New XI CONGRÉS · 2017. 11. 27. · Member: Dr. Jordi Bas Minguet Member: Dra. Virgínia Mas Bosch...

XI CONGRÉS

Societat Catalana d’Immunologia (SCI)

Programa Final

Barcelona, 16 i 17 de Novembre de 2017

Regulació de la Resposta Immunitària

Immunoregulation

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Organization Committee (SCI Board):

Welcome to the Congress,

On behalf of the organising committee, we would like to warmly welcome you to the

XIth Societat Catalana d’Immunologia Congress (SCI congress). We believe that our

meeting will present high level scientific knowledge with the contribution of immunologists

and specialists who are experts in this field.

Dr. Ricardo Pujol Borrell

SCI President

XIth Congress of the Catalan Society of Immunology: Immunoregulation, has been accredited by the Catalan Lifelong Learning Board of the

Healthcare Professions with 0,7 credits (Record: 09/020427-MD, 2017.11.23)

President: Dr. Ricardo Pujol Borrell Vice President: Dra. Annabel Valledor Fernández Secretary: Dr. Francesc Rudilla Salvador Treasurer: Dra. Milagros García Ormaechea Member: Dra. Aina Teniente Serra Member: Dr. Jordi Bas Minguet Member: Dra. Virgínia Mas Bosch Member: Dra. M. Esther Moga Naranjo

Congress Office: Sr.Xavier Nieves ([email protected])

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http://www.sci.cat/

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This Congress is sponsored by:

Gold Sponsors

Silver Sponsor

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Content

Regulació de la Resposta Immunitària ................................................................................ 1

Immunoregulation ................................................................................................................ 1

This Congress is sponsored by: ........................................................................................... 4

Content ................................................................................................................................ 5

Scheme first day .................................................................................................................. 6

Scheme second day ............................................................................................................ 7

Corporative Abbreviations .................................................................................................... 9

Abstracts ............................................................................................................................ 10

Oral Communications Session I Clinical Immunology 1 - 6....................................... 10

Oral Communications Session II Innate Response 7 - 9 .......................................... 16

Oral Communications Session III From Innate to Adaptative Immunity 10 - 15 ........ 19

e-poster List ....................................................................................................................... 25

Posters Clinical Immunology 1 - 8 .................................................................................. 25

Posters Innate Response 9 - 16 ..................................................................................... 33

Posters From Innate to Adaptative Immunity 17 - 24 ..................................................... 41

2018 Events ....................................................................................................................... 49

Lifelong Learning SCI Program 2018 ............................................................................. 49

New members .................................................................................................................... 50

Registration form to SCI ................................................................................................. 50

Participant information ....................................................................................................... 51

Useful information .......................................................................................................... 51

List of participants and authors ...................................................................................... 52

Other useful information and notes ................................................................................ 54

Awards to the best communication and to the best poster at the XI Congress SCI 2017, sponsored by SCI

This year SCI sponsors the awards for the best communication (400 €) and for the best poster (250 €) of this congress. The Chairpersons of the different sessions of the congress and the board members of the SCI will select the best oral communications presented, taking into account its scientific value and the aspects related to the presentation. The poster awarded will be chosen by the congress attendees activating the electronic vote inside the electronic panels of the posters. The results will be announced at the end of the congress.

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Scheme first day

Thursday, November 16th

15:30 16:00

Arrival, Registration and Documentation delivery

16:00 16:05

Welcome to the XIth CONGRESS of SCI Dr. Ricardo Pujol Borrell (President of SCI)

16:05 17:00

Chair: Dr. Manel Juan (Hospital Clínic)

Dra. SANDRA HERVÁS CIMA, Universidad de Navarra, Spain

“Enrichment and separate expansion of tumor-infiltrating CD8 T cells expressing PD-1 improves the efficacy of adoptive therapy”

17:00 17:30

Poster viewing – Coffee Break Posters can be viewed on the 4 electronic panels located in the Hall

17:30 18:30

Chair: Dra. Marta Vives-Pi (IGTP)

Dr. JOAN VERDAGUER Universitat de Lleida, Spain

“T and B lymphocyte cross talk in Type 1 Diabetes”

18:30 20:00

Oral Communications Session I: Clinical Immunology

Chairs: Dr. Cándido Juarez (HSCSP) and Dra. Virgínia Mas (HUBell) 18:30h Immunological effects of dimethyl fumarate in multiple sclerosis patients. María José

Mansilla et al. (oral presentation 1).

18:45h Mitotic atypical IFI patterns on HEp-2 cells and Cancer association. G. Vila-Pijoan et al.

(oral presentation 2). 19:00h Virus specific T lymphocytes expansion for adoptive immunotherapy. Marta Grau-

Vorster et al. (oral presentation 3). 19:15h The importance of assessing own reference values. Sergio Navarro Velázquez et al.

(oral presentation 4). 19:30h Antigen-Specific Liposomal Nanotherapy Drives Immunoregulation in Human Type 1

Diabetes. Silvia Rodríguez-Fernández et al. (oral presentation 5). 19:45h Congenital disorder of glycosylation type II b: description of a clinical case. Andrés

Baucells de la Peña et al. (oral presentation 6).

20:00 End of session

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Scheme second day

Friday, November 17th

08:30 08:55

Arrival, Registration and Documentation delivery

09:00 10:00

Chair: Dra. Eva Martínez-Cáceres (HUGTiP)

Dr. JOSEP DALMAU IDIBAPS, UB, H. Clínic, Barcelona.

“Autoimmune encephalitis: from symptom to synapse”

10:00 11:00

Oral Communications Session II: Innate Response

Chair: Dra. Maria Rosa Sarrias (IGTP) and Dra. Annabel Valledor (UB)

10:00h Involvement of autophagy in the processing and/or presentation of diabetogenic

autoantigens by dendritic cells. Javier Vega Ramos et al. (oral presentation 7).

10:20h Mesenchymal Stem Cells induce de novo expression of CD73 in human monocytes in

vitro and in a swine model of myocardial infarction in vivo. Marta Monguió-Tortajada et

al. (oral presentation 8).

10:40h Mitofusin 2 links mitochondria with immune response. J. Tur et al. (oral presentation 9).

11:00 11:30


11:30 12:30

Chair: Dr. Pablo Engel (UB)

Dra. BELÉN DE ANDRÉS Centro Nacional de Microbiología, ISCIII, Madrid, Spain.

“Altered distribution of aged B lymphocytes. Implications on the humoral responses”

12:30 13:30

Ordinary General Meeting / Junta General Ordinària SOCIETAT CATALANA d’IMMUNOLOGIA (SCI) (12:30h – First Call) Us hi esperem a tots: els socis i no-socis!!

13:30 15:00

Poster viewing – LUNCH Posters can be viewed on the 4 electronic panels located in the Hall

15:00 16:30

Oral Communications Session III: From Innate to Adaptative Immunity

Chair: Dr. Ramón Gimeno (IMIM) and Dra. Milagros García (H. Clínic)

15:00h HLA-DRB1*15:01 and HLA-DRB5*01:01 Present Complementary Peptide Repertoires.

Erika Scholz et al. (oral presentation 10).

15:15h Role of intermediate proteasome β5i in the generation of HLA class I ligands. Erika

Scholz et al. (oral presentation 11).

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15:30h Identification of new CYCLIN D3 signaling pathways responsible for pancreatic beta cell

fitness. Celeste Santos-Rosendo et al. (oral presentation 12).

15:45h Severe phenotype of LRBA deficiency in a patient with a novel homozygous mutation

due to a chromosome 4 segmental uniparental isodisomy. Pere Soler-Palacín et al.

(oral presentation 13).

16:00h Functional study of tumor infiltrating lymphocytes in a breast cancer patient: an

approach to personalized medicine. Andrea Aran et al. (oral presentation 14).

16:15h MSC induction of Breg is mediated by secreted soluble factors but not by extracellular

vesicles. Laura Carreras-Planella et al. (oral presentation 15).

16:30 17:00


17:00 18:00

Chair: Dra. Elisabet Calderón (Instituto Grifols)

Dr. SIMON FILLATREAU INSERM, CNRS, Institut Necker-Enfants Malades (INEM), Paris, France

“Cytokine-producing B cells and plasma cells: novel key

players in immune regulation”

18:00 18:30

Chair: Dr. Ricardo Pujol Borrell (HUVH, UAB)

Drs. SIMON FILLATREAU, BELÉN DE ANDRÉS, JOAN VERDAGUER

ROUND TABLE: “B cell pathology”

18:30 18:45

Prize to the best communication and poster in the Congress, sponsored by Miltenyi Biotec and Werfen Dr. Ricardo Pujol Borrell (President of SCI). End of Congress

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Corporative Abbreviations

BST Banc de Sang i Teixits

CIBERCV Centro de Investigación Biomédica en Red Enfermedades

Cardiovasculares

CIBERDEM Centro de Investigación Biomédica en Red de Diabetes y

Enfermedades Metabólicas Asociadas

CIBERehd Centro de Investigación Biomédica en Red de Enfermedades

Hepáticas y Digestivas

CIG, UNIL Center fo Integrative Genomics, Université de Lausanne

CIMA Centro de Investigación Médica Aplicada

HSCSP Hospital de la Santa Creu i Sant Pau

HUBell Hospital Universitari de Bellvitge

HUGTiP Hospital Universitari Germans Trias i Pujol

HUVH Hospital Universitari Vall d’Hebron

ICREA Institució Catalana de Recerca i Estudis Avançats

ICREC Insuficiència Cardíaca i REgeneració Cardíaca

IDIBAPS Institut d'Investigacions Biomèdiques August Pi i Sunyer

IDIBELL Institut d'Investigació Biomèdica de Bellvitge

IGTP Institut d’Investigació en Ciències de la Salut Germans Trias i Pujol

IIBB Institut d'Investigacions Biomèdiques de Barcelona

IMIM Institut Hospital del Mar d'Investigacions Mèdiques

INSA Institut de Recerca en Nutrició i Seguretat Alimentària

INSERM Institut National de la Santé Et de la Recherche Médicale

IRB Barcelona Institut de Recerca Biomèdica Barcelona

IRB Lleida Institut de Recerca Biomèdica Lleida

ISCiii Instituto de Salud Carlos III

PCB Parc Científic de Barcelona

REMAR-IVECAT REcerca en Malalties d’Afectació Renal- Innovation in Vesicles and

Cells for Application in Therapy

SCI Societat Catalana d’Immunologia

UAB Universitat Autònoma de Barcelona

UB Universitat de Barcelona

UPF Universitat Pompeu Fabra

VHIR Vall d’Hebron Institut de Recerca

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Abstracts

Oral Communications Session I

Clinical Immunology 1 - 6

1 Immunological effects of dimethyl fumarate in multiple

sclerosis patients

María José Mansilla1,2

; Juan Navarro-Barriuso1,2

; Silvia Presas-Rodríguez3,4

; Aina Teniente-Serra1,2

;

Bibiana Quirant-Sánchez3,4

; Cristina Ramo-Tello3,4

; Eva M. Martínez-Cáceres1,2

1Immunology Division, Germans Trias i Pujol University Hospital and Research Institute.;

2Department of

Cellular Biology, Physiology and Immunology, Universitat Autònoma de Barcelona; 3Multiple Sclerosis Unit,

Department of Neurosciences, Hospital Universitari Germans Trias i Pujol; 4Department of Medicine,

Universitat Autònoma de Barcelona

Background: Dimethyl fumarate (DMF) is an oral drug for the treatment of relapsing-

remitting multiple sclerosis (RRMS), with an anti-inflammatory effect. It is relatively well

tolerated, but it has an important impact over several leukocyte subpopulations.

Consequently, immune monitorization becomes necessary to understand the effect of

DMF on the immune system and to relate these changes to the clinical outcome and

potential adverse effects.

Objective: To analyze the immunological changes induced by DMF in samples of whole

blood of RRMS patients.

Methods: Longitudinal prospective study of peripheral blood T, B, NK, monocyte and DC

subpopulations using multiparametric flow cytometry in whole blood from 11 RRMS

patients under DMF treatmentat baseline and after 1, 3, 6 and 12 months of follow up.

Results: The study revealed a selective reduction of effector central memory T cells and

memory B cells in a 52.3±14.1%, 37.3±16.1% and 54.3±20.3%, respectively after 12

months. In contrast, the relative prevalence of both T and B naïve subpopulations

increased in a 58.4±25.6% and 47.1±31.6%, respectively. Additionally, a switch in the

central memory Th2/Th1 ratio from 0.8±0.3 to 1.4±0.6 and an increase of transitional B

cells in absolute number (from 5±3 to 10±5 cells/μL) were spotted. All these changes were

progressive from month 1, and already significant by month 6. All results were statistically

significant (p<0.05).

Conclusions: The beneficial effect of DMF reducing the number of clinical relapses in MS

patients seems to be related with the effect of DMF depleting effector T cells and

increasing transitional B cells.

This work was partially supported by project PI14/01175, integrated in the Plan Nacional

de I+D+I and cosupported by the ISCIII-Subdirección General de Evaluación and the

Fondo Europeo de Desarrollo Regional (FEDER).

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2 Mitotic atypical IFI patterns on HEp-2 cells and Cancer

association

Vila-Pijoan G.

1; Sanz-Martínez M.T.

1; Pujol-Borrell R.

1,2; Marín-Sánchez A.

1

1Immunology Division, Hospital Universitari Vall d’Hebron (HUVH), Barcelona, Catalonia, Spain;

2Immunology Group, Vall d’Hebron Research Institute (VHIR), Barcelona, Catalonia, Spain

Introduction: An association between autoimmune diseases and cancer is well

established, and it has been in part attributed to the immune-system response to cancer

antigens. With the advent of immune-checkpoint immunotherapy many cancer patients are

tested for autoantibodies prior to treatment and this constitutes an opportunity to

investigate other tumor-associated autoantibodies.

