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with persistent enteropathy and Pneumocystis jiroveci

pneumonia.
Methods. Whole exome sequencing (WES) was performed on
the patient. Flow cytometry was used to evaluate protein
expression and immunophenotyping.
Results. WES revealed a homozygous 1 base-pair deletion in ICOS causing a frameshift and premature truncation in exon 2, which was confirmed by Sanger
sequencing in the patient and his sister, who has
hypogammaglobulinemia, chronic diarrhea, low memory
B cells, and low CD4+CXCR5+CD45RO+ T follicular
helper (Tfh) cells. The mutation abolishes ICOS expression
in both patients. ICOS is needed for the development of
germinal center (GC)-associated CXCR5+ Tfh cells and GC
B cells. The patients enteropathy resolved after HSCT, a
novel finding indicating that his colitis was driven by hematopoietic cells. In contrast, his sister, who has remained
untransplanted, has persistent diarrhea.
Conclusion. Although this mutation caused classical features
of ICOS deficiency, including hypogammaglobulinemia, low
memory B cells, and low CXCR5+ Tfh cells, chronic
enteropathy and the probands opportunistic infection
suggest that ICOS deficiency is not solely a disease of
defective class-switching. The ICOS-ICOSL interaction
may be important for macrophage clearance of
P. jiroveci and a loss-of-function ICOSL polymorphism
is associated with Crohns ileitis. It is important to
consider antibiotic prophylaxis for opportunistic infections in ICOS-deficient patients and HSCT for those
with severe enteropathy.

Objective: To describe the immunological and molecular phenotype of a child with DGS-like syndrome born to a diabetic
mother.
Methods: Cytogenetic and molecular analysis includes fluorescent in situ hybridization (FISH), Array-CGH and TBX1
sequencing. Lymphocyte subpopulations were studied by
flow-cytometry and PBMC proliferation by standard method.
Results: a 6-year-old Caucasian male was admitted for thymic
aplasia, recurrent airway infections, hypoparathyroidism, renal agenesis, patent oval foramen, septum pellucidum cyst,
language delay and dysmorphisms. The immunological evaluation revealed normal serum Ig levels. The
immunophenotype revealed a reduction of CD3+ (373cell/
mm3), CD4+ (163cells/mm3), CD8+ (388cells/mm3), CD3+
CD4+CD45RA (47cells/mm 3 ), CD3+CD4+CD45RO
(116cells/mm 3 ) cells. B-lymphocytes were increased
(1352cells/mm3), while CD56+ were normal. The proliferative response to PHA and Pockweed was decreased, corresponding to the 45% and 39% of the control, respectively.
FISH analysis and TBX1 sequencing were negative.
Array CGH revealed a 371Kb-interstitial deletion at
3p12.3 involving the ZNF717, MiRNA-1243 and 4273
genes. Among the MiRNA-4273 predicted target-genes,
we found Bone Morphogenetic protein-3 (BMP-3), involved in several steps of embryogenesis, as in kidney and
lung organogenesis.
Conclusions: We report on a novel association between a DGlike phenotype and a 3p12.3 chromosomal deletion in an
infant born to a diabetic mother, although the causal relationship remains to be proved.

ESID-0200 Digeorge-Like Syndrome in a Child with a

3p12.3 Deletion Involving MiRNA-4273 Born to a Diabetic
Mother

ESID-0683 A Founder Splicing Mutation in CD3D in a

Tab-Tgd+B+NK+SCID Pedigree of Ecuadorian Descent

E. Cirillo , V. Gallo , G. Giardino , G. Galasso , R. Romano ,

R. D'Assante1, R. Genesio2, A. Baldini2, L. Nitsch2, C.
Pignata1

L. Martinez-Martinez1, R. Cedeo2, A.V. Marin3, M.V.

Rubiales1, A. Jimnez-Reinoso3, M.J. Recio3, P. Cabrera4, I.
Badell5, J.R. Regueiro3, O. de la Calle-Martn6
1

Immunology, Hospital de Sant Pau, Barcelona, Spain

Pediatrics, Hospital Oncolgico Dr. Julio Villacreces
Colmont, Portoviejo, Ecuador
3
Immunology, School of Medicine Complutense University,
Madrid, Spain
4
Cytometry, Cytolab, Cuenca, Ecuador
5
Pediatrics, Hospital de Sant Pau - Universitat Autonoma de
Barcelona, Barcelona, Spain
6
Immunology, Hospital de Sant Pau - Universitat Autonoma
de Barcelona, Barcelona, Spain

Introduction: Chromosome 22q11.2 deletion is the most commonly chromosomal alteration associated with DiGeorge syndrome (DGS). However, 22q11.2 deletion is not the only
underlying cause of DGS. Recently it has been reported that
miRNA may modulate the expression of critical T-box transcriptional regulators during midface development and the
Bmp-signalling.

