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Protective role of M-CSF dependent macrophages in experimental colitis in mice
Abstracts
99
97
EFFICACY OF PANTOPRAZOLE IN HEALING P.!ZFLUX OESOPHAGITIS BOTH IN PRESENCE AND ABSENCE OF HELICOBACTER PYLORI. Bazzoli F, Zagari R.M., Pozzato P, Fossi S, Nicolbd G, De Luca L, BcrretU D, Ricciardiello L, Marh& C, Maltoni S, Fuccio L, Roda E On behalf of Pantoprazole
Study Group. Dept. of Internal Medicine and Gastmenterology, University of Bologna, Italy
lmtroduction. The relation between Helicobac~erpylm (Hp.) infection and GORD is still unclear. The thetapctdic success of the r&secretory dmgs in healing reflw ocsophaf$is has been related to their capability of increasing intragas~ic PH. It has been shown that intragashic pH is higher during PPIs therapy in Rp.+ve than H.p.-ve patients, the presence of H p. infection could increase the efticacy of PPIs in the treatment of r&.x oesophagitis. Aim. To evahtate the efticacy of Pantoprazole 40 mg o.d. in the treatment of refhlx oesophagitis both in presettce or in absence 0fH.p. infection. Methods. 301 patients (61.I%malea; meanage 46.1+ 15.4 yrs, range 19-83 yrs) with rethx oesophagitis ( Grade I-III Savary-Miller) were enrolled in a multlcentre, randomlsed, double-blind, controlled study for acute therapy with Pantoptazole 40 mg o.d. for 8 weeks and maintettance therapy with pantopramle. Upper GI cndoscopy was &wfortned before and after the end of the therapy. At entry, H.p. status (histology, RUT and “C-urea breath test) was also established. Patients with H.p. infestion were randomixd to receive µbial treatment with clarithromycin 250 mg b.d + dnidazole 500 nu b.d. for 1 week. or not treatment. GORD svmotoms were evaluated before and after iweeks and 8 weeks therapy. Results. 286 &t if 301 patients completed the study and were evaluated At 2 motuhs~&erall, 96.2% of patients were healed (94% of those made I oesofaeitis at entrv. 98.5% of those made II and 90% of those made III, At entw-32.2 % of oat&s
was H.b.+ve. Oesooha&s
healing rate was not i&emxdbv
H.p. S&S or by an&H.p. treatm&t (96.7% H.b. -ie, 95.9%l?p.+ve
and 94.3% in thosk
who received antibiotic txatment). 74.3% and 84.8% of patients were GORD symptoms free after 4 weeks and 8 weeks of treatnerd. Also GORD svmotoms imorovement was independent by H.p. status. Conclusions. Pantoprazole 40 &g’o.d for s’ weeks 1shighly etfective in healing t’etlux oesophagitis. The H. &ori status or antimicrobial treatment do not influence the efficacy of PPIS therapy. This research was funded by Pharmacia & Upjohn, Mllq
Italy
PROTECTIVE ROLE OF MCSF DEPENDENT MACROPHAGES IN EXPERIMENTAL COLITIS IN MICE. F Galeazzi’~‘, C.M. Hogabwn’, B A Valiance’, GC Stumiolo”2. S.M Collins’ ‘IDRP and G I Division, M&faster University & HHSC-Hamilton CANADA; *Department of Surgical and Gastmenterological Science, IJmverslty of Padova Italy, ‘Dew ofpatholonv. Universitv ofMichinan. Ann Arbor. Michigan, Ma&phages pl& a critim~ role in-intestinal infl&mati& Indeed, blockade of n&ophage-dewed pro-inflammatory cytokines has been shown to be beneficial in IBD and in animal models However. this cell type contribute to host dcfence and immune responses Thus, urn of thw study was to investigate the role of macrophages in experimental colitis m mice, using mice deficient in macrophages as a result of a mutation in the macmphage-colony stimulating factor (MCSF) encoding gene METHODS M-CSF-deficient mice (ophp) as well as their M-CSF expressing littermates (op/+) received intrarectally 3 mg of dinitrobenzensulfonic acid @NBS) in 50% ethanol Mice were euthamzed at day I and day 3 post colitis induction and were evaluated macroscopic damage of the colon, myeloperoxidase activity (MPO), presence of macroohaaes .- bv. immunohistochemisbv for F4/80 and levels of the chemokine MI&la, involved in the recruitment of inflammatory cells, by ELISA In parallel experiments, mice received daily intr&riton&l iniections of human recombinant M-CSF Ihr MCSF) RESULTS In op/~ rmce, DNBS-induced whtls was accompanied by a massive transmural macrophagic infiltrate, whde in M-CSF deficwt mice only few F4/80 positive cells were evident. The severity of colitis was greater in M-CSF deficient mice compared to op/i mice, both at day 1 and day 3 post induction (macroscopic score day I op/+. 