Material and Methods: We selected the cases corresponding to ANA tests on HEp-2 cells

(NOVA Lite®, INOVA Diagnostics, Werfen) reported as "mitotic atypical pattern" by our

laboratory among all the samples tested from 2015 to 2017. Positive sera were classified

following the International Consensus on ANA patterns (ICAP) as CENP-F-like (AC-14),

mitotic chromosomal coat (AC-28); we also considered a group that showed the so called

“Pleomorphic-not-PCNA” pattern. Clinical records were reviewed for cancer diagnosis and

the results of tumor markers (CEA, hCG, CA 125, CA 19.9, CA 15.3, CA 72.4, SCC

antigen, B2MG, AFP, enolase, CYFRA 21,1, TK) if available.

Results: From 36.541 ANA tests, 44 from 22 patients were reported as “mitotic atypical

pattern”: (13 Pleomorphic-not-PCNA, 7 CENP-F-like and 2 mitotic chromosomal coat).

Clinical records revealed that 5/22 had a cancer diagnosis, 4/22 showed the CENP-F-like

pattern and 1/22 the Pleomorphic-not-PCNANop- 52-like. Tumor markers were available in

9/22 patients and were positive in 5 of whom, 2 showed a mitotic chromosomal coat

pattern without a malignancy diagnosis at the time of the study.

Discussion: We have confirmed that the CENP-F-like pattern is strongly associated to

malignancy (57% of the cases in our study). Interestingly, the two patients positive for the

mitotic chromosomal coat pattern had elevated laboratory tumor markers.

Conclusions: Even though further studies are needed to elucidate whether they can be

used as biomarkers, the detection of atypical patterns in ANA screening test on HEp-2

cells should alert the clinicians to the possibility of a malignancy.

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3 Virus specific T lymphocytes expansion for adoptive

immunotherapy

Marta Grau-Vorster1,2

; Anna del Mazo-Barbara1; María López-Montañés

1,2; Sergi Querol

1; Joaquim

Vives1; Irene Oliver-Vila

1; Francesc Rudilla

1

1Banc de Sang i Teixits (BST);

2Vall d'Hebron Institut de Recerca (VHIR)

Introduction: Adoptive immunotherapy with virus-specific T lymphocytes (VST) can

prevent immunodeficient patients from virus reactivation or de novo infection by

reconstituting the immune system. Nowadays, this treatment has several limitations, such

as HLA-compatibility restriction, and the high costs and time required in the production

processes of personalised medicines.

Experimental approach: We propose the generation of a third-party VST bank through

large-scale expansion compliant with Good Manufacturing Practice standards while

reducing production costs and time, and with its pertinent HLAmatched virus-specific

register.

Methodology: A full factorial design of experiments (DoE) was used for the screening of

different cytokine combinations (IL-2, IL-7, IL-12 and IL-15) in VST media formulation.

Traditional plastic culture system and siliconemembrane technology (G-Rex, Wilson Wolf

Manufacturing) were also compared. First, cytomegalovirus (CMV) peptides were used to

stimulate lymphocytes specifically. Readouts included purity, in terms of CD8+ and CD4+,

and specificity (interferon gamma (IFN-γ) expression). This strategy was compared to

other methods found in the literature that also used peptides for specific stimulation. Once

established the optimal media composition, a physiological VST stimulus was tested,

consisting of viral peptide-pulsed dendritic cells (DCs) derived from monocytes.

Furthermore, anti-CD3+anti-CD28 antibodies were added after 5 days of culture,

according to the latent period, in order to enhance VST proliferation.

Results: The combination of the cytokines IL-2+IL-7+IL-15 in a G-Rex system resulted in

optimum conditions for VST expansion. The use of DCs for T lymphocyte stimulation and

the addition of anti-CD3+anti-CD28 resulted in a successful 14-day expansion with high

expansion factor (80-fold) and specificity (16.4%).

Conclusions: A scalable process with high values of VST was successfully defined by

formulating cell culture media with IL-2+IL-7+IL-15, stimulating lymphocytes with pulsed-

DCs and enhancing cell growth by adding anti-CD3+anti-CD28 5 days later. A further

improvement to the approach presented here would consider specificity for other viruses.

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4 The importance of assessing own reference values

Sergio Navarro Velazquez1; Virgínia Mas Bosch

1; Francisco Morandeira Rego

1; Elisabet Poyatos

Canton1; Rafael Mayol Calvet

1; Jordi Bas Minguet

1

1Department of Immunology, Bellvitge University Hospital

Introduction: Many clinical laboratories use reference values established by the

manufacturer but these values may not always apply to their target populations.

Objectives: Therefore, we attempted to establish our own reference values for different

serum proteins: complement factor 3 (C3c), complement factor 4 (C4), complement factor

B or properdin (PFB), ceruloplasmin (CER), alpha-1-antitrypsin (AAT) and complement

factor 1 inhibitor (C1I).

Materials and methods: Blood samples were obtained from 50 healthy individuals (58%

women): 25 blood donors and 25 lab staff volunteers after completion of a health

questionnaire. 50% were fasting. Samples were centrifuged (4000 rpm for 10’) and sera

were frozen at -80° C until analysis. The concentration of the different proteins was

determined by nephelometry (BNII, Siemens®) in 5 different batches. A statistical study

with analyze-it® was carried out, which showed that fasting did not affect the results of any

test. After removing the outlier values, Harris and Boid partition criteria for age and gender

were applied. Then, Poison’s distribution tests were performed and reference intervals

were calculated.

Results: All values followed a normal (or normal-logarithmic) distribution. The newly

obtained reference values were clearly different for CER (Before 220-610 mg/L, After 178-

431 mg/L), but also in the other lab tests, albeit to a lesser extent. Fasting and age had no

influence on these values; however, gender clearly influenced the reference values of CER

(Women 201-465 mg/L, Men 167-381 mg/L) and AAT (Women 951-1785 mg/L, Men 861-

1591 mg/L), recommending the establishment of gender-specific reference values for

these tests.

Conclusions: It would be appropriate to validate these reference intervals with a further

sample batch before setting the new reference values. Furthermore, it is highly

recommended to carry out similar studies in all laboratories to assess whether the

reference values proposed by manufacturers are appropriate in their own area of

influence.

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5 Antigen-Specific Liposomal Nanotherapy Drives

Immunoregulation in Human Type 1 Diabetes

Silvia Rodríguez-Fernández

1; Irma Pujol-Autonell

1; Ferran Briansó

2,3; David Perna-Barrull

1; Mary

Cano-Sarabia4; Sonia García-Jimeno

4; Adrian Villalba

1; Alex Sánchez

2,3; Eva Aguilera

5; Federico

Vázquez5; Joan Verdaguer

6,7; Daniel Maspoch

4,8; Marta Vives-Pi

1,7

1Immunology Division, Germans Trias i Pujol Research Institute, UAB, Badalona;

2Statistics and

Bioinformatics Unit, Vall d'Hebron Research Institute, Barcelona; 3Department of Genetics, Microbiology and

Statistics, University of Barcelona, Barcelona; 4Catalan Institute of Nanoscience and Nanotechnology,

Bellaterra; 5Endocrinology Section, Germans Trias i Pujol University Hospital, Badalona;

6Department of

Experimental Medicine, University of Lleida, IRBLleida, Lleida; 7CIBERDEM, ISCiii, Barcelona;

8ICREA,

Barcelona

Type 1 diabetes (T1D) is a metabolic disease caused by the autoimmune destruction of

insulin-producing pancreatic beta-cells, which, to date, cannot be prevented or reverted.

New and antigen-specific strategies are necessary to re-establish self-tolerance and allow

target tissue regeneration. Relying on the tolerogenic potential of apoptotic cells, we

demonstrated that liposomes mimicking apoptotic beta-cells can halt the autoimmune

attack and prevent T1D in a mouse model of the disease through the generation of

tolerogenic dendritic cells (DCs). These liposomes are rich in phosphatidylserine (PS) —a

characteristic phospholipid of the apoptotic cell membrane— and contain beta-cell

autoantigens. To validate the tolerogenic effect of apoptotic mimicry in human T1D, PS-

liposomes loaded with human insulin peptides as autoantigens were generated with

optimum size and composition for phagocytosis. DCs were derived in vitro from peripheral

blood monocytes isolated from patients with T1D and control age-related subjects, with

similar yield, purity, viability and efficiency of differentiation. Human DCs from both groups

rapidly and efficiently phagocyted PS-liposomes with identical phagocytosis kinetics.

Engulfment of PS-liposomes induced a tolerogenic phenotype and functionality in DCs,

which displayed low membrane expression of class I and II HLA, CD40, CD86 and CD25

molecules and impaired capacity to induce autologous T cell proliferation. RNAsequencing

was performed in DCs to detect changes after PS-liposomes phagocytosis, showing 233

differentially expressed genes primarily taking part in immune response, cytokine signaling

and vesicle transport. Genes involved in antigen presentation were downregulated

whereas tolerogenic-related genes were mostly upregulated. In conclusion, PS-liposomes

phagocytosis leads to phenotypic and functional changes in human DCs, analogous to

those observed in mice and accountable for tolerance induction and autoimmunity

blockade. These results reinforce the potential of this immunotherapy to re-establish

immunological tolerance, opening the door to new approaches in autoimmunity.

Funding: ISCiii (PI15/00198); CERCA and AGAUR, GenCat; La Marató de TV3 (201632-

10).

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6 Congenital disorder of glycosylation type II b: description of a

clinical case

Andrés Baucells de la Peña

1; Gemma Boera-Carnicero

1; Aida Pariente-Cano

1; Isabel Badell

2; Oscar

de la Calle-Martín1; Laura Martínez-Martínez

1

1Immunology Dept., Hospital de la Santa Creu i Sant Pau;

2Pediatrics Dept., Hospital de la Santa Creu i Sant

Pau

Introduction: The congenital disorders of glycosylation (CDGs) are genetic disorders

affecting the Nglycosylation process. Alterations in the endoplasmic reticulum glucosidase

I, the mannosyl-oligosaccharide glucosidase (MOGS) encoded by the MOGS gene, are

responsible for CDG type IIb (CDG-IIb). Only 3 patients worldwide have been identified

with this extremely rare disease. The first one, described in 2000, was a neonate with

severe generalized hypotonia, hypoventilation, dysmorphic features, hepatomegaly,

feeding problems and seizures. He died at age 74 days. The other 2 patients were

described in 2014 and were two siblings of 11 and 6 years old with dysmorphic features

affected by a complex neurological condition including hypotonia and seizures. Both

patients presented with severe hypogammaglobulinemia but curiously, without remarkable

infections.

Material and methods: We report a 15 months old boy who was remitted to study due to

severe hypogammaglobulinemia (IgG:164 mg/dL, IgA:<7 mg/dL, IgM:16 mg/dL). The

patient was a premature neonate (29 week, 1530 g) with failure to thrive. He also

presented with anisocoria and feeding problems. His clinical history of infections was

limited to recurrent enterovirus constipations and intermittent bronchitis. First, X-linked

agammaglobulinemia was suspected. But the presence of high lymphocyte numbers,

including B cells (34.8%), together with dysmorphic features (macrocephaly, bulging

forehead, low-set ears and prominent abdomen) prompted us to suspect a CDG-IIb. So,

MOGS gene was analysed by sequencing.

Results: We identified two diferent heterozygotic mutations in the MOGS gene: c.G715A,

resulting in the substitution of aspartic 239 by asparragine in the maternal allele, and

c.C877T changing proline 293 by serine in the paternal allele. Both parents were

heterozygous for each of these mutations. None of them were found in the three previous

reported patients.

Conclusion: Severe hypogammaglobulimenia together with dysmorphic features led us to

identify a new patient with CDG-IIb bearing two novel mutations: p.Asp239Asn and

p.Pro293Ser.

16 www.sci.cat

Oral Communications Session II Innate Response 7 - 9

7 Autophagic machinery in DCs plays a critical role in

diabetogenesis by impairing CD4 T cell activation and

recruitment Javier Vega Ramos

1; Elena López

1; Pau Serra

1; Pere Santamaria

1,2

1Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) Barcelona, Spain;

2Julia McFarlane

Diabetes Research Centre and Department of Microbiology, Immunology and Infectious Diseases, Cuming

School of Medicine, University of Calgary, Canada

Atg5 and other autophagy related proteins have been found to be necessary for optimal

processing and presentation of phagocytosed antigens by conventional Dendritic Cells

(DC), but the role of this process in the activation of diabetogenic CD4 and/or CD8 T-cells

is unknown. Currently, we are investigating the role of the autophagy machinery (Atg5) in

DC for direct priming of diabetogenic CD4 and CD8 T cells. To address this question, we

generated mice expressing CD11c- driven Cre transgenes in non-transgenic, CD4 TCR

transgenic (4.1) and CD8 TCR transgenic (8.3), NOD.Atg5loxP/loxP. In these mice,

autophagic machinery is knocked down specifically in DC. We found that the incidence of

diabetes is clearly reduced in non-transgenic and 4.1-TCR transgenic mice, showing an

important role for autophagy in the MHC-II driven antigen processing and/or presentation

of diabetogenic autoantigens to CD4 T cells in vivo. On the other hand, 8.3-TCR

transgenic mice only showed a minor reduction in the incidence of diabetes, showing that

ATG5 deletion in DC does not impair cross-presentation of diabetogenic autoantigens to

8.3-CD8 T cells, nor 8.3-CD8 T cells recruitment and activation. No phenotypical

differences have been observed in T cells from CD11c-Cre expressing 4.1 or 8.3-TCR

transgenic NOD.Atg5loxP/loxP mice and their control counterparts. Altogether, our data

suggest that the autophagic machinery in DC plays a critical role in the activation of

diabetogenic CD4 T cells, presumably via effects on exogenous antigen presentation of

diabetogenic autoantigens.

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Oral Communications Session II

Innate Response 7 - 9

8 Mesenchymal Stem Cells induce de novo expression of CD73

in human monocytes in vitro and in a swine model of

myocardial infarction in vivo Marta Monguió-Tortajada

1,2; Santiago Roura

3,4,5; Carolina Gálvez-Montón

3,5; Marcella Franquesa

1,6;

Antoni Bayes-Genis3,5,7,8

; Francesc E. Borràs1,2,6

1REMAR-IVECAT Group, Health Science Research Institute Germans Trias i Pujol, Can Ruti Campus;

2Universitat Autònoma de Barcelona;

3ICREC Research Program, Health Science Research Institute

Germans Trias i Pujol, Can Ruti Campus; 4Center of Regenerative Medicine in Barcelona;

5CIBERCV,

Instituto de Salud Carlos III; 6Nephrology Service, Germans Trias i Pujol University Hospital;

7Cardiology

Service, Germans Trias i Pujol University Hospital; 8Department of Medicine, UAB

The ectoenzymes CD39 and CD73 regulate the purinergic signaling through thehydrolysis

of ATP/ADP to AMP and to Adenosine, respectively. This shifts the pro-inflammatory

milieu induced by extracellular ATP to the anti-inflammatory regulation by Adenosine.

