrium The IMS Atrium is open the en re day. Lunch and Poster Presenta on will take place in the exhibi on area.
1. Beckman Coulter
2. Affymetrix – eBioscience
POSTER Area 3. Sysmex-Partec 1 19 4. Stratocore
2 5. Sony 18 3 6. New England BioLabs LUNCH COFFEE 7. Merck Millipore 17 4 8. iLab Solu ons 5 16 9. Life Technologies Table LUNCH 10. Caltag Medsystems
6 11. BioRad
12. Cambridge Bioscience 7 13. Enzo Life Sciences 8
Coffee Shop 14. Biotechne
9 15. BioLegend
10 16. Becton Dickinson
17. Nexelcom 11 18. DeNovo So ware 15 14 13 12 19. Miltenyi Biotec
1st Annual meeting of SSC 2014 19 Poster Presenta ons
The posters will be presented during the coffee &lunch breaks in the IMS atrium and the winners will be announced during the closing ceremony at 5:30pm at IMS level 7. We will recognize the excellent research by scien sts who have applied cytometry (flow and/or imaging) in their studies by offering poster awards. Cash prizes and a cer ficate will be awarded to the winners: •1 st Poster prize £400 •2 nd Poster prize £200 •3 rd Poster prize £100 All scien sts who are students or postdoctoral researchers (who have received their doctorate within the last five years) were invited to submit their poster.
POSTER 1 Dele on of myeloid-PTP1B decreases MHC Class I expression and pep de presenta on through an IL-10 dependent mechanism in response to LPS challenge.
Samantha Le Sommer, Cris na Mar n-Granados, Mirela Delibegovic Ins tute of Medical Sciences (IMS) School of Medical Sciences, University of Aberdeen, UK
Protein Tyrosine Phosphatase 1B (PTP1B) inhibi on is a target in the treatment of type 2 Diabetes Mellitus, and as such PTP1B inhibitors are in Phase II clinical trials. Previously our lab demonstrated that myeloid-specific dele on of PTP1B (LysM PTP1B) results in an increase in systemic IL-10 secre on and expression. In the current work we inves gated how PTP1B deficiency affects the ac va on phenotype of murine macrophages in response to inflammatory s muli. We demonstrate that myeloid-specific PTP1B dele on results in a decrease in expression of MHC Class I, along with co-s mulatory molecules CD80 and CD40. Interac on assays reveal a defect in the cells’ ability to ac vate reporter B3Z T cells. Myeloid-specific PTP1B dele on increases the percentage of bone-marrow-derived-macrophages (BMDMs) posi ve for IL-10 which is associated with a decrease in iNOS produc on. Western blo ng analysis demonstrated hyper-phosporyla on of ERK1/2 which has been suggested before to improve access to the IL-10 promoter. This provides evidence to suggest that myeloid-PTP1B dele on decreases MHC Class I expression and pep de presenta on through an IL-10- dependent mechanism.
1st Annual meeting of SSC 2014 20 POSTER 2
CCL19-sorted mature dendri c cells have enhanced lymph node migratory capacity and func on.
Michelle L. Le Brocq (1), Alasdair R. Fraser (1,2), Gerard J. Graham (1) and John D.M. Campbell (1,2) (1) Chemokine Research Group, Ins tute of Infec on, Immunology and Inflamma on, University of Glasgow, Glasgow, G12 8TA, UK. (2) Sco sh Na onal Blood Transfusion Service, Cellular Therapeu cs Research Group, Edinburgh, EH17 7QT, UK.
