Annual General Meeting of the Irish Thoracic Society - IJMS | Irish ...

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SESSION ONE
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ANNUAL MEETING OF THE IRISH THORACIC SOCIETY • 11 - 12 November 2005 • WESTWOOD HOUSE HOTEL, GALWAY ANNUAL MEETING OF THE IRISH THORACIC SOCIETY • 11 - 12 November 2005 • WESTWOOD HOUSE HOTEL, GALWAY
SESSION ONE
1.8
1.9
Macrophage Migration Inhibitory Factor (MIF) enhances
growth of P. aeruginosa in vitro: potential therapeutic strategy
in Cystic Fibrosis
Introduction
MIF is a pro-inflammatory factor involved in the
pathogenesis of inflammatory diseases. We have
demonstrated increased levels of MIF in plasma from
CF patients. MIF is a unique cytokine as a result of its
enzymatic activity, the role of which is undetermined.
Here, we examine the effect of MIF on P. aeruginosa
growth in vitro and the role of its enzymatic activity
on bacterial growth.
Method
P. aeruginosa (ATCC TM 27853) was cultured according
to standard methods in DMEM (Gibco BRL) in the
presence or absence of recombinant human MIF
(rhMIF; 1-100 ng/ml). The number of bacteria (CFU/
ml) was confirmed by serial dilution followed by
culture on LB plates. To investigate the role of the
enzymatic activity of MIF, P. aeruginosa was cultured
in conditioned media harvested from embryonic
fibroblasts from either MIF + / + , MIF - / - mice or mice
which were devoid of MIF enzymatic activity
(P1G mice).
Results
An increase in P. aeruginosa growth (CFU/ml) was
observed in the presence of rhMIF at (1ng/ml; 3.0x10 9
± 9.5 x 10 8 ) compared with media alone (1.69 x 10 9
± 2.59 x 10 8 ). Furthermore, a significant increase in P.
aeruginosa growth was observed following culture in
conditioned media from embryonic fibroblasts from
MIF+/+ mice (2.4X10 9 ±1.1 9 CFU/ml) compared with
mice that were devoid of MIF enzymatic activity (P1G
mice) (7.7X10 8 ±2.6x10 8 CFU/ml).
Conclusion
P. aeruginosa growth is enhanced in the presence of
rhMIF and this effect may be due to its enzymatic
activity. This novel observation suggests a potential
for anti-MIF strategies in the management of
pseudomonas infection in CF.
ADAM family of genes play a role in collagen deposition in
an in vitro model of pulmonary fibrosis
AM McLaughlin,
ME Armstrong,
JA Baugh, SD Donnelly
Dept of Medicine and
Therapeutics, The
Conway Institute of
Biomolecular and
Biomedical Research,
UCD and St Vincent’s
University Hospital,
ElmPark, Dublin
1.10
Safety and efficacy of prolonged treatment of fibrosing
alveolitis in scleroderma with cyclophosphamide
Background
Although cyclophosphamide has been widely
used for the treatment of fibrosing alveolitis in
Scleroderma, there has been debate over the optimal
treatment regime.
Methods
Two groups of patients were studied separately at
our centre. The first group (Group A, 14 patients)
were treated with 6 intravenous pulses of
cyclophosphamide and methylprednisolone at
monthly intervals for 6 months 1 . A second group of
patients (Group B, 26 patients) were treated with 15
pulses of cyclophosphamide and methylprednisolone
at intervals increasing from 3 to 12 weeks.
There was the option of increasing the dose of
cyclophosphamide if patients deteriorated during
treatment. Both groups were assessed with serial
pulmonary function tests and HRCT thorax.
Time
(months)
Results
Both groups of patients had similar characteristics in
terms of disease duration and initial lung function.
The effects of treatment on pulmonary function
tests are shown below.
Conclusion
Prolonged treatment with cyclophosphamide and
methylprednisolone appears safe and may be more
effective in reducing the progression of lung fibrosis
in scleroderma.