Mesenchymal stem cells (MSCs) have potent immunomodulatory capabilities, including

monocyte modulation towards an anti-inflammatory phenotype aiding tissue repair. In vitro,

we observed that human cardiac adipose tissue-derived MSCs (cATMSCs) and umbilical

cord MSCs (UCMSCs) similarly polarize monocytes towards a regulatory M2 phenotype,

which maintained the expression of CD39 and induced expression of CD73 in a cell

contact dependent fashion, correlating with increased functional activity.

In addition, the local treatment with porcine cATMSCs using an engineered bioactive graft

promoted the in vivo CD73 expression on host monocytes in a preclinical swine model of

myocardial infarction.

Our results suggest the upregulation of ectonucleotidases on MSCconditioned monocytes

as an effective mechanism to amplify the long-lasting immunomodulatory and healing

effects of MSCs delivery.

18 www.sci.cat

Oral Communications Session II


9 Mitofusin 2 links mitochondria with immune response

Juan Tur1; Selma Pereira-Lopes

1; Tania Vico

1; Maribel Hernández-Alvarez

2,3,4; Juan P. Muñoz

2,3,4; Emil

R. Unanue 5; Antonio Zorzano

2,3,4; Antonio Celada

1; Jorge Lloberas

1

1Grup Biologia del Macròfag, Departament de Biologia Cel·lular, Fisiologia i Immunologia, PCB, UB;

2Centro

de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM; 3Institute for Research in Biomedicine (IRB Barcelona);

4Departament de Bioquímica i Biomedicina

Molecular, Facultat de Biologia, UB; 5Department of Pathology and Immunology; Washington University

School of Medicine in St. Louis

Mitofusin 2 (Mfn2) plays a major role in mitochondrial fusion and the establishment of

mitochondriaendoplasmic reticulum contact sites. Mitochondria are critical regulators of the

metabolism, and have a central role in the innate immune system. Because macrophages

play a major protagonist in inflammation, here we analyze the effect of Mfn2 on the

functional activity of these cells. Using a Mfn2 myeloid conditional KO mouse we show that

in macrophages, Mfn2 is required for the adaptation of mitochondrial respiration to stress

conditions, and for the production of reactive oxygen species (ROS) upon proinflammatory

activation. The lack of ROS production is at the base of a defective production of

proinflammatory cytokines and nitric oxide. In addition, the lack of Mfn2 is associated to a

dysfunctional autophagy, apoptosis, phagocytosis, and antigen processing. These results

unravel an unexpected role for Mfn2 in macrophages, involving ROS production, and

inflammation.

www.sci.cat 19

Oral Communications Session III From Innate to Adaptative Immunity 10 - 15

10 HLA-DRB1*15:01 and HLA-DRB5*01:01 Present

Complementary Peptide Repertoires

Erika Scholz1,2

; Miguel Marcilla3; Xavier Daura

1,4; David Arribas-Layton

5; Eddie A. James

5; Iñaki

Álvarez1,2

1Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Spain;

2Immunology Unit, Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de

Barcelona; 3Proteomics Unit, Centro Nacional de Biotecnología (CSIC), Madrid, Spain;

4Catalan Institution

for Research and Advanced Studies (ICREA), Barcelona, Spain; 5Benaroya Research Institute at Virginia

Mason, Seattle, WA, United States

Human leukocyte antigen (HLA)-DR15 is a haplotype associated with multiple sclerosis. It

contains the two DRB* genes DRB1*1501 (DR2b) and DRB5*0101 (DR2a). The reported

anchor motif of the corresponding HLA-DR molecules was determined in 1994 based on a

small number of peptide ligands and binding assays. DR2a could display a set of peptides

complementary to that presented by DR2b or, alternatively, a similar peptide repertoire but

recognized in a different manner by T cells. It is known that DR2a and DR2b share some

peptide ligands, although the degree of similarity of their associated peptidomes remains

unclear. In addition, the contribution of each molecule to the global peptide repertoire

presented by the HLA-DR15 haplotype has not been evaluated. We used mass

spectrometry to analyze the peptide pools bound to DR2a and DR2b, identifying 169 and

555 unique peptide ligands of DR2a and DR2b, respectively. The analysis of these sets of

peptides allowed the refinement of the corresponding binding motifs revealing novel

anchor residues that had been overlooked in previous analyses. Moreover, the number of

shared ligands between both molecules was low, indicating that DR2a and DR2b present

complementary peptide repertoires to T cells. Finally, our analysis suggests that,

quantitatively, both molecules contribute to the peptide repertoire presented by cells

expressing the HLA-DR15 haplotype.

20 www.sci.cat

Oral Communications Session III From Innate to Adaptative Immunity 10 - 15

11 Role of intermediate proteasome β5i in the generation of HLA

class I ligands

Erika Scholz1; Anna Mestre-Ferrer

1; Françesc Canals

2; Montserrat Carrascal

3; Iñaki Álvarez

1

1Immunology Unit, Department of Cell Biology, Physiology and Immunology and Institut de Biotecnologia;

2Vall d'Hebron University Hospital Research Institute Proteomics Laboratory, Barcelona, Spain;

3CSIC/UAB

Proteomics Laboratory, IIBB-CSIC/Institut d'Investigacions Biomèdiques August Pi i Sunyer

The proteasome is a multicatalytic complex responsible for most of the cytosolic protein

degradation. In addition to the standard proteasome (containing the catalytic subunits β1,

β2 and β5) and the immunoproteasome (β1i, β2i and β5i), intermediate proteasomes are

present in normal tissues and, in a higher percentage, in dendritic and tumoral cells. Two

intermediate proteasomes have been described: one containing β5i with the components

of the standard proteasome β1 and β2, and other containing the components of the

immunoproteasome β1i and β5i with β2. Their relevance has been demonstrated as some

HLA-I ligands derived from tumors are generated specifically by intermediate

proteasomes. The activity of intermediate proteasomes have been studied through

digestions with purified 20S proteasome of fluorogenic substrates or HLA-I ligand peptide

precursors. Here, we identified by the first time some peptides associated to HLA-I in cells

expressing the constitutive proteasome or the intermediate proteasome β5i. The

generation of some specific or common HLA-I ligands sequenced in these cells was

studied by in vitro digestions of peptide precursors with purified 20S proteasomes. The

cleavage of the peptide bonds generating the N- and C-termini of the ligands by both

proteasomes allowed to explain the presence or absence of some ligands in each cell.

Finally, an exhaustive analysis of the proteasome specificity showed that the intermediate

proteasome β5i presents an increased trypsin-like activity and a decreased chymotrypsin-

like activity.

www.sci.cat 21

Oral Communications Session III

From Innate to Adaptative Immunity 10 - 15

12 Identification of new CYCLIN D3 signaling pathways

responsible for pancreatic beta cell fitness

Celeste Santos-Rosendo1; Celia Vived

1; Isabel López Mejía

2; Estela Rosell-Mases

1; Leire Egía-

Mendikute1; Marta Corral

1; Joan Verdaguer

1; Thomas Stratmann

3; Lluís Fajas

2; Conchi Mora

1

1Immunology Unit. Departament of Experimental Medicine. University of Lleida IRBLleida;

2Center for

Integrative Genomics, CIG, UNIL Sorge, Lausanne, Switzerland; 3Department of Cell Biology, Physiology

and Immunology, Faculty of Biology, University of Barcelona.

Autoimmune diabetes (T1D) is caused by the destruction of insulin producing pancreatic

beta cells. Proinflammatory cytokines released in the autoimmune attack deeply alter the

physiology and viability of beta cells leading them to an apoptotic death.

Cyclin D3 is involved in CDK-dependent cell cycle progression. Our group has reported

that cyclin D3, which is downregulated in beta cells upon inflammation, is essential for

protecting beta cells in front of the inflammation-induced apoptosis and for maintaining

proper function of beta cells, both in a cell-cycle independent fashion.

We evaluated differences in the kinome from between pancreatic islets from 11-week- old

NODSCID either wild type (NS-WT) or cyclin D3 deficient (NS-KO) female mice, that had

been submitted to glucose challenge intraperitoneally after fasting for 16h. The kinome

was assessed with tyrosine and serine/threonine kinase microarrays (PamGene

International BV). It was performed according to the manufacturer’s instructions on a

PamStation12 instrument. The list of peptides the phosphorylation of which is significantly

different between control (NS-WT) and test (NS-KO) conditions were uploaded to GeneGo

for pathway analysis.

Then, the expression and activity of selected candidate proteins was assessed by Western

Blot using pancreatic islets from both, 11 week NS-KO and NS-WT female hyperglucemic

mice by glucose injection.

We focused on those differences not attributable to the CDK activity.

From the differential kinome analysis we obtained a list of candidates. We selected the

candidates that were not related with the cell cycle and proliferation. Cyclin D3 deficiency

has a dramatic impact on the activity of some the Ser/Thr kinase. Most of Ser/Thr kinase

showed more activity in NS-KO than in NS-WT islets. In the opposite sense, on average,

most of Tyr kinases were less active in the absence of cyclin D3.

22 www.sci.cat



13 Severe phenotype of LRBA deficiency in a patient with a novel

homozygous mutation due to a chromosome 4 segmental

uniparental isodisomy

Pere Soler-Palacín1,2

; Marina García-Prat1,2

; Andrea Martín-Nalda1,2

; Clara Franco-Jarava2,3

; Mónica

Martínez-Gallo2,3

; Alberto Plaja4; Mattia Bosio

5; Stephan Ossowski

5; Roger Colobran

2,3,4

1Pediatric Infectious Diseases and Immunodeficiencies Unit (UPIIP), Hospital Universitari Vall d’Hebron;

2Jeffrey Model Foundation Excellence Center, Barcelona, Catalonia, Spain.;

3Immunology Division, Hospital

Universitari Vall d’Hebron (HUVH), Vall d’Hebron Research Institute (VHIR); 4Department of Clinical and

Molecular Genetics, Hospital Universitari Vall d’Hebron (HUVH), Barcelona; 5Centre for Genomic Regulation

(CRG), The Barcelona Institute of Science and Technology, and Universitat Pompeu Fabra (UPF),

Barcelona, Catalonia, Spain

LRBA deficiency was described for the first time in 2012 as an autosomal recessive

disorder caused by biallelic mutations in LRBA gene (OMIM #614700). It was initially

characterized by early-onset hypogammaglobulinemia, autoimmune manifestations,

susceptibility to inflammatory bowel disease (IBD), and recurrent infections. However,

further reports clearly expanded this phenotype (including patients without

hypogammaglobulinemia) showing that LRBA deficiency is a clinically variable syndrome

with a wide spectrum of clinical manifestations. We present a female patient who at the

age of 8 months presented with type 1 diabetes, psoriasis, oral thrush and enlarged liver

and spleen. She also suffered recurrent bacterial and viral infections including

pneumococcal meningitis and Epstein Barr (EBV) viremia. She underwent two consecutive

stem cell transplants (SCT) at the age of 8 and 9 years, and finally died.

The patient and her parents were subjected to WES (Whole Exome Sequencing), which

revealed a homozygous 1-bp insertion in exon 23 of LRBA patient’s gene causing a

truncated protein. The patient’s healthy mother was heterozygous for the mutation and her

father was wild type. This finding suggested that either one copy of paternal Chr4 bore a

deletion including the LRBA locus, or the patient inherited two copies of the mutant

maternal LRBA allele. Patient’s WES data showed a loss of heterozygosity (LOH) region

of 1 Mb in Chr4 including LRBA gene. Comparative genomic hybridization (CGH) array of

the patient and father genomic DNA was performed with normal results, ruling out genomic

copy number abnormalities.

Here we present for the first time a patient with LRBA deficiency due to a uniparental

disomy (UPD). In contrast to classical Mendelian inheritance, UPD is the inheritance of 2

copies of a region of chromosome from only one parent. Specifically, our patient carries a

small segmental isodisomy of maternal origin affecting 1 Mb of chromosome 4.

www.sci.cat 23



14 Functional study of tumor infiltrating lymphocytes in a breast

cancer patient: an approach to personalized medicine

Andrea Aran1; Mireia Bernuz

1; Vicente Peg

2; Cristina Bernadó

2; Esther Zamora

2; Jesús Soberino

3;

José Pérez3; Esther Holgado

3,4; Joaquín Arribas

2; Javier Cortés

2,3,4; Mercé Martí

1

1Institut de Biotecnologia i Biomedicina - Universitat Autònoma de Barcelona;

2Vall d’Hebron Institute of

Oncology; 3Instituto Oncológico Baselga - Quirón Hospital;

4Ramón y Cajal University Hospital

Breast cancer (BC) is the most common of women cancers. Triple-negative BC (TNBC),

negative for estrogen and progesterone receptor and HER-2 genes, represent a clinical

challenge because they do not respond to endocrine therapy or other targeted agents.

Thus, there is an urgent need of a personalized approach to treatment. Studying of tumor

infiltrating lymphocytes (TILs) is a promising field because of their good correlation with

patient survival, especially those with high CD8/Treg ratio.

We have studied TILs from a TNBC patient to characterize the immune response. TILs

were obtained from a core biopsy that was cut in serial slices and cultured. IHC was

performed to observe the infiltration of T cells in the tumor site. Molecular and functional

phenotype of TILs was studied by immunostaining of TILs, cytokine release and

suppression assays, all analyzed by flow cytometry. TCRs from TILs, before and after

expansions, have been sequenced in order to find if there are monoclonal expansions.