Cell therapy regimens are frequently compromised by low-efficiency cell homing to therapeu c niches. Improvements in this regard would enhance effec veness of clinically-applicable cell therapy. The major regulators of ssue-specific cellular migra on are chemokines and therefore selec on of therapeu c cellular popula ons for appropriate chemokine receptor expression would enhance ssue-homing competence. We have recently shown that bio nylated ligands for chemokine receptors can be used as efficient cell sor ng reagents (J.Immunol 192:6120 (2014)). These ligands have several advantages over an bodies, as they can be chemically synthesized to the appropriate grade for clinical use; produced and modified extremely quickly and are generally not species-specific. Furthermore, receptors are rapidly recycled post cell selec on leaving cells free to respond to chemokine gradients, and thereby migrate to the required therapeu c site. We have applied this technology to the improvement of dendri c cell (DC) and T cell products for poten al use as cellular therapeu cs. CCL19, a high-affinity ligand for CCR7, is of par cular interest in this field, as CCR7 expression by DC is a marker of maturity and lymph node homing ability. Also, CCR7+ central memory T cells are the most desired cell popula on for genera on or engineering of tumour-an gen-specific therapeu c responses. Here we report the genera on of bio nylated, chemically synthesised, modified human CCL19 molecules for op mised cell sor ng. Our results demonstrate that these ligands allow the efficient enrichment of mature, an gen-loaded human or murine CCR7+ DC using a rapid and established magne c bead sor ng strategy. CCR7+ murine bone marrow-derived DC (BM-DC) isolated using this GMP-compliant method display significantly improved lymph-node homing ability in vivo over standard (unsorted) BM-DC prepara ons.
1st Annual meeting of SSC 2014 21 Using in vitro models, such as murine OT- I, we also demonstrate the ability of these sorted DC to efficiently generate both effector and memory an gen- specific T cell responses. In conclusion, we show that bio nylated ligands for chemokine receptors cons tute a novel, GMP-compliant cell sor ng strategy that could enhance the efficacy of exis ng clinical cell therapy regimens.
POSTER 3
Ac va on of immune responses using ovine len virus vaccine vectors.
Rebecca McLean1,2, Ann Wood1, Jayne Hope2, Gary Entrican1, David Griffiths1 1) Moredun Research Ins tute, Edinburgh 2) The Roslin Ins tute and Royal (Dick) School of Veterinary Studies, University of Edinburgh
Many infec ous diseases of livestock are s ll not preventable due to the lack of appropriate vaccines and it is widely recognised that new approaches to vaccine design are required to address this need. The aim of this project is to develop ovine len viruses as vaccine vectors for use in ruminants. In recent years, a number of viral vectors have been developed for use as vaccine delivery systems. Len viruses are a rac ve candidates as vaccine vectors due to their capacity to efficiently transduce non-dividing cells and to maintain stable, long-term expression of transgenes. In addi on, len viral vectors have been shown to be able to produce an an gen mediated immune response in mice, suppor ng their con nued development as vaccine vectors. To date, the use of len viral vaccine vectors has not yet been inves gated in livestock. This project will focus on the development of a safe, efficient len viral vector for ruminants with par cular interest in understanding the mechanisms of the immune responses. Experiments performed so far have led to the produc on of novel self-inac va ng and integra on deficient vaccine vectors.
1st Annual meeting of SSC 2014 22 POSTER 4
Characterisa on of the murine C-type lec n receptor CLECSF8 (MCL) and its role in an -mycobacterial immunity
Kerscher B.1, Wilson G. 1, Reid D. 1, Yamasaki S. 2, Willment J. 1, Brown G. 1 1University of Aberdeen, Ins tute of Medical Sciences, Aberdeen Fungal Group, Aberdeen 2Kyushu University, Research Center for Infec ous Diseases, Fukuoka
Introduc on C-type lec n-like receptors (CTLRs) play cri cal roles in immunity and homeostasis by recognising a great variety of microbial or endogenous ligands. CLECSF8 is a member of the Dec n-2 family of C-type lec ns. Previous research indicates that CLECSF8 associates with the signalling adaptor FcRγ which is essen al for signalling through the SYK and CARD9 pathway. Recently, the mycobacterial cord factor (TDM, trehalose-6,6’- dimycolate) was iden fied as the ligand of CLECSF8, as shown previously for the closely related CTLR Mincle. Objec ves The aim of this study was to characterise the expression of the murine CLECSF8 receptor in different ssues and cell types under steady state, s mulated or infec on condi ons rela ve to Mincle and Dec n-2. Materials and Methods We generated and characterised rat monoclonal an bodies (mAb) against mCLECSF8. Using mCLECSF8 mRNA expression data (rt-PCR) as a guideline, mCLECSF8 protein expression was inves gated by flow cytometry. Results Ini al characterisa on of the an -mCLECSF8 an bodies confirmed their func onality in flow cytometry and immunoblo ng experiments. Using a mCLECSF8 reporter cell line, we found one mAb clone (3A4) that blocks receptor mediated Mycobacterium bovis BCG recogni on without ac va ng the receptor which will providing a useful tool for in vivo blocking experiments. While mCLECSF8 transcript was found to be widely expressed, CLECSF8 protein expression was found predominantly on neutrophils and/or macrophages within the peritoneal cavity, blood and bone marrow. Interes ngly, basal CLECSF8 expression in the lung was very low, but was strongly upregulated during BCG infec on.
