Læknablaðið : fylgirit - 03.01.2017, Page 34

Introduction: The neonatal immune system is immature. Antibody (Ab)
responses are slow and transient due to reduced survival of antibody-
-secreting cells (AbSCs) in the bone marrow (BM). The aim was to study
the effect of neonatal immunization w/wo adjuvant LT-K63 on BM cell
subsets and their production of the AbSC survival signals APRIL and IL-6.
Materials and methods: We assessed the frequency of neutrophils,
eosinophils (EOs), monocytes, macrophages and megakaryocytes
(MKCs), as well as APRIL- and IL-6-secreting cells in BM by FACS at
different time points after neonatal immunization with pneumococcal
conjugate (Pnc1-TT) w/wo the adjuvant LT-K63. Vaccine-specific AbSCs in
BM and serum Abs were measured by ELISPOT and ELISA.
Results: Frequency of MKCs and EOs, and APRIL- and IL-6-secreting
MKCs and EOs was significantly increased in mice immunized with Pnc1-
TT+LT-K63 compared to Pnc1-TT. Furthermore, the fraction of EOs that
were APRIL+ and IL-6+ and MKCs that were APRIL+ was larger in mice
immunized with Pnc1-TT+LT-K63. This correlates with higher vaccine-
-specific Ab levels and number of AbSCs that persist in the BM of mice
that receive LT-K63 together with the vaccine.
Conclucion: The results suggest that the adjuvant LT-K63 not only incre-
ases the frequency of EOs and MKCs in BM of neonatal mice but also act-
ivates a higher percentage of EOs to secrete APRIL and IL-6 and a higher
percentage of MKCs to secrete APRIL. This might contribute to increased
survival of AbSCs in BM at early age when LT-K63 is administered to-
gether with Pnc1-TT.
E 89 Endotheliomics: The metabolic response of the endothelium to
LPS and hypoxia
Sarah McGarrity1, Ósk Anuforo2, Haraldur Halldórsson3, Óttar Rolfsson3
1School of Engineering and Natural Sciences, Center for Systems Biology, 2Center for Systems
Biology, 3Faculty of Medicine, School of Health Sciences
sarahm@hi.is
Introduction: Endothelial cells line blood vessels and their dysfunction
plays a key role in many diseases, especially inflammatory conditions
including sepsis. Context specific genome scale metabolic models
(GEMs) have been built to explore metabolic changes in different endot-
helial cell types when grown with lipopolysaccharide (LPS) and under
hypoxia. These models combine existing transcriptomic data with novel
metabolomic data.
Methods: Using the COBRAtoolbox and the Fastcore algorithm the
RECON1 model was constrained to produce three cell type specific
GEMs related to human umbilical vein, pulmonary artery and
microvascular endothelial cells (HUVEC, HPAEC and HMVEC) using
publically available transcriptomic data. The HUVEC model was further
constrained to reflect HUVECs grown with LPS and the HPAEC model
to reflect cells grown under hypoxia. Metabolomic data from HUVECs
with and without LPS was collected using mass spectrometry and app-
lied to further constrain the model. Comparisons between models were
made; analysing the differences in lethal reactions between models and
MOMA to analyse changes in growth rates.
Results: Preliminary analysis of cell type specific GEMs show small
differences between endothelial cell types in glycolysis and amino
acid metabolism between different endothelial cell types. Differences
in nucleotide metabolism and pyruvate metabolism appear between
HUVECs with and without LPS and HPAECs grown under normoxic
and hypoxic conditions show differences in pyruvate metabolism and
amino acid metabolism.
Conclusions: Combining metabolomics analysis with models based on
transcriptomics data highlights context specific differences endothelial
cell metabolism that may have functional implications.
E 90 Clarifying the role of signal transduction of keratinocytes in the
pathogenesis of psoriasis
Hildur Sigurgrímsdóttir1,2, Fannar P. Theódórs1,2, Jóna Freysdóttir1,2,3, Björn R.
Lúðvíksson1,2
1Faculty of Medicine, Biomedical Center, University of Iceland, 2Dept of Immunology,
Landspitali – The National University Hospital of Iceland, 3Centre for Rheumatology Research,
Landspitali – The National University Hospital of Iceland
hildursigur@hotmail.com
Introduction: Psoriasis is a chronic inflammatory skin disease, character-
ized by infiltration of T cells and epidermal thickening. Several cytokines
and anti-microbial peptides are upregulated in psoriasis patients, inclu-
ding IFNγ, IL-17, IL-22 and TNFα and together with immune cells in-
filtrating the skin they drive the pathogenesis of psoriasis. Phsophoflow
is a new way of measuring intra-cellular signaling pathways by flow
cytometry. It provides advantages over the older method, western
blotting, as it can measure both the percentage of activated cells within a
homo- or heterogeneous population and the mean expression level.
The main aim of this research is to characterize the intracellular response
of keratinocytes to different stimuli using phosphoflow.
Material and methods: HaCaT cells (immortalized keratinocytes)
stimulated with selected cytokines were stained for phosphorylated
intracellular proteins and analyzed using flow cytometry.
Results: Using phosphoflow, phosphorylation of cytokine-dependent
signaling molecules in HaCaT cells could be detected. The response to
IFNγ, the main Th1 cytokine, was characterized by phosphorylation of
STAT1 while the main cytokines in a Th17 response, IL-17 and IL-22,
induced an increase in ERK1/2 phosphorylation. IL-4, which directs Th2
differentiaion, induced phosphorylation of STAT6.
Conclusion: This study shows that phosphoflow can be used to inve-
stigate keratinocyte signaling pathways which may shed further light on
how pro-inflammatory cytokines and/or anti-microbial peptides influence
keratinocytes and subsequently the pathogenesis of psoriasis.
E 91 Innleiðing og notagildi Klínískra leiðbeininga um hjúkrun
heilablóðfallsjúklinga í endurhæfingu
Ingibjörg Bjartmarz1, Helga Jónsdóttir2, Þóra B. Hafsteinsdóttir3
1Endurhæfingardeild á Grensás, 2Hjúkrunarfræðideild, Heilbrigðisvísindasvið, 3Department of
Rehabilitation, Nursing Science and Sport, Brain Centre Rudolf Magnus, University Medical
Center Utrecht
ingibjart@simnet.is
Inngangur: Tilgangur rannsóknarinnar var að meta innleiðingu og hag-
nýta notkun “Klínískra leiðbeininga um hjúkrun heilablóðfallssjúklinga í
endurhæfingu” sem fela í sér ráðleggingar um hreyfi- og sjálfsbjargargetu,
þunglyndi og fræðslu. Einnig var metin reynsla hjúkrunarfræðinga og
sjúkraliða af innleiðingu og notkun leiðbeininganna.
Efniviður og aðferðir: Klínísku leiðbeiningarnar á formi skriflegra,
myndrænna, netupplýsinga og veggspjalda voru innleiddar á taugalækn-
ingadeild og endurhæfingardeild Grensás á Landspítala. Eftirfarandi ferli
var notað við innleiðinguna: a) valdir voru lykilhjúkrunarfræðingar og
sjúkraliðar til að styðja við innleiðinguna; b) fræðslufundir voru haldnir
með öllu starfsfólki; c) hjúkrunarfræðingar og sjúkraliðar fengu kennslu
og starfsþjálfun. Til þess að meta áhrif innleiðingarinnar á skráningu og
34 LÆKNAblaðið/Fylgirit 91 2017/103
X V I I I V Í S I N D A R Á Ð S T E F N A H Í
F Y L G I R I T 9 1