IHC images showed a high infiltration of CD8 and CD4 T cells. Although TILs were a

mixture of both populations in all cultures, different CD8/CD4 ratios were observed related

with their location on the biopsy. This different distribution of CTLs and CD4 TILs also

affected the immune mediators detected in the supernatant, i.e. higher presence of

cytotoxicity-related proteins Granzyme B and IFN-γ in cultures with CTLs dominance. No

cytokine profile could be defined on cultured with predominant CD4 T cells. Then, TILs

were expanded in vitro and tested in standard regulation assays. Cultures with

predominant CTLs showed less capacity of inhibiting alloreactive proliferation compared

with CD4 T cell cultures.

We have observed a heterogeneous distribution of TILs in the biopsy that may be useful to

select the appropriate T cells to design tailored approaches to TNBC treatment.

24 www.sci.cat



15 MSC induction of Breg is mediated by secreted soluble factors

but not by extracellular vesicles

Laura Carreras-Planella1,2

; Marta Monguió-Tortajada1,2

; Francesc E. Borràs 1,2,3

; Marcella Franquesa1,3

1REMAR-IVECAT Group, Germans Trias i Pujol Health Science Research Institute, Can Ruti Campus,

Badalona; 2Department of Cell Biology, Physiology and Immunology, Autonomous University of Barcelona,

Barcelona; 3Nephrology Department, Germans Trias i Pujol University Hospital, Can Ruti Campus,

Badalona, Spain.

Regulatory B cells (Breg), one of the newest members of the regulatory immune cells

family, are in the spotlight for their role in immune homeostasis maintenance and tolerance

achievement in autoimmune diseases and transplantation. Secretion of IL-10 is their

widest accepted mechanism of action. Mesenchymal stem or stromal cells (MSC) have

attracted the attention for their immunomodulatory actions on T cells,

monocytes/macrophages and NK making them promising therapeutic tools in immune-

related diseases. More recently it has been shown that MSC immunomodulate B cells by

abrogating plasmablast formation, maintaining an inactivated phenotype (CD27-) and

inducing IL-10-producing Breg.

The aim of this study is to elucidate the mechanism

by which MSC induce Breg focusing on the effect of

MSC culture supernatant (SN) on B cells. Using size

exclusion chromatography, MSC SN was

fractionated to separate extracellular vesicles (EV)

from other secreted factors, which are mostly

proteins (Fprot). Then, we tested the effect of the

whole or the fractionated SN on B cells and

analysed the proportion of the different B cell

subsets by flow cytometry and their secretion of IL-

10 by ELISA.

We showed that MSC culture SN can partially

induce B cell subsets modification and IL-10 secretion in a similar way to that of MSC.

Fprot fractions, rich in proteins and free from EVs, shaped B cell populations like MSC and

also induced IL-10 secretion, contrary to EV-containing fractions, which had no effect on B

cells. This indicates that the effect of MSC on B cells is at least partially induced by

secreted soluble molecules. When properly identified and isolated, MSC’s secreted

proteins could be used as a therapeutic agent in immune-mediated diseases taking

advantage of their capacity of Breg induction.

www.sci.cat 25

e-poster List

Posters

Clinical Immunology 1 - 8 The authors will attend the poster on 16/11/2017: 17:30h; 17/11/2017: 11:00h, 13:30h, 16:30h.

1 Wiskott-Aldrich syndrome mimicking classical combined

immunodeficiency

Perurena-Prieto, Janire1; Franco-Jarava, Clara

1,5; Mari, Ana V.

2; Domínguez-Alonso, Carmen

2; Martín-

Nalda, Andrea3,5

; Aragon, Larraitz1; García-Prat, Marina

3,5; Colobran, Roger

1,4,5; Regueiro, Jose R.

2;

Soler-Palacín Pere3,5

1Immunology Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain;

2Dept. of Immunology,

Faculty of Medicine, Complutense University, and Hospital 12 de Octubre Health; 3Pediatric Infectious

Diseases and Immunodeficiencies Unit, Hospital Universitari Vall d'Hebron, Barc; 4Genetics Department,

Hospital Universitari Vall d’Hebron, Barcelona, Spain; 5Jeffrey Modell Diagnostic and Research Center for

Primary Immunodeficiencies, Barcelona, Spain

Introduction: Wiskott-Aldrich syndrome (WAS) is a rare X-linked recessive primary

immunodeficiency characterized by micro-thrombocytopenia and immune-deregulation.

WAS is caused by mutations in the WAS gene that codes the WAS protein (WASP), a key

regulator of the actin cytoskeleton. The diagnosis of WAS can be difficult because of its

variable clinical presentation. Minor abnormalities of lymphocyte subsets and function are

frequent, but usually the lymphocyte count is normal in early childhood.

Objective: To illustrate the difficulties of immunological diagnosing of WAS in early

childhood by reporting a case recently diagnosed at our center.

Clinical data and methods: A 3-months old boy with persistent laryngo-tracheo-bronchitis

and thrombocytopenia was investigated for a possible immunodeficiency. Lymphocyte

subpopulations and intraand extracellular HLA-I expression were analyzed by flow

cytometry. Subcellular distribution of HLA-I was examined by confocal microscopy. An

HTLV-1 transformed T lymphocyte cell line was generated and used in some of these

analyses. Western blotting and Sanger sequencing was applied to establish the final

diagnosis.

Results: Initial laboratory test showed severe T cell lymphocytopenia of CD8+ cells with

very low naïve and central memory CD4+ cells. B and NK cells were normal. IgA and IgE

were increased. Proliferation in response to OKT3, PHA or PMA+Ionomycin was severely

impaired. Extracellular, but not intracellular, HLA-I expression levels were reduced.

Confocal microscopy suggested accumulation of HLA-I in Golgi but in later samples HLA-I

expression was normal. Sequencing of WAS detected an unreported mutation coding a

protein predicted to be prone to degradation, as confirmed by Western analysis.

Conclusions: Initial clinical presentation of WAS can sometimes be remarkably similar to

classical combined immunodeficiencies. Platelet count and size should be carefully

assessed to consider WAS. Infection and dexamethasone can reduce HLA-I expression

wrongly leading to the suspicion of bare lymphocyte Type I syndrome.

26 www.sci.cat

Posters


The authors will attend the poster on 16/11/2017: 17:30h; 17/11/2017: 11:00h, 13:30h, 16:30h.

2 Biological immunotherapy and thyroidal disorders

Carla Fernández-Prendes1; Marc Cucurull

2; Sofia España

2; Alejandra Carolina Pérez

3; Aram Boada

4;

Nina Richarz4; Natalia Rivera

4; Olatz Etxaniz

2; Clara Esteve

5; Bibiana Quirant

5; Aina Teniente

5;

Montserrat Marqués3; María Luisa Granada

1; Eva Martínez-Cáceres

1

1Bioquímica Clínica, Hospital Universitario Germans Trias i Pujol, Badalona;

2Oncología Médica, Hospital

Universitario Germans Trias i Pujol, Badalona; 3Endocrinología, Hospital Universitario Germans Trias i Pujol,

Badalona; 4Dermatología, Hospital Universitario Germans Trias i Pujol, Badalona;

5Inmunología, Hospital

Universitario Germans Trias i Pujol, Badalona

The increasing use of antibodies targeting immune check-points inhibitors (CPI) especially

antiprogrammed cell death-1 (anti-PD1), anti programmed death-ligand 1 (anti-PDL1) or

anti-cytotoxic Tlymphocyte– associated antigen 4 (anti- CTLA4) in the treatment of

malignancies like lung cancer or melanoma has led to a parallel increase on immune-

related adverse effects such as thyroidal disorders, many of them autoimmune.

The aim of this study was to analyze the incidence of thyroidal alterations in patients

treated with CPI in a tertiary hospital of the Barcelonès-Nord.

Method: retrospective analysis of patients treated with CPI between 2014 and 2017 The

prevalence of thyroidal alterations was evaluated using thyrotropin rr[0,35-4,94] and free

thyroxine (fT4 rr[0,7-1,48ng/dl]). 290 patients were divided according to the drug

received.Thyroidal disorders were qualified as important when TSH (μUI/ml) was <0,1 or

>10 and /or fT4 (ng/dl) < 0,5 or >2.

Results: 63,8% patients suffered lung cancer, 18,3% melanoma, 6,9% urothelial

carcinoma and 11% other types of cancer. The table shows patients grouped according to

the drug received, its mechanism of action and the severity of thyroidal alterations. Drug Mechanism of actión Nº of patients Severity Thyroidal disorders (%)

PEMBROLIZUMAB anti-PD1 43 Mild: 7 Important: 10 39.53

NIVOLUMAB anti-PD1 139 Mild: 25 Important: 25 35.97

ATEZOLIZUMAB anti-PDL1 36 Mild: 8 Important: 7 41.66

DURVALUMAB anti–PDL1 11 Mild: 1 Important: 0 9.09

IPILIMUMAB anti-CTLA4 5 Mild: 0 Important: 1 20.00

IPILIMUMAB/NIVOLUMAB anti-CTLA4; anti-PD1 8 Mild: 2 Important: 1 27.27

Others 48

The prevalence of thyroid disorders varied widely depending on the drug received (from

41,66% in patients treated with Atezolizumab to 9,09% in patients treated with

Durvalumab). Thyroidal disorders were not associated to the mechanism of action of the

drug received.

Conclusion: Given the high frequency of thyroidal pathology in patients treated with CPI,

it is important to protocol the monitoring of thyroidal hormones and anti-thyroidal

antibodies periodically.

www.sci.cat 27

Posters


3 Immunological characterization of IgA nephropathy patients

Clara Esteve Cols1,2

; Fredzzia Amanda Graterol Torres3; Aina Teniente Serra

1,2; Bibiana Quirant

Sánchez1,2

; Jordi Ara del Rey3; Eva Mª Martínez-Cáceres

1,2

1Immunology Department Hospital Universitari Germans Trias i Pujol, Badalona (Spain);

2Department of Cell

Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Bellate; 3Nephrology Department

Hospital Universitari Germans Trias i Pujol, Badalona (Spain)

IgA Nephropathy (IgAN) is the leading form of primary glomerulonephritis affecting

glomerular mesangium, which cause proteinuria, hematuria, hypertension and reduction of

renal glomerular filtrate. Approximately 40% of cases are related to end-stage renal failure,

requiring either dialysis or renal transplantation. Although the pathogenic mechanisms

responsible of the disease are unknown, it has been postulated that the immune system

plays an important role. The gold-standard technique for IgAN diagnosis is renal biopsy. In

the last years, the better knowledge of the disease, has led several authors to describe

serum biomarkers that may be useful for diagnosis and prognosis, such as levels of

partially degalactosilated IgA1 (Gd-IgA1), of specific IgG against Gd-IgA1, as well as

sCD89 and Gd-IgA1 immune complexes. Until now, it has not been performed an

exhaustive analysis of peripheral leukocyte subpopulations and CD89 expression on

monocytes, which are the objectives of this study.

A prospective study of 22 patients diagnosed of IgAN by renal biopsy has been performed.

By flow cytometry of whole blood, immunophenotype of leukocyte subpopulations and

CD89 expression in three monocytes subpopulations has been characterized. Analysis of

serum levels of Gd-IgA1 has been performed with commercial kit of ELISA, Gd-IgA1

Assay kit-IBL.

The results of this study have shown that those patients with poor renal function and more

severe renal biopsy have lower Mean Fluorescence Intensity (MFI) of CD89 on non-

classical monocytes. The immunophenotype showed that patients had a higher

percentage of activated and efector memory CD4+ and CD8+ lymphocytes, lower

percentages of B transitional lymphocytes and plasmablasts, and higher percentages of

NK lymphocytes CD56dimCD16+ and myeloid dendritic cells.

In conclusion, this preliminary study shows that MFI of CD89 on non-classical monocytes

could be used as a prognostic biomarker of IgAN. In parallel, the immunophenotype

maybe useful for IgAN diagnosis.

28 www.sci.cat

Posters


4 New insights into Cyclin D3 interactions in the development of

Type 1 Diabetes

Celia Vived1; Celeste Santos

1; MªAngeles de la Torre

1; Leire Egía-Mendikute

1; Estela Rosell-Mases

1;

Joan Verdaguer1; Conchi Mora

1

1IRB Lleida/Universidad de Lleida

Type 1 diabetes (T1D) is an autoimmune condition caused by the lymphocyte-mediated

destruction of the insulin-producing β cells in pancreatic islets. Our group has previously

described that cyclin D3 is downregulated in a dose-dependent manner in β cells by

leukocyte infiltration into the islets of the nonobese diabetic (NOD) type 1 diabetes-prone

mouse model. This protein, classically related to cell proliferation, is targeted in

autoimmune diabetes and exerts a protective role on β cells by promoting their survival

without affecting proliferation.

We have identified by the yeast two-hybrid technology (Y2H) a number of molecules other

than the CDKs that physically interact with cyclin D3. We have seen that cyclin D3

interacts with proteins involved in several physiological processes such as the UPR

(unfolded protein response) which promotes the production of reactive oxygen species

(ROS) in the beta cell, causing oxidative stress and damaging the mitochondrial function.

Other processes in which these candidates are involved are the mitochondrial function and

the redox balance and, vesicle trafficking. Cyclin D3-mediated regulation of insulin

secretion through vesicle trafficking would account on the relevance of cyclin D3 in islet

physiology.

We are interested in the molecules obtained from Y2H that are not involved in the cell

cycle, in order to dissect metabolism and viability and cell cycle. In this way, we can

analyse this new role of cyclin D3 in the viability and fitness of beta-pancreatic cells and it

helps the translation into future therapeutic targets for T1D.

www.sci.cat 29

Posters


5 Sustained Spontaneous Partial Remission in a Pediatric

Patient with Type 1 Diabetes

Silvia Rodríguez-Fernández

1; Marta Murillo

2; Mireia Fonolleda

1; Adrian Villalba

1; Federico Vázquez

3;

Joan Bel2; Marta Vives-Pi

1,4

1Immunology Division, Germans Trias i Pujol Research Institute, UAB, Badalona;

2Pediatrics Section,

Germans Trias I Pujol University Hospital, Badalona; 3Endocrinology and Nutrition Section, Germans Trias i

Pujol University Hospital, Badalona; 4CIBERDEM, Barcelona

Type 1 diabetes (T1D) is a metabolic disorder of unknown etiology that results from

autoimmune beta-cell destruction. Shortly after initiation of insulin therapy, patients may

experience a spontaneous partial remission phase called honeymoon, in which insulin

requirements decrease and glycemic control improves during a few months. It is

hypothesized that honeymoon occurs after a transient recovery of immune tolerance,

which allows for beta-cell regeneration.