4439, op/op 7 2?0 8, day 3 op 3 3+0 5, op/op 7 3+0 5), and MPO activity was signiticantly higher in ophp mice at day 1 post c&is (op/+ I *to 3, op’op 3 7il 0) Coliu was accompanied increased MlP-la levels, which were IO times lugher in ophp over op/+ mice both at day I and at day 3 post mlitls Treatment with hrM-CSF restored the macrophagic infiltrate, reversed the increase in M?‘O activity observed in op/op mice, and amelioratedthe macroscopx damage at day 3 post colitis. CONCLUSIONS M-CSF-dependent macrophages play a protectwe role 1” hapten-inducedcahtis in mice This is likely due to a local overpraductmn of m-la, leading to an exaggerated neutrofilic infiltration, as shown by MF’O levels, wlxch is likely responsible far the greater damage These data suggest that macrophages are wt only a source of proinflammatory mediators, but they might play a role in maintuning mucosal defences and limiting the severity of mflammstion.
98 ROLE OF ANTIMICROBIAL EFFICACY OF TRIPLE ERADICATION.
SUSCEPTIBILITY TESTING ON THERAPY IN Helicobacter pylon’
S L Cellini’, A Ferri, MG Malatesta, G Cappello, AP Ciccaglione, E Di Campli’, L Grossi, L Marzio. G. D’Annunzio University of Chieti; * Department of Biomedical Sciences; School of Gastroenterology c/o Pierangeli Clinic, Pescara; Italy. Introduction: Amoxicillin (AMOXI), clarithromycin (CLA) and tmidazole gw) or metronidazole (METRO) are the most frequently used antibacteriaJ drugs in the therapy afH pylori eradication. Treatment failure seems to be due to resistance of H pyhito macrolides and S-nitroimidazoles Aim: ‘IIK aim ofthis study has been to test whether a preliminary “in vitro” susceptibility test of H. pylori to TIN and CLA and a consequent specific therapy could improve the eradication rate inpatients affected by H. pyh infection. Materials and methods: A total of 109 consecutive patients with idiopathic dyspepsia without any prior eradication treatment and a positive 13CUreaBreath Test were included. At endoscopy, biopsy f?omantrum was obtained for H. pyloti culh~re and antimicrobial susceptibility testing. 53 patients (Group A) recewed therapy after antibiogram and 56 patients (Group B) were treatedatoncewithOME20mgb.i.d.,TIN500mgb.i.d.andCLA500mgb.i.d. for 10 days. Susceptibility testing for TIN and CLA was performed by screening agar method and by the agar dilution method. Strains were ConsideredTIN and CLA resistant ifh4IC was L 5 &ml and L 1 p&l, respectively. Patients in group A were treated withOMEZOmgb.~.d,TIN500mgb.i.d. andCLA500mgb.i.d. ifH. pylori was sensible to both antibiotics, OME 20 mg b.i.d., TIN 500 mg b.i.d. and AMOXI 1 g b.i.d. if H. pyhi was resistant to CLA; OME 20 mg b.i.d., AMOXI 1 g b.i.d. and CLA 500 mg b.i.d. if R pyhi was resistant to TIN; OME 20 trig b.i.d. and AMOXI 1 g b.i.d. ifH. pylori was resistant to both antibiotics. Treatment
success was evaluated by the “C Urea Breath Test one month after the end of therapy. Results: Eight patients dropped out Overall primary resistance to clarithromycin, tinidazole and both antibiotics was 12.9%, 32.7% and 4%, respecttvely. One month after the end of treatment, in group B eradication was achieved in 80.8% and 75% of patients by per protocol (PP) and intention-to-treat @l-f) analysis. respectively. PP and ITr eradication rates for group A were 97.9% and 90.6% (P < 0.05 vs group B). Conclusions: These data show that in H. pyhi mfectmn, antlbiotlc therapy based on the results of culture and susceptibiity testing gwes, in comparison to standard therapy, a significant improvement in eradication rate
SERUM CHONDREX ASSOCIATED WITH Ferronato A, ‘Pozzuoli ‘Bernardi
D, D’Incl
(YLK-40) IN PATIENTS WITH ARTHRITIS INFLAMMATORY BOWEL DISEASE (IBD) A, “Pumi L, “Podswiadek M, *Mcneghetti F,
R, ‘Plebani
M, Stumiolo
GC.