Introduction: Severe Combined Immunodeficiency (SCID) is

a group of rare and fatal-inherited disorders characterized by

Translational Medical Sciences, Universit "Federico II",

Naples, Italy
2
Molecular Medicine and Medical Biotechnology, Universit
"Federico II", Naples, Italy

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early onset of infections and diminished T cell numbers and/or

strongly impaired T cell function.
Aims: To identify the genetic and molecular alteration in an
Ecuadorian patient with TB+NK+SCID.
Methods: The patient, a 6-month-old girl of
nonconsanguineous parents, presented with fever, eczema,
and respiratory distress. CMV pneumonia was diagnosed,
and active CMV infection also caused chorioretinitis. The
patient also presented regional lymphadenitis following
BCG vaccination (BCGitis). Lymphopenia, and severe
hypogammaglobulinemia were observed. Lymphocyte subpopulations were determined by flow cytometry. CD3 gamma
and delta genes were analyzed in DNA from patient and
parents's peripheral blood leukocytes.
Results: The patient showed a severe selective reduction in
peripheral blood T lymphocyte numbers (both CD4+ and
CD8+). In contrast, T cells as well as B and NK lymphocytes were detected in normal numbers (T-T+B+NK+
phenotype). TCR and CD3 expression levels were strongly
affected. Genomic DNA sequencing detected a homozygous
G-to-A mutation at position +5 in the 5' splice donor site of
intron 2 (IVS2+5G>A). The patients' parents were carriers of
the same CD3D mutation. This mutation has been previously
described in two non-related nonconsanguineous families of
Ecuadorian descent. The 3 families reported to date are unrelated, but had a similar geographic origin (Manab region),
indicating that they likely shared a frequent founder mutant
allele.
Conclusions: We report a TB+NK+SCID patient belonging
to a complex pedigree with a homozygous splicing mutation
in the CD3 delta gene (c.274+5G>A).
ESID-0414 The Role of STAT3 in the Differentiation
and Function of Unconventional T Cells
E.K. Deenick1, M.L. Ives1, R.P. Wilson1, G. Uzel2, S.G.
Tangye1
1

Immunology, Garvan Institute of Medical Research,

Darlinghurst, Australia
2
Laboratory of Clinical Infectious Diseases, NIAID NIH,
Bethesda, USA
Introduction: Unconventional or innate-like T cells include
T, natural killer T (NKT) and mucosal-associated invariant T
(MAIT) cells. MAIT and T cells are a major component of
the human immune system, comprising 1-15% of peripheral
blood lymphocytes and have been implicated in responses to
many pathogens. However, our knowledge of their precise
role in protective immunity and the molecular mechanisms
regulating their development, activation and function is

limited. Human primary immunodeficiencies caused by lossof-function mutations in single genes provide a unique opportunity to assess the requirement for particular molecules as
regulators of human unconventional T-cell activation and
differentiation.
Objective: To determine the role of STAT3 in regulating the
differentiation and function of unconventional T cells.
Methods: Autosomal dominant-hyper IgE syndrome
(AD-HIES) is caused by mutations in STAT3. We have
used lymphocytes from these patients to assess the roles
of STAT3 in NKT, MAIT and T cell differentiation
in vivo and in vitro.
Results: Patients with STAT3 mutations displayed normal numbers of T cells but reduced numbers of MAIT and NKT cells
ex vivo. Further, T cells and MAIT cells from these patients
also displayed altered responses to in vitro stimulation.
Conclusion: STAT3 plays an important role in regulating
unconventional T cells. Defects in these cells may contribute
to the clinical phenotype observed in AD-HIES.
ESID-0529 Opportunistic Infections in a Patient
with X-Men Disease
F. Dhalla1, S. Murray1, R. Sadler1, J. Miller1, E. Soilleux2, M.
Bhole3, B. Ferry1, H.M. Chapel1, S.Y. Patel1
1

Department of Immunology, Oxford University Hospitals

NHS Trust, Oxford, United Kingdom
2
Department of Histopathology, Oxford University Hospitals
NHS Trust, Oxford, United Kingdom
3
Department of Immunology, Russells Hall Hospital, Dudley,
United Kingdom
X-MEN disease (X-linked immunodeficiency with
Magnesium defect, Epstein-Barr virus infection and
Neoplasia) is a novel primary immune deficiency caused by
loss of function mutations in MAGT1, which encodes a selective magnesium transporter found to be important for T-cell
receptor signalling and for NKG2D expression on NK- and
CD8 T-cells. To date only 7 patients, ranging from 3-45 years
of age, have been described in the literature, identifying
chronic infection with Epstein-Barr virus (EBV), EBVdriven lymphoma, CD4 T-cell lymphopaenia, and
dysgammaglobulinaemia as clinical features. Here we
present the case of a 58 year-old man with MAGT1
deficiency and hope to add to the clinical phenotype of
this newly described primary immune deficiency by
naming Pneumocystis pneumonia and progressive multifocal leucoencephalopathy (PML) as possible complications, particularly in the context of treatment with chemotherapeutic agents and Rituximab.