1st Annual meeting of SSC 2014 23 In vitro, CLECSF8 expression was strongly induced by TLR antagonists or microbial s muli and expression profiles between CLECSF8 and Mincle were highly similar. In agreement with previous reports our data suggests that mCLECSF8 associates with the FcRγ chain. Co-expression experiments in a murine fibroblast cell line indicate Mincle, but not Dec n-2, forms a heterodimer with CLECSF8 similar to their rat homologues. Conclusion Murine CLECSF8 was predominantly expressed on neutrophils and macrophages in a variety of ssues and can be induced by microbial s muli.
POSTER 5 Development of a FACS-based Smac/DIABLO release assay to monitor individual drug responses and to pave the way towards pa ent-tailored therapies
Jigyasa Arora, Ralf Michael Zwacka, Andrea Mohr School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, United Kingdom
The induc on of programmed cell death (apoptosis) in tumour cells is an essen al cellular and molecular process in response to cancer treatment, and important for its success. Resistance to such therapies can occur at different molecular levels. One essen al step in apoptosis signalling is the release of several factors from mitochondria into the cytosol. In this context, Smac/DIABLO, aside from cytochrome c, has been recognised as one of these key pro-apopto c factors. Smac/DIABLO binds to the an -apopto c factor XIAP in the cytosol thereby blocking its capacity to inhibit ac va on of caspases. As XIAP is known to be up-regulated in many cancer cells leading to treatment resistance, it is important to be able to monitor drug responses on an individual and single cell level in primary tumour material. To this end, we developed a FACS-based Smac/DIABLO release assay. Using the cytotoxic drug 5-Fluorouracil, we could show that the signal for Smac/DIABLO increased, reflec ng the appearance of Smac/DIABLO in the cytosol. This signal shi could be inhibited by addi on of the pan-caspase inhibitor zVAD. Moreover, these results correspond to data from biochemical analysis of mitochondrial and cytosolic frac ons that also show a release of Smac/ DIABLO into the cytosol. Thus, this FACS method represents a generally useful tool to understand apoptosis signalling and resistance in cancer cells, which can inform treatment op ons for individual pa ents.
1st Annual meeting of SSC 2014 24 POSTER 6
TGF-β2 mediated secre on of sCTLA-4 from regulatory T cells
Khanolkar R1, Rajpara S2, Muller F2, Depasquale I3, Lawson L2, Barker RN1, Nicolson M4, Ormerod AD1,2 and Ward FJ1 1Division of Applied Medicine, Ins tute of Medical Sciences, Foresterhill, AberdeenAB25 2ZD UK. 2Department of Dermatology, Aberdeen Royal Infirmary, Aberdeen AB25 2ZR UK. 3Department of Plas c surgery, Aberdeen Royal Infirmary, Foresterhill, Aberdeen AB25 2ZN UK. 4Department of Oncology, Aberdeen Royal Infirmary, Foresterhill, Aberdeen AB25 2ZN UK.