We present a 18-year-old girl, diagnosed at 12 years with T1D, with spontaneous partial

remission sustained for more than 6 years. At diagnosis, glycemia was 487 mg/dl without

ketoacidosis, HbA1c levels were 13.5% and plasma C-peptide (a molecule secreted

alongside insulin which serves to monitor residual betacell function) was 1 ng/ml.

Autoantibodies to glutamic acid decarboxylase and islet antigen 2 were negative. HLA

typing was DRB1*04:01 (DR4) and DQB1*03:02 (DQ8). She was transitioned to basal-

bolus subcutaneous insulin regimen. One month after diagnosis, HbA1c was 8%,

improving to 5.2% afterwards. During the next 6 years, HbA1c was <5.7% and insulin

requirements were ≤0.8 UI/kg/24h, and stimulated Cpeptide was 1.3 ng/ml from a basal 1

ng/ml level. Autoantibodies remained negative, except for autoantibodies to insulin.

Lymphocyte subsets analysis showed reduced T-lymphocyte counts, normal CD4/CD8

ratio and decreased percentages of Treg, both memory and activated. Betatrophin level,

an angiopoietin-like family hormone increased in T1D, was significantly augmented 6

years after diagnosis compared to controls. Monogenic diabetes was ruled out by genetic

study of glucokinase, HNF1A and HNF4A genes, the most frequent types of diabetes

maturity-onset diabetes of the young (MODY). The patient is currently in partial remission,

requiring an insulin dosage of 0.38 UI/kg/day and HbA1c 5.2%. Therefore, a balance

between low-grade autoimmunity and regeneration seems to be maintained in

honeymoon. The assessment of the patient progress will help elucidate the immunological

significance of this stage.

30 www.sci.cat

Posters


6 A new case of IgE multiple myeloma

Sergio Navarro Velázquez1; Virgínia Mas Bosch

1; Francisco Morandeira Rego

1; Elisabet Poyatos

Canton1; Sonia Barrachina Moles

1; Cristina Fuertes Purificacion

2; Jordi Bas Minguet

1

1Department of Immunology, Bellvitge University Hospital;

2Department of Clinical Biochemistry and

Immunology, Bellvitge University Hospital

Introduction: Multiple myeloma (MM) is a neoplastic syndrome characterized by

proliferation of plasma cells in the bone marrow and the subsequent appearance of a

monoclonal immunolgobulinin the patient's serum. The four typical clinical manifestations

of MM include: hypercalcaemia, renal insufficiency, anaemia and osteolytic lesions.

Different types of MM can be distinguished: IgG and IgA are the most frequent (55% and

20%), followed by light chain-secreting MM (18%). The non-secretory, the IgD and the IgM

type are much rarer (4.4%, 1.7% and 0.3%), and IgE MM is extremely uncommon (<

0.1%). Recently, we observed a band corresponding to IgE kappa while analyzing the

serum of a 74 year-old man with an intracranial hematoma, lumbar protrusion and a

history of stroke.

Objective: To verify the IgE kappa isotype of the band, as it is extremely rare.

Materials and methods: Capillary electrophoresis and immunotyping (IT) were carried

out with Capillarys 2 flex piercing, Sebia. IFIX with specific IgD and IgE antisera was

performed with Hydrasis 2, Sebia. Total IgE quantification was carried out with

ImmunoCAP® (Thermo Fisher Scientific).

Results: Capillary electrophoresis showed a monoclonal component (MC) < 3.5 g/L in

the gamma region. To establish the isotype of the peak we performed IT and we observed

a peak decrease in kappa, but not in G, A or M. A subsequent IFIX detected a clear band

of IgE kappa isotype of 1.227.200 karb.u./L.

Conclusions: We verified the presence of IgE kappa in the patient’s serum, based on

these findings. We suggest a role for MM in the development of the patient’s clinical

picture that needs a more detailed study.

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Posters



7 Comparison of anti-PLA2R autoantibodies detected by ELISA

and indirect immunofluorescence

Elisabet Poyatos

1; Francisco Morandeira

1; Juliana Bordignon

2; Xavier Fulladosa

2; Sergio Navarro

1;

Virgínia Mas1; María Jesús García

1; Julia Saiz

1; Jordi Bas

1

1Immunology Department. Hospital Universitari de Bellvitge;

2Nephrology Department. Hospital Universitari

de Bellvitge

Introduction: The detection of anti-phospholipase A2 receptor antibodies (PLA2R)

identifies a subset of patients with primary membranous glomerulonephritis (MGN) who

can benefit of immunosuppressive treatment and allows therapy monitoring. ELISA and

immunofluorescence (IIF) are currently available techniques but there are few data

comparing its performance.

Objectives: To compare anti-PLA2R detection by both techniques and to validate the

manufacturer's recommended ELISA reference limits in our clinical setting.

Material and methods: 83 samples from 71 patients were analyzed in parallel by ELISA

and IIF on PLA2R-transfected cells (Euroimmun). The relationship between ELISA and IIF

was established using a Cohen kappa index with Analyze-it® software. 33 samples from

healthy volunteers were also included. A clinical data review was also performed.

Results: Seventy-six samples (91.6%) from 64 patients obtained a concordant

classification by the two methods. Discordant results were obtained in seven (0.084%)

(kappa index: 0.79), 3 (IIF-positive) of which yielded significant ELISA values albeit they

didn’t reach the manufacturer's reference limit. 31 healthy individuals showed undetectable

ELISA values. All patients with concordant results and PLA2R antibodies had MGN

(n=15). The remaining 49 patients were PLA2R-negative (59% MGN -59% remission-,

37% no MGN, 4% no information). The seven discordant results comprised three patients

in remission of MGN (IIF+, ELISA-), two in relapse (one IIF doubtful, ELISA+; one IIF+,

ELISA-), one in partial remission (IIF+, ELISA-) and one in doubtful remission (IIF+, doubtful

ELISA).

Conclusions: The two techniques showed an overall good agreement. Anti-PLA2R

antibody tests had a high positive predictive value. The PLA2R ELISA was slightly less

sensitive than IIF but it may be more objective and convenient for therapy monitoring. In

the few discordant cases, the clinical features showed a better relationship with ELISA.

The reference limits established by the manufacturer may be lowered. A larger study is

needed to confirm these points.

32 www.sci.cat

Posters


8 Optimization of cytogram interpretation for HLA-B*27 testing

by flow cytometry compared to PCR genotyping

Elisabet Poyatos1; Ariadna Padró

2; Sònia Barrachina

1; Rafel Mayol

1; Virgínia Mas

1; Francisco

Morandeira1; Sergio Navarro

1; Jordi Bas

1

1Immunology Department. Hospital Universitari de Bellvitge;

2Genetics Department. Hospital Universitari de

Bellvitge

Introduction: HLA-B27 allele is present in more than 90% in patients with ankylosing

spondylitis (AS) and less than 8% in European population. Since flow cytometry (FCM)

may give false positive results due to cross reaction with other surface antigens (B7, B40)

the confirmation of positives by allele-specific PCR melting assay is the most convenient

procedure for HLA B27 testing. Cut-off setting is critical to discriminate true negative

samples by FCM in that further genotyping is not necessary. This allow to reduce the

number of genetic tests, thus optimizing economical resources.

Objectives: Optimization of FCM minimum percentage of cells above cut-off values,

according to DNA genotyping, with assessment of patient clinical features.

Materials and methods: Data from 180 patients tested for HLA-B27 by CD3-PE/anti-HLA-

B27-FITC incubation and FCM (FACScalibur, Becton Dickinson) plus real time PCR with

melting curve analysis (ECO, Illumina) from 2015 to 2016 were included. Percentage of

cells above cut-off (set at 70 green fluorescence units) and the mean immunofluorescence

channel (MIF) were recorded in samples with (n=130) or without (n=50) HLAB27

genotype. Clinical assessment was performed in these patients and in a further group of

50 patients without available cytograms.

Results: All HLA-B27 negative samples by PCR genotyping showed less than 68% cells

over cut-off (except three cross-reacting) and a MIF less than 100 units (30% between 70

and 100) by FC. Mean percentage of positives was 94% ± 4.51SD, and a MIF>70. All AS

patients were HLA-B27 (27% of positive samples). Other spondylopathies accounted for

21% of positives and 13% of negatives.

Conclusions: Lowering the percentage of cells over cut-off to 67% accommodated 100%

of positives and prevented DNA testing of 47 samples. MIF values are not useful to

discriminate positive samples. A prospective study is ongoing to validate this cut-off.

www.sci.cat 33

Posters



9 Development of a CCR5-Δ32/Δ32 inventory within the best cell

quality umbilical cord blood units of the Spanish Registry

Emma Enrich1; Francisco Vidal

1; Francesc Rudilla

1; Jose Luis Caro

1; Maria Piron

1; Nina Borràs

1;

Laura Mongay1; Sílvia Sauleda

1; Sergi Querol

1

1Banc de Sang i Teixits, Vall d’Hebron Institut de Recerca, Universitat Autònoma Barcelona (VHIR-UAB)

A homozygous 32-base pair deletion in the gene that codifies for the chemokine receptor

CCR5 confers natural resistance to CCR5-tropic HIV-1 infection. The case of a patient with

HIV infection and haematological disease, who underwent hematopoietic transplantation

from a CCR5-Δ32/Δ32 unrelated donor and who has not shown evidence of HIV-1

infection after the procedure, has suggested the possibility to use CCR5-Δ32/Δ32 umbilical

cord blood in HIV patients in need of hematopoietic transplantation. In this regard, the

main objective of this study was to achieve an inventory of CCR5-Δ32/Δ32 cord blood

units from the Spanish Registry and to study their biological characteristics of interest.

CCR5 variants of the 20,236 best cell quality units of the Spanish Registry were

characterized using a twosteps strategy, based on two in-house developed tests: Real-

Time PCR for the screening of all units and then capillary electrophoresis to confirm

sequence of those detected as positive for the CCR5-Δ32 variant. In addition, data

analysis was performed to determine CD34+ and total nucleated cells counting, donors

geographical and ancestral origin and also blood type, gender and HLA typing properties

of the units according their CCR5 genotype.

The results showed 130 (0.64%) units homozygous for the deletion, 2,646 (13.08%) were

found to be heterozygous and 17,460 (86.28%) did not present the mutation. A significant

association was found among donor's ancestral origin and the mutation, with a higher

percentage of CCR5-Δ32 units with European ancestry. Interestingly, statistically

significant lower amount of CD34+ cells was found in the CCR5-Δ32 homozygous units.

In summary, we have obtained 130 CCR5-Δ32/Δ32 cord blood units in an inventory of

20,236 ready for use in transplantation. Further studies are required to determine the

clinical impact of lower amounts of CD34+ cells found in these units.

34 www.sci.cat

Posters



10 Immunomodulatory effects of an early life prebiotic

supplementation to suckling rats

Azagra-Boronat I.1,2

; Massot-Cladera M.1,2

; Knipping K.3; Tims S.

3; van’t Land B.

3,4; Stahl B.

3; Knol J.

3;

Garssen J.3,5

; Franch A.1,2

; Castell M.1,2

; Rodríguez-Lagunas M.J.1,2

; Pérez-Cano F.J.1,2

1Departament de Bioquímica i Fisiologia. Facultat de Farmàcia i Ciències de l’Alimentació. UB.;

2Institut de

Recerca en Nutrició i Seguretat Alimentària (INSA-UB), Santa Coloma de Gramenet, Spain; 3Nutricia

Research, Utrecht, the Netherlands; 4University Medical Centre Utrecht/Wilhelmina Children's Hospital,

Department of Pediatric Immunology, Utrecht, the Netherlands; 5Utrecht Institute for Pharmaceutical

Sciences, Faculty of Science, Utrecht University, Utrecht, the Netherlands

Early life immune and microbiota development are influenced by several bioactive and

prebiotic factors present in breast milk. Aiming to mimic breast milk composition, infant

formulas include dietary ingredients with this prebiotic effect. The aim of this work was to

study the influence of a supplementation with an Early Life Prebiotic (ELP) during the

suckling period on the development of immunity and the microbial composition and

functionality.

In order to achieve this objective, neonatal Lewis rats received either ELP or PBS (control)

through oral gavage during the period of strictly suckling (until day 16 of life). The levels of

the different Ig classes (IgG, IgM, IgA) and subclasses (IgG1, IgG2a, IgG2b and IgG2c)

were quantified using a ProcartaPlex® Multiplex Assay. Microbiota composition was

evaluated by sequencing the rRNA V3-V4 from the 16S RNA in caecal samples. The urine

metabolomic analysis was performed by NMR and the short chain fatty acids (SCFA)

quantification in caecal samples by GC-MS.

In suckling rats increased concentrations of plasmatic Th1-associated Ig (IgG2b+IgG2c in

rat) was detected in rats receiving ELP as compared to control without affecting Th2-

associated Ig (IgG1+IgG2a in rat) at both day 8 and 16 of life. In addition, significantly

(p<0.05) increased plasmatic IgA levels were detected in rats receiving ELP as compared

to control rats. Within ELP receiving rats changes in some microbial groups were detected,

for instance, it boosted the proportion and diversity of Lactobacillus species. Moreover, it

increased the relative proportion of butyric acid, but not the other SCFA. The urinary

metabolome was also clearly affected in those animals due to ELP, affecting the

proportion of several microbial and endogenous metabolites.

In conclusion, ELP modulates both immunological as well as microbial development of

suckling rats, suggesting importance of this type of compounds in early life.

www.sci.cat 35

Posters

Innate Response 9 - 16 The authors will attend the poster on 16/11/2017: 17:30h; 17/11/2017: 11:00h, 13:30h, 16:30h.