Dep. Surgical and Gastroenterological Sciences, “Dep. of Internal Medicine, *Laboratory Unit, University of Padua, Padua, Italy Background. Crohn’s disease (CD) and Ulcerative colitis (UC) are chronic inflammatory bowel diseases which sometimes show extraintestinal manifestations to the joints. Biochemical markers of inflammation are often aspecific and it is sometimes difficult to discriminate between activity of the disease per se and arthroparhic activity. YKL-40 is a 40 kD glycoprotein mainly secreted by chondrocytes and synoviocytes. Its serum concentrations have been found to be elevated in patients with inflammatory atthropathies and correlated with laboratory indices ofjoints inflammation. Aims. To investigate the clinical value of serum YKL-40 in characterising, among patients with IBD, those having arthritis (IBD-A). Methods. YKL-40 levels were assayed in the serum of 43 patients with IBD-A, 23 Crohn’s disease (CD) and 20 ulcerative colitis (UC), 16 with IBD without arthritis (IBDnonA) and 20 normal subjects, as controls (C). C-reactive protein (CRP)was assayed in the serum of the same subjects and patients, as an index of acute phase response. YKL-40 levels were determined by ELISA (Metra Biosystem, USA), CRP by nephelometry. Results. Serum levels of YKL-40 were significantly higher in IBD-A patients (140.3+109.7 rig/ml) with respect lo IBDnonA patients (54.0+18.9 “g/ml, p=O.OOl) and C (55.9k16.7 @ml, p=O.OOl). No difference was found in YKL-40 serum levels between IBDnonA and C or, among the subgroups, between CD (150.7f114.6 rig/ml) and CU (128.3+105.4 n&l). A significant correlation was found between YKL-40 and CRP (t=O.512, p=O.OOOl). Conclusions. The increased serum levels of YKL-40 and its correlarion with CRP found in IBD-A suggest that YKL-40 may be an useful marker of arthritis associated with IBD.
