Download P036 Involvement of gut macrophages in the fibrogenic process in

Basic science
S25
Figure 2. IL-6 secretion by RAW 264.7 cells were stimulated for
24 h with LPS.
Table 2
Group
Weight
loss
Stool
consistency
Visible blood
in feces
Total
DAI score
Control*
AMA
Calcein**
DLT***
RO 41 0960
4
3
4
3
4
4
4
12
3
6
3
6
2
2
DAI, Disease Activity Index.
*5/6, **4/6 and ***2/6 animals die during the UC’s protocol at day 8.
Conclusions: According to the results, 3 out of 4 compounds
were able active in vitro, and 2 of them (Ro 41 0960 and DLT)
had also been revealed as a preventive compounds in vivo.
Reference(s)
[1] Atreya, R.; Neurath, M. F. (2005), Involvement of IL-6 in
the pathogenesis of inflammatory bowel disease and colon
cancer, Clin Rev Allerg Immu
[2] Gálvez-Llompart, M.; Recio, M. C.; García-Domenech,
R. (2011), Topological virtual screening: a way to find
new compounds active in ulcerative colitis by inhibiting
NF-kappaB., Mol. Divers
[3] Recio, M. C.; Gálvez-Llompart, M.; Zanni, R; GarcíaDomenech, R. (2011), New Inhibitors of Il-6 Production
In Caco-2 Cells Through Molecular Topology Methodology,
XXXIII Congreso de la Sociedad Española de Farmacología,
Málaga (Spain).
P036
Involvement of gut macrophages in the fibrogenic process
in Crohn’s disease
P. Biancheri1 *, A. Di Sabatino1 , I. Joe-Njoku2 , S.A. Snoek2 ,
C. Salvatore1 , P. Giuffrida1 , A. Ghanbari2 , T.T. MacDonald2 ,
G.R. Corazza1 . 1 Fondazione IRCCS Policlinico S. Matteo,
University of Pavia, First Department of Medicine, Italy,
2
Blizard Institute, Barts And The London School Of Medicine
And Dentistry, Centre for Immunology and Infectious Disease,
London, United Kingdom
Background: Intestinal fibrosis and stricture formation are a
frequent complication of Crohn’s disease (CD). Cytokine driven
excessive extracellular matrix deposition and an imbalance
between matrix metalloproteinases (MMPs) and their inhibitors
are thought to play an important role in the transmural process
of stricture development. We have therefore investigated the
immunological changes occurring in the deeper layers of the
bowel wall in CD patients and control subjects.
Methods: Surgical specimens were collected from strictured
and non-strictured ileum of 10 CD patients and from normal
ileum of 10 control subjects. The mucosa was removed
and discarded. Immunohistochemistry on muscle layers was
performed using anti-CD3, -CD4, -CD8, -HLA-DR, and -CD68
antibodies. Explants of strictured, non-strictured CD and
control gut were cultured ex vivo for 24 h. MMP-3 expression
and collagen, interleukin (IL)-1beta, IL-6 and tumor necrosis
factor (TNF)-alpha levels were measured in organ culture
supernatants.
Results: HLA-DR+ cells and macrophages (CD68+ cells) were
significantly increased in intestinal CD strictures as compared
to non-strictured CD and control gut. No significant differences
were found in total T (CD3+), CD4+, and CD8+ cell frequency
among the three groups of patients, and in general T cells were
infrequent. Collagen and IL-1beta were significantly increased
in organ culture supernatants from strictured CD as compared
to those from non-strictured CD and control gut. IL-6, TNFalpha and MMP-3 expression did not differ between strictured,
non-strictured CD and control gut supernatants.
Conclusions: The increased frequency of CD68+ cells in CD
strictures and the high levels of IL-1beta in strictured CD
supernatants suggest that macrophages may play an important
role in driving the processes of fibrogenesis in CD.
P037
Is ulcerative colitis an atypical Th2-mediated disease
characterised by excess production of interleukin-13?
P. Biancheri1 *, A. Di Sabatino1 , F. Ammoscato2 , I. Joe-Njoku2 ,
N. Ahmad2 , M. Guerci1 , P. Giuffrida1 , G.R. Corazza1 ,
T.T. MacDonald2 . 1 Fondazione IRCCS Policlinico S. Matteo,
University of Pavia, First Department of Medicine, Italy,
2
Blizard Institute, Barts And The London School Of Medicine
And Dentistry, Centre for Immunology and Infectious Disease,
London, United Kingdom
Background: Interleukin (IL)-13 is produced mostly by T helper
cell type (Th)2 cells and, together with IL-5, it has been
supposed to be a major cytokine implicated in sustaining the
pro-inflammatory immune response in ulcerative colitis. We
have therefore assessed IL-13 and IL-5 production by inflamed
mucosa and isolated lamina propria mononuclear cells (LPMCs)
from ulcerative colitis patients, and evaluated the effect of
IL-13 or its neutralization on the intestinal immune response in
inflammatory bowel disease.
Methods: Biopsies and LPMCs from the inflamed colon of 11
ulcerative colitis patients and 9 Crohn’s disease patients, and
from normal colon of 15 control subjects were cultured ex vivo
with or without anti-CD3/CD28-antibodies. IL-13, IL-5, IL-17
and interferon (IFN)-gamma production was measured by ELISA
in the cell and organ culture supernatants. Anti-CD3/CD28stimulated LPMCs were also cultured with recombinant human
IL-13 or with an anti-IL-13 neutralizing antibody, and the
production of IFN-gamma, IL-17 and tumor necrosis factor
(TNF)-alpha was determined by ELISA.
Results: Ex vivo IL-13 production did not differ between
ulcerative colitis, Crohn’s disease and control biopsies,
although IFN-gamma and IL-17 were significantly higher in
ulcerative colitis and Crohn’s disease than in controls. AntiCD3/CD28 stimulation induced a small increase in IL-13
production without significant differences between the three
groups, whereas IFN-gamma and IL-17 production by LPMCs was
significantly higher after anti-CD3/CD28 stimulation. IL-5 was
undetectable in all the organ culture or LPMC supernatants.
The addition of recombinant IL-13 or the neutralization of
endogenous IL-13 did not induce any significant change in IFNgamma, IL-17A and TNF-alpha production by anti-CD3/CD28stimulated ulcerative colitis and Crohn’s disease LPMCs.
Conclusions: Our findings do not support a prominent role for
IL-13 and the Th2 response in driving mucosal inflammation in
ulcerative colitis.