11 Resistin in ascitic fluid of patients with cirrhosis modifies

innate immune function

Lídia Perea1; Juan Camilo Nieto

1; Germán Soriano

2; Elisabet Sánchez

2; Eva Román

2; Carlos

Guarner2; Carlos Zamora

1; Elisabet Cantó

1; Cándido Juárez

1; Silvia Vidal

1

1Department of Immunology, Institut de Recerca and Hospital Santa Creu i Sant Pau, Barcelona;

2Department of Gastroenterology, Hospital Santa Creu i Sant Pau, Barcelona

Introduction: Patients with decompensated cirrhosis have complications such as

spontaneous bacterial peritonitis (SBP) that lead to a defective immune response. We

hypothesized that this defect could be induced by the content of the ascitic fluid.

Methods: We tested the effect of cell-free sterile ascites (SA) and SBP at diagnosis and

after treatment on healthy neutrophils (oxidative burst and NETosis). We measured

resistin, IL-6 and IL-1Ra levels in ascitic fluids and we correlated them with clinical

characteristics. We also tested the effect of priming with resistin on healthy neutrophils and

monocytes.

Results: We observed that healthy neutrophils cultured with SBP ascitic fluid exhibited

lower oxidative burst levels and lower NET production that healthy neutrophils cultured

with SA. NET production was significantly increased in patients treated with antibiotic. At

diagnosis, SBP contained higher resistin, IL-6 and IL-1Ra levels than SA. After antibiotic

treatment, resistin and IL-6 levels tended to decrease without reaching SA levels. We

found negative correlations between resistin levels and neutrophil function. Furthermore,

resistin was associated with clinical characteristics of SBP and severity of cirrhosis.

Interestingly, we demonstrated that monocytes pre-treated with resistin and exposed to

E.coli expressed lower percentage of CD14+CD16+ cells and TNF-α production compared

to untreated monocytes exposed to E.coli. Finally, we also associated the high resistin and

IL-6 levels in SBP at diagnosis with the poor prognosis of SBP episode.

Conclusions: The content of ascitic fluids from patients with SBP reduces the capacity of

healthy neutrophils. Resistin could be one of the major players in this reduction.

36 www.sci.cat

Posters


12 Altered phenotypic and functional features of healthy donor

monocytes with bound platelets

Mariscal-Rodríguez A.1; Zamora C.

.1; Martínez-Martínez L.

1; Perea L.

1; Ortiz M.A.

1; Cantó E.

1; Vidal S.

1

1Hospital de la Santa Creu i Sant Pau

There are aggregates of platelets with lymphocytes and monocytes in non-pathological

conditions. We have previously shown that the frequencies of these aggregates are

altered in inflammatory chronic diseases. These alterations can have pathological

consequences, because current findings suggest that the binding of platelets regulates the

function of lymphocytes and monocytes.

In this research project we have analysed the possible differences between monocytes

with or without bound platelets from healthy donors at the level of the expression of

surface markers, the secretion of anti and proinflammatory cytokines (IL-10 and TNF-α)

and the phagocytic ability by flow cytometry.

We found that monocytes with or without bound platelets had a different expression of

CD11c and CD54, as adhesion markers, and HLA-DR, as a molecule involved in antigen

presentation. We also found that monocytes with bound platelets secreted more IL-10 than

monocytes without bound platelets. Besides, those monocytes with activated platelets

secreted even more IL-10. However, TNF-α secretion was comparable in monocytes with

or without bound platelets. Finally, we observed that monocytes with bound platelets

phagocyted E. coli cells more efficiently than monocytes without bound platelets.

Our findings suggest that monocytes with bound platelets had a modified adhesion

capacity, antigen presentation ability, cytokine secretion and phagocytosis capacity. Our

currently global analysis of the function suggests that monocytes with bound platelets have

an anti-inflammatory role. Further experiments will be performed to determine whether the

platelet binding modifies monocyte function or whether the platelet binds those monocytes

with an altered phenotype.

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Posters



13 Gene expression changes in intestinal epithelial barrier

molecules after leptin or adiponectin supplementation in

suckling rats

Abril-Gil M.1,2

; Grases-Pintó B.1,2

; Marín-Morote L.1,2

; Castell M.1,2

; Rodríguez-Lagunas M.J.1,2

; Pérez-

Cano F.J.1,2

; Franch A.1,2

1Departament de Bioquímica i Fisiologia, Facultat de Farmàcia i Ciències de l’Alimentació (UB);

2Institut de

Recerca en Nutrició i Seguretat Alimentària (INSA·UB)

The gastrointestinal tract not only has a central role in nutrients digestion and absorption

but it is also a physiological key barrier between the body and the outer environment. This

physical, microbiological and immune barrier involved in such protection is in continuous

development during early life. Breastfeeding plays a key role in the maturation of the

intestinal barrier and milk adipokines may influence in this process.

The present study aimed to evaluate the adipokines’ role in the maturation of the intestinal

barrier in rats by shedding light on their influence on the innate immunity development and

the presence of the intestinal epithelial barrier proteins.

For this purpose, suckling Wistar rats were daily supplemented with leptin, adiponectin or

vehicle throughout the suckling period. In the middle (day 10) and at the end (day 21) of

suckling, a fragment of small intestine was obtained to determine the intestinal expression

of genes related to the innate immune defence (mucin-2 and mucin-3) and to the epithelial

barrier integrity (ZO-1, occludin, claudin-2 and claudin-4) by RT-PCR. At the end of

suckling, the gene expression of toll-like receptors 2 and 4 was also evaluated.

The results revealed a significant increase in the intestinal gene expression of claudin-4

and mucin-2 due to the leptin supplementation only during the first half of the suckling. On

the contrary, even though adiponectin supplementation did not modify the gene expression

of the studied molecules in early suckling period, it was able to enhance the innate

defence just after weaning by increasing mucin-2 and mucin-3 gene expression in

comparison with the reference group (p<0.05). The TLR gene expression was not modified

by adipokines supplementation.

From these results, we can conclude that the leptin or adiponectin supplementation may

play a role in the maturation of the intestinal epithelial barrier in suckling rats.

38 www.sci.cat

Posters



14 Expression analyses of porcine Tumor Necrosis Factor

receptor 2 variants.

Sebastián G. Kuguel1; Mireia Uribe-Herranz

1; Cristina Costa

1

1Institut d’Investigació Biomèdica de Bellvitge (IDIBELL), L’Hospitalet de Llobregat, Barcelona

TNF participates in transplant rejection by promoting immune cell activation and

inflammation. To elucidate its mechanisms in xenotransplantation, we previously cloned

the cDNA of porcine TNF-Receptor 2 (pTNFR2) and found four isoforms generated by two

alternative splicings. The processing of ΔE7-10 splicing resulted in loss of the

transmembrane region and production of soluble variants, whereas the ΔE4 splicing

shortened the TNF-binding domain compromising this function.

We hypothesized that the various isoforms display distinct activities and performed

expression studies to elucidate their roles.

First, we carried out quantitative RT-PCR to determine the relative amount of the two

alternative splicings in porcine aortic endothelial cells (PAEC) and porcine costal

chondrocytes (PCC) in resting and cytokine-stimulated conditions. The predominant

isoform was the full receptor, but pro-inflammatory IL-1α and TNFα upregulated the mRNA

expression of all isoforms. For subcellular localization studies, a series of experiments

based on immunofluorescence and confocal microscopy were conducted with porcine cells

and cells genetically-modified to express the membrane-bound isoforms. Our polyclonal

anti-pTNFR2 antibodies recognized both pTNFR2 and pTNFR2ΔE4 by immunostaining.

Co-staining with phalloidin in PAEC confirmed that only a small proportion of the receptor

was expressed on the cell surface (14% average), whereas recombinant overexpression

of pTNFR2 substantially increased the co-localization at the plasma membrane (40%).

Transient transfection in Hela cells of pTNFR2-EYFP and pTNFR2ΔE4-EYFP fusion

proteins containing the TNF-binding and transmembrane regions produced a very different

pattern. A high proportion of the pTNFR2-EYFP co-localized with phalloidin at the plasma

membrane (~37%), whereas all pTNFR2ΔE4-EYFP remained inside the cell. In fact, the

expression pattern of pTNFR2ΔE4, but not of pTNFR2, was compatible with a

predominant expression in the endoplasmic reticulum.

Thus, we reveal the expression pattern of pTNFR2 isoforms as it may have an impact on

the process of xenograft rejection.

www.sci.cat 39

Posters


15 Validation and characterization of a genetic biomarker for a

vitamin D3-induced tolerogenic dendritic cell-based therapy in

multiple sclerosis

Juan Navarro-Barriuso1,2

; María José Mansilla1,2

; Chiara Bernardi1,2

; Bibiana Quirant-Sánchez1,2

; Aina

Teniente-Serra1,2

; Silvia Presas-Rodríguez3,4

; Cristina Ramo-Tello3; Eva María Martínez-Cáceres

1,2

1Immunology Division, Germans Trias i Pujol University Hospital and Research Institute.;

2Department of

Cellular Biology, Physiology and Immunology, Universitat Autònoma de Barcelona.; 3Multiple Sclerosis Unit,

Department of Neurosciences, Hospital Universitari Germans Trias i Pujol.; 4Department of Medicine.

Universitat Autònoma de Barcelona

Background: A breach of tolerance against determined self-peptides may lead to a

complex and pathologic disorder of the immune system, causing autoimmune diseases

such as multiple sclerosis (MS). Novel autologous vitamin D3-induced tolerogenic dendritic

cell (vitD3-tolDC)-based therapies have become promising therapeutic alternatives to

conventional treatments for autoimmune diseases by their potential ability to restore

tolerance against the peptide they present. To guarantee the safety and tolerogenicity of

vitD3-tolDC prior to administration into patients in a clinical trial, it is important to have fast,

reliable and robust biomarkers. In this context, the so-named Gene 2, encoding an

structural protein, could be a good candidate.

Objective: To validate and characterize Gene 2 as a potential biomarker of vitD3-tolDC.

Methods: Twenty four monocyte-derived dendritic cell differentiations of immature (iDC),

mature (mDC) and vitD3-tolDC from healthy donors and 10 from MS patients were

generated and characterized phenotypically and functionally (inhibition of allogeneic

PBMC proliferation). RNA was extracted and retrotranscribed into cDNA, and the

expression of Gene 2 was analyzed by qPCR. Expression was considered differential

when logFC >0.5. The expression and distribution of the protein encoded by Gene 2 was

characterized by immunocytochemistry (ICC) in 4 differentiations from healthy donors.

Results: In HD, Gene 2 was overexpressed in vitD3-tolDC compared to mDC (logFCHD

1.02 ± 0.22), while the mean expression between iDC and mDC remained unchanged

(logFC <0.5). The same behavior was observed in patients (logFCpatients 0.51 ± 0.30). At

the protein level, Gene 2 presented a 3-fold expression in vitD3-tolDC respect iDC and

mDC. Statistical significance was reached in all cases (p<0.05).

Conclusions: The results suggest that Gene 2 may be a robust biomarker to characterize

vitD3-tolDC products. Nevertheless, its characterization should be performed in samples

from patients included in a clinical trial in order to confirm these results.

40 www.sci.cat

Posters


16 Blimp-1 and neonatal-Fc-receptor gene expression as

biomarkers to study the intestinal barrier maturation in

suckling rats

Massot-Cladera M.1,2

; Abril-Gil M.1,2

; Grases-Pintó B.1,2

; Azagra-Boronat I.1,2

; Rodríguez-Lagunas

M.J.1,2

; Castell M.1,2

; Franch A.1,2


1Departament de Bioquímica i Fisiologia, Facultat de Farmàcia i Ciències de l’Alimentació, UB;

2Institut de

Recerca en Nutrició i Seguretat Alimentària (INSA-UB), Universitat de Barcelona

The immature gastrointestinal tract from neonatal mammals is adapted to the digestion of

milk and is permeable to essential milk-borne bioactive macromolecules present in

mother’s milk such as IgG antibodies. It is well known that the neonatal-Fc-receptor (FcRn)

mediates the binding and transfer of IgG across the small intestine during the

breastfeeding period. Moreover, the transcription factor B-lymphocyteinduced maturation-

protein-1 (Blimp-1) has been recently suggested that is also involved in the intestinal

maturation in rats.

In this sense, the aim of the present study was to establish the process of maturation of

the intestinal barrier in rats by means of monitoring the expression of both Blimp-1 and

FcRn genes along the suckling period.

In order to achieve this purpose, a 5 mm fragment of the middle part of the small intestine

was collected from strictly suckling-animals (at days 8, 10 and 14 of life) and weaned (at

day 21) rats. Gene expression of both molecules was evaluated by Real Time-PCR using

TaqmanÒ specific probes and primers.

FcRn expression in the small intestine during suckling was similar in the first two weeks of

life, but when the ingestion of solid food diet started, its levels were much lower (p<0.05

vs. day 14). The intestinal Blimp-1 expression showed an increasing pattern during the first

ten days of suckling (p<0.05), but its expression was suddenly lowered later (day 14) and

kept constant until the end of the studied period (p<0.05 vs. day 10).

From these results it can be concluded that the maturation of the small intestine in rats can

be defined by the different expression of FcRn and Blimp-1 genes, which show a

characteristic pattern along the suckling period. The modulation of these biomarkers can

help to evaluate the influence of compounds on the intestinal maturation process.

www.sci.cat 41

Posters



17 Modulatory role of platelets in lymphocyte populations of

psoriasis patients

Maria Teresa Sanz Martínez1; Carlos Zamora Atenza

1; Emma Mata

1; Miguel Ángel Sánchez Martínez

1;

Luis Puig Sanz1; Cándido Juárez Rubio

1; Silvia Vidal Alcorisa

1; Esther Moga Naranjo

1

1Hospital de la Santa Creu i Sant Pau

Psoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated

with Th17 cells. Th17 cells produce IL-17A among other cytokines, which seems to have a

prominent pathogenic role in skin inflammation.

Platelets are closely linked to the pathogenesis of inflammatory skin diseases, the degree

of platelet activation correlates with disease activity. It has been recently proposed that

platelets can modulate cytokine production by binding to lymphocytes in other

inflammatory diseases such as rheumatoid arthritis. Our working hypothesis is that

platelets could play an important role in the modulation of immune cells of psoriasis

patients.