Cytotoxic T lymphocyte an gen-4 (CTLA-4), a membrane bound inhibitory receptor whose expression is induced on ac vated T cells, has been established as an important regulator of T cell responses and serves to maintain peripheral tolerance. Addi onally, recent characteriza on of a novel mechanism of extrinsic suppression mediated by the soluble isoform of CTLA-4 (sCTLA-4) has served to further establish the importance of CTLA-4 in maintaining homeostasis of the immune system. Previously, we have shown that selec ve blockade of sCTLA-4 in pa ents with metasta c melanoma enhances immune responses including increased an gen-specific prolifera on of both CD4+ and CD8+ T cells, compared with an isotype an body control or pan CTLA-4 blockade. Selec ve blockade also enhanced T cell effector cytokine responses, notably IFN-γ and IL-17. In this study, we demonstrate a protocol for genera ng a regulatory T cell popula on, which is characterised by the produc on of high levels of sCTLA-4. Treatment of naïve human T cells with TGF-β2 together with an an -CD3 mAb/IL-2 T cell s mulus increased their capacity to produce high levels of sCTLA-4, while decreasing produc on of effector cytokines - IFN-γ, IL-17 and IL-10. Furthermore, analysis of these sCTLA-4 producing T cells has shown that they express the T regulatory cell transcrip on factor – FoxP3. Taken together, our data show that TGF-β2 could serve as an a rac ve therapeu c tool to alleviate symptoms of autoimmunity through the induc on of regulatory T cell popula ons that secrete immunosuppressive sCTLA-4; while on the other hand, neutralizing the effects of TGF-β2 could also prove beneficial to pa ents with cancer.
1st Annual meeting of SSC 2014 25 POSTER 7
Can Selenium supplementa on modify oxida ve stress in-vitro? A role for selenium supplementa on in the preven on of Cardiovascular Disease
Dean Leighton, Marie Goua, Eimear Dolan, Katherine Burgess, Giovanna Bermano* Ins tute of Health and Wellbeing Research, Robert Gordon University, Aberdeen *Corresponding author
Two thirds of the UK popula on are either overweight (body mass index (BMI) 25-29.9 kg/m2) or obese (BMI >30 kg/m2). Such individuals are characterised by systemic oxida ve stress (OS), which is deemed to play a key role in cardiovascular disease (CVD). OS results from reduced an oxidant status and increased reac ve oxida ve species (ROS) forma on. An increased dietary an oxidant intake or up-regula on of endogenous an oxidant enzymes may counteract this OS state and therefore lower CVD risk. Selenium is an essen al dietary micronutrient incorporated within the cataly c site of the endogenous an oxidant Glutathione Peroxidase (GPx) enzymes, which protect cells from OS and consequent cell damage. There is, however, a lack of knowledge concerning the effect of selenium supplementa on in an OS state representa ve of sedentary overweight/ obese individuals. The aim of this work was to inves gate the impact of selenium supplementa on to modify monocyte cell viability/ROS produc on under OS. U937 monocyte cells were supplemented with sodium selenite (100nM or 200nM) or not and cultured for 48 hours at 37°C. Paraquat (100mM)/ S- Nitroso- N- acetyl- DL- penicillamine (10mM) i.e. PQ/SNAP was added to the cells to induce OS. Cell viability was assessed via MTS (3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium) assay while ROS produc on was determined by Flow Cytometry using the reagent CM-H2DC-FDA. The addi on of PQ/SNAP significantly reduced cell viability and increased ROS produc on (compared to un- supplemented cells, confirming the induc on of OS. Cells cultured in medium containing both 100nM and 200nM sodium selenite were significantly less affected by OS. Supplementa on with 100nM sodium selenite resulted in greater cell viability and superior quenching of ROS produc on compared to 200nM sodium selenite. The results from this in-vitro study indicate that selenium supplementa on may be effec ve in reducing the nega ve effects of OS and provides preliminary data to support the poten al role for selenium supplementa on in the preven on of CVD in overweight/obese individuals.