Reference
1. Griffiths B et al. Systemic sclerosis and
interstitial lung disease: a pilot study to assess
pulse intravenous methylprednisolone and
cyclophosphamide J Rhematol 2002; 29(11):2371.
GROUP A GROUP B GROUP A GROUP B
% Change
in DLCO
p-value
% Change
in DLCO
p-value
% Change
in FVC
p-value
Change in
FVC
p-value
6 -7.4 0.24 0.5 0.75 0 0.66 0 0.70
12 -8.6 0.39 -4.3 0.06 3.3 0.34 0.25 0.20
24 -10 0.04 -6.6 0.14 0.5 0.25 1.2 0.16
*P-values have been calculated by the Wilcoxon rank test.
*No patients withdrew from treatment because of toxicity from cyclophosphamide and no deaths were recorded.
1.11
MT Henry, 2 S Dass, 1
C Fernandes, 1
B Griffiths, 1 P Emery, 1
1. Dept of
Rheumatology, and
2. Dept of Respiratory
Medicine, Leeds
General Infirmary,
Leeds, UK
Introduction
The molecular mechanisms of Idiopathic Pulmonary
Fibrosis(IPF) remain elusive. Transforming Growth
Factor beta 1(TGF-ß1) is a key effector cytokine in the
development of lung fibrosis. We used microarray
and computational biology strategies to identify
gene clusters whose expression is significantly
altered in alveolar epithelial cells.
Method
A549 cells were exposed to 10ng /ml TGFß1 for serial
time points. Total RNA was used for hybridisation to
Affymetrix Human Genome U133A microarrays. Each
in vitro time-point was studied in triplicate and an
average RMA value computed. Expression data for
each time point was compared to control and a signal
log ratio of 0.6 or greater taken to identify significant
differential regulation. Using normalised RMA values
and unsupervised Average Linkage Hierarchical Cluster
Analysis, a list of 200 ECM proteins or modulators of
matrix turnover was curated via Onto-Compare
and Gene-Ontology(GO) databases for baited cluster
analysis of ECM associated genes.
Results
Interrogation of the dataset using ontological
classification focused cluster analysis revealed
coordinate differential expression of a large cohort
of extracellular matrix associated genes. Of this
grouping members of the ADAM (A disintegrin and
Metalloproteinase domain containing) family of
genes were found to be differentially expressed. We
probed pathologenomic activities (activation and
functional activity) of ADAM19 and ADAMTS9 using
siRNA and collagen assay.
Conclusion
Knockdown of ADAM genes resulted in diminished
production of collagen in A549 cells exposed to
TGFß1, suggesting a role for these molecules in ECM
accumulation in IPF.
DT Keating, 1, 2
A Patricelli, 1 D Sadlier, 1
P Doran, JJ Egan 2
1. Genome Research
Unit, and
2. Advanced Lung
Disease Programme,
Mater Misericordiae
Hospital, Dublin
Efficacy of pulmonary rehabilitation in interstitial lung disease
Background
Pulmonary rehabilitation (PRP) is effective in
improving exercise endurance and quality of life
(QoL) in COPD. However, data on the efficacy of PRP in
interstitial lung disease (ILD) is limited .
Methods
35 patients with ILD (mean DLCO 40.3±14.9%pred
, mean age 67.7±10.7) were admitted to PRP; 19
completed the 8 weeks programme and 10 were
followed to 1 year. Pulmonary function tests, exercise
endurance and QoL (Chronic Respiratory Disease
- CRDQ, St.George’s Respiratory - SGRQ, Hospital
Anxiety and Depression - HAD) and dyspnoea were
measured at baseline, 8 weeks and 1 year. The sample
studied was subgrouped - group 1 DLCO ≤35%pred.
(n=14), group 2 DLCO >35% pred. (n=17). 4 patients
were unable to perform the test.
Results
Overall exercise endurance (treadmill) improved
at 8 weeks (p