For this purpose we analyzed the percentage of lymphocytes with bound platelets through

the expression of CD41a (a specific marker of platelets) on PBMCs from control volunteers

(HC) and psoriasis patients (Pso) by flow cytometry. In order to compare the percentages

of platelet-linked lymphocytes populations, we firstly investigated the total percentage of

lymphocyte subpopulations. A significant higher percentage of CD19+ lymphocytes was

observed in Pso, however the percentage of CD8+ and CD4+ lymphocytes were similar in

Pso and HC. Subsequently, we compared the phenotype of CD4+ lymphocytes. We next

analyzed CD161 expression, a marker described in cells with potencial capacity to

produce IL17. Our results showed a significant higher percentage of CD4+CD161+ cells in

Pso than in HC. Finally, we analyzed the percentage of lymphocytes with bound platelets.

Our results showed that Pso had a higher percentage of CD8+CD41a+ lymphocytes and

lower percentage of CD4+CD41a+ lymphocytes than HC. We did not observe diferences

in the percentages of CD19+CD41a+ lymphocytes. Interestingly, the lower percentage of

CD4+CD41a+ lymphocytes in Pso was due to a decrease in the percentage of

CD4+CD41a+CD161+ lymphocytes.

The higher percentage of CD4+CD161+ lymphocytes with a lower linkage to the binding of

platelets, could explain the increase of IL17 production reported in psoriasis patients.

42 www.sci.cat

Posters



18 The supplementation of TGF-β2, EGF or FGF21 in suckling rats

influences mesenteric lymph node cell composition

Paulina Torres-Castro1,2

; Blanca Grases-Pintó1,2

; Mar Abril-Gil1,2

; Margarida Castell1,2

; Francisco J.

Pérez-Cano1,2

; Àngels Franch1,2

1Departament de Bioquímica i Fisiologia, Facultat de Farmàcia i Cièncias de l'Alimentació, UB;

2Institut de

Recerca en Nutrició i Seguretat Alimentària (INSA-UB), Universitat de Barcelona (UB)

The immune system starts its development during pregnancy and keeps the maturation in

the postnatal period. Breast milk provides bioactive components, such as specific growth

factors, that promote the maturation of the immune system in the newborn.

The aim of the present study was to ascertain whether supplementation with transforming

growth factor (TGF)-β2, epidermal growth factor (EGF) or fibroblast growth factor 21

(FGF21), components present in breast milk, to suckling rats could influence the

lymphocyte composition of mesenteric lymph nodes (MLN). For this purpose, suckling

Wistar rats were daily supplemented by oral gavage with TGF-β2, EGF, FGF21 or vehicle

(reference) throughout the suckling period (three weeks). On days 14 and 21, MLN cell

composition was established by immunofluorescence staining and flow cytometry analysis.

In suckling rats, TCRαβ+ cells constituted the main lymphocyte population in MLN followed

by B cells and, in less proportion, TCRγδ+, NK and NKT cells. Although the proportions of

these lymphocytes were not affected by growth factors supplementation, a decrease in the

percentage of TCRγδ+ cells by EGF was found at day 21 (p<0.05 vs. reference).

Regarding minor lymphocyte subsets, it was on day 14 when changes associated with

supplementation were observed. In particular, the proportion of CD8αβ+ cells was

decreased after supplementation with EGF; whereas TCRγδ+CD8- cell percentage was

increased due to FGF21 intervention.The supplementation during 14 days with any of the

three growth factors studied induced a decrease in the proportion of NK cells expressing

CD8, which reached values similar to those from reference 21-day-old rats.

These results demonstrate that TGF-β2, EGF or FGF21 supplementation in early life is

able to modify the lymphocyte composition in MLN, suggesting their contribution to the

maturation of neonatal intestinal immune system.

www.sci.cat 43

Posters



19 Mild Acid Elution (MAE) vs. Immunoprecipitation (IP): A cell

surface specific approach against the state-of-art method to

elute HLA Class I peptides

Roc Farriol Duran1,2

1Immunologia Cel·lular, Institut de Biotecnologia i Biomedicina, UAB;

2Centre for Proteomics and

Metabolomics, Leiden University Medical Center (The Netherlands)

MHC molecules are virtually expressed on the surface of every cell where MHC-peptide

complexes are the main actors in antigen presentation to T-cells. Hence, the MHC-

peptide-TCR interaction is crucial for the unravelling of the adaptative immune response.

As a set, the presented peptides are named immunopeptidome and they are studied in a

mass spectrometry-based (MS) emerging field called MHC peptidomics. MHC peptide

isolation and purification are crucial for their characterization via MS. Traditionally, the

state-of-art method to isolate MHC peptides has been immunoprecipitation of MHCpeptide

complexes from cell lysates. However, this technique has yield limitations. It requires large

amounts of cells, which apart from making it time consuming and economically restrictive,

are not always available; for instance, in patient samples. Alternatively, there is a method

that has been available since the 90s, Mild Acid Elution (MAE). MAE is based on the

disruption of the β2m subunit from MHC-Class I molecules and the release of the bound

peptides to the supernatant when alive cells are exposed to a citrate shock (pH=3.3). The

simplicity of this procedure makes it much more straightforward and affordable than the

traditional IP’s, that in addition require the use of antibodies. Nevertheless, its cell surface

specificity comes with a greater peptide inespecificity. Seemingly, a greater yield can be

expected thanks to a smaller sample loss and potentially to the iterative capacity of this

approach. The benefits and drawbacks of MAE and IP and their outcome differences are

discussed in this comparative study.

44 www.sci.cat

Posters



20 Spleen lymphocyte composition is modulated by leptin and

adiponectin supplementation in suckling rats

Grases-Pintó B.1,2

; Abril-Gil M.1,2

; Castell M.1,2


; Franch A.1,2

1Departament de Bioquímica i Fisiologia, Facultat de Farmàcia i Ciències de l’Alimentació, UB;

2Institut de

Recerca en Nutrició i Seguretat Alimentària (INSA-UB), Universitat de Barcelona (UB)

Just after birth, the innate and adaptive immune responses are very deficient and is during

the early postnatal period when the immune system develops and acquires its defence

capability against pathogens. It is well known that breast milk is nutritionally adapted to the

requirements of the newborn and plays a role in the immune development process. Among

other bioactive factors present in breast milk, the adipokines, leptin and adiponectin, could

have an effect on the neonatal immune system maturation.

The aim of the present study was to ascertain whether a nutritional supplementation with

leptin or adiponectin in neonatal rats was able to influence the lymphocyte composition of

the spleen during the suckling period.

For this purpose, suckling Wistar rats were daily supplemented by oral gavage for 21 days

with either leptin or adiponectin. During (day 14) and at the end of the suckling period (day

21), spleen lymphocyte composition was established by multiple immunofluorescence

staining and flow cytometry analysis.

During the suckling period, the daily supplementation with leptin or adiponectin influenced

the proportion of the main lymphocyte subpopulations in the splenic tissue, leading to

subset proportions more similar to those of older animals. In particular, at day 14, animals

supplemented with those adipokines had significantly higher proportions of TCRαβ+

lymphocytes and NKT cells than animals receiving vehicle, approaching to composition

found in 21-day-old animals. Moreover, at the end of the suckling, the leptin

supplementation induced a 33% higher proportion of splenic TCRαβ+ lymphocytes with

respect to the percentage found in reference animals at the same age.

Overall, these results suggest that leptin and adiponectin play a role in the maturation of

the immune cells present in the spleen during the suckling period.

www.sci.cat 45

Posters



21 Peripheral lymphocyte subpopulations at onset of type 1

diabetes: imbalance of naïve and memory T cells

Aina Teniente Serra1,2

; Eduarda Pizarro3; Mª Teresa Julián.

3; Marco A Fernández

4; Eva Maria

Martínez-Cáceres1,2

1Immunology Department Hospital Universitari Germans Trias i Pujol, Badalona (Spain);

2Department of Cell

Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Bellaterra; 3Department of

Endocrinology, Hospital de Mataró (Spain); 4Flow Cytometry Facility, Germans Trias i Pujol Research

Institute (IGTP), Campus Can Ruti, Badalona

Introduction: Type 1 diabetes (T1D) is an autoimmune disorder characterized by

destruction of pancreatic beta cells resulting in insulin dependency. Changes in T and B

cell subpopulations in peripheral blood of T1D patients have been described, but a

comprehensive multiparametric flow cytometric analysis is still lacking.

Aim: To identify changes in peripheral blood T- and B- cell compartments in patients at

onset of T1D.

Material and methods: CD4+ and CD8+ T cells (including naïve, central memory, effector

memory and terminally differentiated effector (TEMRA), Th17 and Tregs) and B cells

subsets (naïve, unswitched memory, switched memory and transitional B cells) were

analyzed in peripheral blood of T1D patients at onset (n=26) and healthy donors (HD;

n=40) using multiparametric flow cytometry.

Results: A decrease in the percentage of early and late effector memory CD4+ and CD8+

T cells (T CD4+: p=0.001 and p<0.001, T CD8+: p=0.046 and p<0.001), TEMRA CD4+ and

CD8+ cells (p=0.003 and p=0.004, respectively) was found. In contrast, the percentage of

naïve CD4+ T cells (p=0.010), and percentage and absolute counts of naïve CD8+ T cells

(p<0.001 and p=0.001) were increased in peripheral blood of T1D patients compared with

HD. Moreover, an increase in percentage of total B cells and transitional B cells was

observed in patients compared with HD (p=0.015 and p=0.006, respectively). No changes

were found either in Tregs or in Th17 subpopulations.

Conclusion: The observed changes in the percentage and/or absolute number of

lymphocyte subpopulations support that effector cells migrate to the pancreas participating

in the autoimmune response.

46 www.sci.cat

Posters



22 RORC antagonist inhibits IL-17 production in gut commensal-

specific T cells and mucosa from Crohn’s disease patients

Bassolas-Molina H.1; Raymond E.

2; Labadia M.

2; Wahle J.

2; Harcken C.

2; Hughes R.

3; Turner M.

3;

Smith D..3; Calderón-Gómez E.

1; Esteller M.

1; Carrasco A.

4,5; Esteve M.

4,5; Dotti I.

1; Ferrer-Picón E.

1;

Planell N.1; Masamunt M.C.

1; Gallego M.

1; Barastegui R.

1; Arajol C.

6; Guardiola J.

6; Nabozny G.

2;

Panés J.1; Salas A.

1

1Department of Gastroenterology, IDIBAPS, Hospital Clínic, CIBERehd, Barcelona, Spain;

2Department of

Immunology and Respiratory, Boehringer Ingelheim Pharmaceuticals Inc. Ridgefield Conne; 3Department of

Small Molecule Discovery Research, Boehringer Ingelheim Pharmaceuticals Inc. Ridgefiel; 4Department of

Gastroenterology, Hospital Universitari Mutua Terrassa, Terrassa, Barcelona, Spain; 5Centro de

Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid; 6Department of Gastroenterology, Hospital Universitari de Bellvitge- IDIBELL, Barcelona, Spain.

Increasing evidence suggests that Crohn's disease (CD) results from an aberrant immune

response to commensal microorganisms. Since deregulated pathways include Th17

responses, the inhibition of RORC represents a potential therapeutic strategy. Our aim

was to determine the ex vivo effect of RORC antagonism on circulating leukocytes and

intestinal tissue samples.

Peripheral blood mononuclear cells (PBMCs) from CD patients were cultured for 7 days

with commensal microbial proteins (FrvX and YidX) in the presence of a RORC antagonist.

We also examined the effects of commensal-specific CD4+ T cells treated with a RORC

inhibitor on healthy intestinal epithelial crypts.In addition, intestinal biopsies from active CD

patients were cultured with a RORC antagonist for 18h.

The RORC antagonist specifically inhibited transcription of Th17-related genes in bacterial

antigenstimulated PBMCs (n=12). Intestinal crypts cultured with supernatants from FrvX

(n=8) and YidX (n=7) specific CD4+ T cells previously treated with RORC inhibitor showed

decreased expression of CXCL1, CXCL8 and CCL20. Remarkably, biopsies from CD

patients treated with the RORC inhibitor significantly decreased transcription of IL-17A, IL-

17F and IL-26 in both colon (n=10) and ileum (n=8) of CD patients with active

inflammation. Other genes such as CSF2, CXCL1, CXCL8, IFNG, S100A8, IL-6 and

DEFA5 were also significantly regulated by RORc antagonist in ileal CD.

In conclusion, we demonstrate that blocking RORC specifically decreases the expression

of a subset of IL-17 dependent genes in the context of CD. This effect is observed in both

immune and epithelial cells, indicating that RORC antagonism could represent a

therapeutic approach for treating CD.

www.sci.cat 47

Posters



23 Diagnostic algorithm for the detection of anti-SOX1 antibodies

M. García Ormaechea1; M. Español-Rego

1; E. Martínez Hernández

2,3; L. Sabater

2; L. Querol

4; I. Illa

4; F.

Graus2,3

1Servei d’Immunologia, Centre Diagnòstic Biomèdic, Hospital Clínic de Barcelona;

2Neuroimmunology

Program, Institut d’Investigacions Biomèdiques August Pi i Sunyer, Barcelona; 3Servei de Neurologia,

Hospital Clínic de Barcelona; 4Unitat de Neuromuscular, Hospital de la Santa Creu i Sant Pau, Barcelona

Introduction: The Lambert-Eaton myasthenic syndrome (LEMS) is a neuromuscular

disorder with an associated cancer in up to 50% of patients, mostly lung cancer (LC). The

detection of anti-SOX1 antibodies is useful in the diagnosis of paraneoplastic LEMS,

although they had been detected sporadically in idiopathic neuropathies by

immunoblot/ELISA. Currently, there are commercial immunoblots for the detection of anti-

SOX1 antibodies but comparison with a specific technique is lacking and their sensitivity is

unknown.

Methods: We studied 203 serum samples using cell based assay (CBA), rat cerebellar

immunohistochemistry (IHQ) and commercial immunoblot (IB). We included 64 patients

with anti-SOX1 antibodies and 139 as disease controls (41 idiopathic sensitive

neuropathy, 49 chronic inflammatory demyelinating polyneuropathy, 18 paraneoplastic

neuropathy without LC and 31 anti-MAG positive monoclonal gammopathy).