1st Annual meeting of SSC 2014 26 SSC Scottish Society of Cytomics
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FORESTERHILL
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1st Annual meeting of SSC 2014 28 Aberdeen Map and loca ons Please visit also follwing website for any accomoda on informa on: h p://www.visitaberdeen.com/your-visit/ Schuh Sky Superdrug Timpson Trespass Primark Quiz M&S Magnolia Menkind Miss Selfridge Monsoon Accessorize Monsoon Children New Look Next O2 Oasis Ortak Pepperberry Phase Eight Phones 4 U Radley River Island Sky Superdrug Swarovski The Body Shop The Card Factory The Perfume Shop Thorntons Topman Topshop Vision Express Vodaphone Wallis Warehouse Warren James Watch Workshop WH Smith Scan Me Aberdeen to download the the download to
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1st Annual meeting of SSC 2014 29 Aberdeen Map and loca ons
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C E O E A O C R S
R G Y L E E X D Park O A L T
Childrens Corner A F E T D RO
R R I Y O I R R H A E
I M A D O H T RO R G M E A D SINCLAIR Golf Course U D O A R L Gordon R R Y R Y S
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IL D LL F D S R B T
E K E O O A T R B W R A T D
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and Aviary C N F C ON
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I D R L W H R Winter Gardens S A
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E Q E CAIRNGORM DRIV S L N Robert Gordon’s University D L L U RT H L E T A O ON INCHGART TOL E S S R R LO IV W F D HILL DR E R D S Y A CU E L O N 46 W R R E V IG Y R G A O NU LT I W E A W E D D A E V S A R A E U D I A O O LV H R D R N T IL U K S HA E H SO N R KE Y RE R CAIRNLEE AV BA E Y A NE O S N SD W SS A IR D IE RO D LO R CA G AD H D A IG IL OA G N L IN R H AR D E D C A A AD E A D E RO S V D L SID I O M A R E R R U O E T D O L M R A IN D R E E D O U Z D O REDMOSS H E I SI S D R E T S E P SLE
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2 10 18 26 34 42
3 11 19 27 35 43 4 12 20 28 36 44
5 13 21 29 37 45 6 14 22 30 38 46
7 15 23 31 39 City map supplied by
Hotels A to Z A to Hotels 8 16 24 32 40
1st Annual meeting of SSC 2014 30 SSC Conference Commi ee
Raif Yuecel (Chairman) Linda Duncan (Deputy Secretary) IFCC, IMS IFCC, IMS University of Aberdeen University of Aberdeen [email protected] [email protected]
Mara Rocchi (Secretary) Ian Dimmick (Scien fic Advisor) Moredun Ins tute, Edinburgh Flow Cytometry Core Facility [email protected] Newcastle upon Tyne University [email protected] David Wilson (Treasurer) NHS Grampian Lynne Lumsden (Corporate Administrator) [email protected] IFCC, IMS University of Aberdeen Janice Forsyth (Event Coordinator) [email protected] IFCC, IMS University of Aberdeen [email protected]
1st Annual meeting of SSC 2014 31 Friday 26 September 2014 Flow Cytometry Data Analysis Workshop Come and learn how to save time turning your flow data into results with FCS Express software. • Easy to use - familiar Microsoft Powerpoint like interface • Publication quality graphics • Create PowerPoints and PDFs • Batch Processing • Heatmaps, bar charts & scatter plots • Powerful and flexible statistics • Cell Cycle and Proliferation • Import Accuri CFlow files automatically • Import FACS DIVA experiments • Also New FCS Express 4 Image Cytometry • Much, much, more......
5 10
4 Iain Fraser Cytometry Centre 10 CD 3+ 24.04 The Ins tute of Medical Sciences
3 10 Level 7 Conference Room
CD13+33 PE-A RSVP to [email protected] 2 -10
2 2 3 4 5 9:00am – 12:00pm -105 10 10 10 10 10 CD3 APC-A 4 Please bring any Ques ons! 10 Hope to see you there
3 10 Presented by: Andrea Valle CD13+33 PE-A 2 -10 Email : [email protected]
2 2 3 4 5 -10 10 10 10 10 CD3 APC-A
1st Annual meeting of SSC 2014 32 1st Annual meeting of SSC 2014