Results: 79.7% anti-SOX1 positive patients were male. Mean age was 64 years. 98.4%

patients had cancer (61 lung, 1 prostate and 1 breast); 25% presented cerebellar

degeneration; 22% LEMS; 22% limbic encephalitis; 11% encephalomyelitis; 11%

neuropathy/neuronopathy; 3% Stiff-man syndrome; 6% LC without any neurological

syndrome and 2% cerebellar syndrome without cancer. 10.9% anti-SOX1 positive serum

samples by CBA were negative by IHQ but positive by IB (IHQ false negatives) and

another 10.9% were negative by IB (IB false negatives). Regarding the 139 controls, only

1 patient, with sensitive neuropathy and parotid cancer, was positive for SOX1 by IB but

seronegative by CBA and by IHQ (IB false positive). Anti- SOX1 abs were detected by all

three tests in 78% of patients.

Conclusions: Anti-SOX1 antibodies are LC markers. It is important to assess the risk of

LC in samples negative by IB (>40 years, smoking, classic paraneoplastic syndrome),

since 11% of patients have anti-SOX1 antibodies by CBA. False positive results are rare

and should be confirmed by CBA if clinical picture is discordant.

48 www.sci.cat

Posters



24 The Trex2 exonuclease and the DNase1L2 endonuclease

cooperate in the DNA degradation process during lingual

keratinocyte cornification

Joan Climent1,2

; Joan Manils1; Heinz Fischer

3; Eduard Casas

4; Celia García-Martínez

1; Jordi Bas

1,2;

Tanya Vavouri4; Supawadee Sukseree

3; Josep María de Anta

1; Erwin Tschachler

3; Leopold Eckhart

3;

Concepció Soler1

1Pathology and Experimental Therapeutics, Faculty of Medicine and Health Sciences, Universitat de

Barcelona; 2Immunology Dept, Hospital Universitari de Bellvitge;

3Research Division of Biology and

Pathobiology of the Skin, Medical University of Vienna, Austria; 4Institute Germans Trias i Pujol, Spain

Introduction: Cornification is a unique mode of programmed cell death in keratinocytes

undergoing terminal differentiation on the skin epidermis and other squamous epithelia. It

is associated with the physiological degradation of nuclear DNA. Delay or absence of this

DNA fragmentation and degradation events may result in an aberrant DNA-driven immune

response.

Objectives: Taking in account that Trex2 processes DNA from 3’-OH ends that can be

generated by DNase1L2 we hypothesized that both nucleases might cooperate in DNA

degradation during keratinocyte enucleation. Here, we investigated the effects in mice of

simultaneous loss of these two keratinocyte-specific nucleases in keratinocyte enucleation.

Methods: Histological and immunological studies in single (DNase1L2-/- or Trex2-/-) and

double (DNase1L2-/-Trex2-/-) deficient mice were perfomed to asses the extent of

fragmented-DNA retention and the DNA driven immunological response in keratinocyte-

built tissues, such as skin and tongue epithelia.

Results: Both DNases cooperated in the DNA degradation during keratinocyte

cornification in tongue epithelium, but were dispensable for homeostatic epidermal skin

cornification. Simultaneous loss of Trex2 and DNase1L2 led to a synergistic accumulation

of DNA fragments rather than a simple sum of single knockout phenotypes. The abnormal

accumulation of cytosolic DNA in lingual corneocytes was not associated to DNAdriven

immune responses, antinuclear autoantibody generation and inflammation.

Conclusions: 1) There is a tissuedependent interplay of Trex2 and DNase1L2 in

keratinocyte DNA degradation during lingual cornification. 2) Lingual epithelium displays

physiological tolerance to aberrantly retained endogenous DNA that may be explained in

part by low expression of key DNA sensing and signaling genes.

www.sci.cat 49

2018 Events

Lifelong Learning SCI Program 2018 Data i hora: 1 de febrer de 2018, a les 18:30h

“Virus del Papil·loma humà: Disseny de la vacuna, implantació i test per a

screening”

Dr. Jorge Lloberas (Departament de Biologia Cel·lular, Fisiologia i Immunologia,

Universitat de Barcelona), Dr. Xavier Bosch (Institut Català d’Oncologia), Dra. Cristina

Vanrell Barbat (Servei d’Obstetrícia i Ginecologia de l’Hospital de la Santa Creu i Sant

Pau, Barcelona). Auditori, Acadèmia de Ciències Mèdiques (Major de Can Caralleu 1-7,

Barcelona).

Data i hora: 1 de març de 2018, a les 18:30h

“Transcriptòmica del Lupus Eritematós Sistèmic”

Dra. Cristina Sole Marcé (Grup de Malalties Sistèmiques Autoimmunes de l'Institut de

Recerca Vall d'Hebron, Barcelona), Dra. Teresa Sardón (Anaxomics). Auditori, Acadèmia

de Ciències Mèdiques (Major de Can Caralleu 1-7, Barcelona).

Data i hora: 26 d'abril de 2018, a les 16:00h.

“Systems Biology”

Dr. Alex Sánchez Pla (Grup de Bioinformàtica de l'Institut de Recerca Vall d'Hebron,

Barcelona), Dr. Xavier de la Cruz Montserrat (Grup de Bioinformàtica Translacional a

l'Institut de Recerca Vall d'Hebron, Barcelona), Dr. Roger Colobran Oriol (Grup

d'Immunogenètica, Hospital Vall d'Hebron, Barcelona). Sala Pere i Joan Coromines de

l’Institut d'Estudis Catalans (C/ del Carme, 47, Barcelona).

Data i hora: 3 de maig de 2018, a les 18:30h.

“Gut Microbiome and Inflammatory Bowel Diseases”

Dra. Chaysavanh Manichanh (Fisiologia i Fisiopatologia del tracte digestiu, Institut de

Recerca Vall d’Hebron, Barcelona). Auditori, Acadèmia de Ciències Mèdiques (Major de

Can Caralleu 1-7, Barcelona).

Data i hora: 7 de juny de 2018, a les 18:30h.

“Situació actual de la Investigació en Tuberculosis i desenvolupament de noves

vacunes”

Dr. Carlos Martin Montañés (Universitat de Zaragoza), Dr. Pere Joan Cardona (Unitat

de Tuberculosi Experimental. Institut Germans Trias i Pujol, Badalona) Auditori, Acadèmia

de Ciències Mèdiques (Major de Can Caralleu 1-7, Barcelona).

50 www.sci.cat

New members

Registration form to SCI

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Participant information

Useful information

Congress Venue: Academia de Ciències Mèdiques, Auditori de l’Acadèmia c/ Major de Can Caralleu 1 08017 Barcelona. www.sci.cat Transportation:

By car: Ronda de Dalt, exit 9

By bus: o Line 66 (Pl. Catalunya – Sarrià) o Line 60 (Pl. Glòries – Zona Universitaria) o Line V3 (Zona Franca –Can Caralleu) o Line 130 (Pl. Artós – Can Caralleu))

By subway: Ferrocarrils de la Generalitat de Catalunya (FGC): Line 6: Reina Elisenda station Parking: Small open area between the Academy and the city ring (only for members of the Academy: Parking fees apply). Congress Office: Sr. Xavier Nieves Tel: 00 34 93.203.13.18 FAX: 00 34 93 212 35 69 [email protected]

http://www.sci.cat/

mailto:[email protected]

52 www.sci.cat

List of participants and authors

Abril-Gil M. 37, 40, 42, 44 Aguilera E. 14 Álvarez I. 19, 20 Ara del Rey J. 27 Aragon L. 25 Arajol C. 46 Aran A. 7, 23 Arribas J. 23 Arribas-Layton D. 19 Azagra-Boronat I. 34, 40 Badell I. 15 Barastegui R. 46 Barrachina S. 30, 32 Bas J. 13, 30, 31, 32, 48 Bassolas-Molina H. 46 Baucells de la Peña A. 6, 15 Bayes-Genis A. 17 Bel J. 29 Bernadó C. 23 Bernardi C. 39 Bernuz M. 23 Boada A. 26 Boera-Carnicero G. 15 Bordignon J. 31 Borràs F.E. 17, 24 Borràs N. 33 Bosio M. 22 Briansó F. 14 Calderón E. 8 Calderón-Gómez E. 46 Canals F. 20 Cano-Sarabia M. 14 Cantó E. 35, 36 Carrascal M. 20 Carrasco A. 46 Carreras-Planella L. 7, 24 Casas E. 48 Castell M. 34, 37, 40, 42, 44 Celada A. 18 Climent J. 48 Colobran R. 22, 25 Corral M. 21 Cortés J. 23 Costa C. 38 Cucurull M. 26 Dalmau J. 7 Daura X. 19 De Andrés B. 7 de Anta J.M. 48 De la Calle O. 15 de la Torre M.A. 28 del Mazo-Barbara A. 12 Domínguez-Alonso C. 25 Dotti I. 46 Eckhart L. 48 Egía-Mendikute L. 21, 28 Engel P. 7 Enrich E. 33 España S. 26 Español-Rego M. 47 Esteller M. 46

Esteve C. 26, 27 Esteve M. 46 Etxaniz O. 26 Fajas Ll. 21 Farriol R. 43 Fernández M.A. 45 Fernández-Prendes C. 26 Ferrer-Picón E. 46 Fillatreau S. 8 Fischer H. 48 Fonolleda M. 29 Franch A. 34, 37, 40, 42, 44 Franco-Jarava C. 22, 25 Franquesa M. 17, 24 Fuertes C. 30 Fulladosa X. 31 Gallego M. 46 Gálvez-Montón C. 17 García M. 7, 47 García M.J. 31 García-Jimeno S. 14 García-Martínez C. 48 García-Prat M. 22, 25 Garssen J. 34 Gimeno R. 7 Granada M.L. 26 Grases-Pintó B. 37, 40, 42, 44 Graterol F.A. 27 Graus F. 47 Grau-Vorster M. 6, 12 Guardiola J. 46 Guarner C. 35 Harcken C. 46 Hernández-Álvarez M. 18 Hervás S. 6 Holgado E. 23 Hughes R. 46 Illa I. 47 James E.A. 19 Juan M. 6 Juarez C. 6 Juárez C. 35, 41 Julián M.T. 45 Knipping K. 34 Knol J. 34 Kuguel S.G. 38 Labadia M. 46 Lloberas J. 18 López E. 16 López I. 21 López-Montañés M. 12 Manils J. 48 Mansilla M.J. 6, 10, 39 Marcilla M. 19 Mari A.V. 25 Marín-Morote L. 37 Marín-Sánchez A. 11 Mariscal-Rodríguez A. 36 Marqués M. 26 Martí M. 23 Martínez E. 47

www.sci.cat 53

Martínez-Cáceres E.M. 7, 10, 26, 27, 39, 45 Martínez-Gallo M. 22 Martínez-Martínez L. 15, 36 Martín-Nalda A. 22, 25 Mas V. 6, 13, 30, 31, 32 Masamunt M.C. 46 Maspoch D. 14 Massot-Cladera M. 34, 40 Mata E. 41 Mayol R. 13, 32 Mestre-Ferrer A. 20 Moga E. 41 Mongay L. 33 Monguió-Tortajada M. 7, 17, 24 Mora C. 21, 28 Morandeira F. 13, 30, 31, 32 Muñoz J.P. 18 Murillo M. 29 Nabozny G. 46 Navarro S. 6, 13, 30, 31, 32 Navarro-Barriuso J. 10, 39 Nieto J.C. 35 Oliver-Vila I. 12 Ortiz M.A. 36 Ossowski S. 22 Padró A. 32 Panés J. 46 Pariente-Cano A. 15 Peg V. 23 Perea L. 35, 36 Pereira-Lopes S. 18 Pérez A.C. 26 Pérez J. 23 Pérez-Cano F.J. 34, 37, 40, 42, 44 Perna-Barrull D. 14 Perurena-Prieto J. 25 Piron M. 33 Pizarro E. 45 Plaja A. 22 Planell N. 46 Poyatos E. 13, 30, 31, 32 Presas-Rodríguez S. 10, 39 Puig L. 41 Pujol-Autonell I. 14 Pujol-Borrell R. 2, 6, 8, 11 Querol L. 47 Querol S. 12, 33 Quirant B. 10, 26, 27, 39 Ramo-Tello C. 10, 39 Raymond E. 46 Regueiro J.R. 25 Richarz N. 26 Rivera N. 26

Rodríguez-Fernández S. 6, 14, 29 Rodríguez-Lagunas M.J. 34, 37, 40 Román E. 35 Rosell-Mases E. 21, 28 Roura S. 17 Rudilla F. 12, 33 Sabater L. 47 Saiz J. 31 Salas A. 46 Sánchez A. 14 Sánchez E. 35 Sánchez M.A. 41 Santamaria P. 16 Santos-Rosendo C. 7, 21, 28 Sanz M.T. 11, 41 Sarrias M.R. 7 Sauleda S. 33 Scholz E. 7, 19, 20 Serra P. 16 Smith D. 46 Soberino J. 23 Soler C. 48 Soler-Palacín P. 7, 22, 25 Soriano G. 35 Stahl B. 34 Stratmann T. 21 Sukseree S. 48 Teniente A. 10, 26, 27, 39, 45 Tims S. 34 Torres-Castro P. 42 Tschachler E. 48 Tur J. 7, 18 Turner M. 46 Unanue E. 18 Uribe-Herranz M. 38 Valledor A. 7 van’t Land B. 34 Vavouri T. 48 Vázquez F. 14, 29 Vega J. 7, 16 Verdaguer J. 6, 14, 21, 28 Vico T. 18 Vidal F. 33 Vidal S. 35, 36, 41 Vila-Pijoan G. 6, 11 Villalba A. 14, 29 Vived C. 21, 28 Vives J. 12 Vives-Pi M. 6, 14, 29 Wahle J. 46 Zamora C. 35, 36, 41 Zamora E. 23 Zorzano A. 18

54 www.sci.cat

Other useful information and notes

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www.sci.cat 55

XI CONGRÉS

Societat Catalana d’Immunologia (SCI)

Barcelona, 16 i 17 de novembre 2017

The contents of this congress will be accessible in a few months in our website www.congressci.com

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