A89
99
97
EFFICACY OF PANTOPRAZOLE IN HEALING P.!ZFLUX OESOPHAGITIS BOTH IN PRESENCE AND ABSENCE OF HELICOBACTER PYLORI. Bazzoli F, Zagari R.M., Pozzato P, Fossi S, Nicolbd G, De Luca L, BcrretU D, Ricciardiello L, Marh& C, Maltoni S, Fuccio L, Roda E On behalf of Pantoprazole
Study Group. Dept. of Internal Medicine and Gastmenterology, University of Bologna, Italy
lmtroduction. The relation between Helicobac~erpylm (Hp.) infection and GORD is still unclear. The thetapctdic success of the r&secretory dmgs in healing reflw ocsophaf$is has been related to their capability of increasing intragas~ic PH. It has been shown that intragashic pH is higher during PPIs therapy in Rp.+ve than H.p.-ve patients, the presence of H p. infection could increase the efticacy of PPIs in the treatment of r&.x oesophagitis. Aim. To evahtate the efticacy of Pantoprazole 40 mg o.d. in the treatment of refhlx oesophagitis both in presettce or in absence 0fH.p. infection. Methods. 301 patients (61.I%malea; meanage 46.1+ 15.4 yrs, range 19-83 yrs) with rethx oesophagitis ( Grade I-III Savary-Miller) were enrolled in a multlcentre, randomlsed, double-blind, controlled study for acute therapy with Pantoptazole 40 mg o.d. for 8 weeks and maintettance therapy with pantopramle. Upper GI cndoscopy was &wfortned before and after the end of the therapy. At entry, H.p. status (histology, RUT and “C-urea breath test) was also established. Patients with H.p. infestion were randomixd to receive µbial treatment with clarithromycin 250 mg b.d + dnidazole 500 nu b.d. for 1 week. or not treatment. GORD svmotoms were evaluated before and after iweeks and 8 weeks therapy. Results. 286 &t if 301 patients completed the study and were evaluated At 2 motuhs~&erall, 96.2% of patients were healed (94% of those made I oesofaeitis at entrv. 98.5% of those made II and 90% of those made III, At entw-32.2 % of oat&s
was H.b.+ve. Oesooha&s
healing rate was not i&emxdbv
H.p. S&S or by an&H.p. treatm&t (96.7% H.b. -ie, 95.9%l?p.+ve
and 94.3% in thosk
who received antibiotic txatment). 74.3% and 84.8% of patients were GORD symptoms free after 4 weeks and 8 weeks of treatnerd. Also GORD svmotoms imorovement was independent by H.p. status. Conclusions. Pantoprazole 40 &g’o.d for s’ weeks 1shighly etfective in healing t’etlux oesophagitis. The H. &ori status or antimicrobial treatment do not influence the efficacy of PPIS therapy. This research was funded by Pharmacia & Upjohn, Mllq
Italy
PROTECTIVE ROLE OF MCSF DEPENDENT MACROPHAGES IN EXPERIMENTAL COLITIS IN MICE. F Galeazzi’~‘, C.M. Hogabwn’, B A Valiance’, GC Stumiolo”2. S.M Collins’ ‘IDRP and G I Division, M&faster University & HHSC-Hamilton CANADA; *Department of Surgical and Gastmenterological Science, IJmverslty of Padova Italy, ‘Dew ofpatholonv. Universitv ofMichinan. Ann Arbor. Michigan, Ma&phages pl& a critim~ role in-intestinal infl&mati& Indeed, blockade of n&ophage-dewed pro-inflammatory cytokines has been shown to be beneficial in IBD and in animal models However. this cell type contribute to host dcfence and immune responses Thus, urn of thw study was to investigate the role of macrophages in experimental colitis m mice, using mice deficient in macrophages as a result of a mutation in the macmphage-colony stimulating factor (MCSF) encoding gene METHODS M-CSF-deficient mice (ophp) as well as their M-CSF expressing littermates (op/+) received intrarectally 3 mg of dinitrobenzensulfonic acid @NBS) in 50% ethanol Mice were euthamzed at day I and day 3 post colitis induction and were evaluated macroscopic damage of the colon, myeloperoxidase activity (MPO), presence of macroohaaes .- bv. immunohistochemisbv for F4/80 and levels of the chemokine MI&la, involved in the recruitment of inflammatory cells, by ELISA In parallel experiments, mice received daily intr&riton&l iniections of human recombinant M-CSF Ihr MCSF) RESULTS In op/~ rmce, DNBS-induced whtls was accompanied by a massive transmural macrophagic infiltrate, whde in M-CSF deficwt mice only few F4/80 positive cells were evident. The severity of colitis was greater in M-CSF deficient mice compared to op/i mice, both at day 1 and day 3 post induction (macroscopic score day I op/+. 4439, op/op 7 2?0 8, day 3 op 3 3+0 5, op/op 7 3+0 5), and MPO activity was signiticantly higher in ophp mice at day 1 post c&is (op/+ I *to 3, op’op 3 7il 0) Coliu was accompanied increased MlP-la levels, which were IO times lugher in ophp over op/+ mice both at day I and at day 3 post mlitls Treatment with hrM-CSF restored the macrophagic infiltrate, reversed the increase in M?‘O activity observed in op/op mice, and amelioratedthe macroscopx damage at day 3 post colitis. CONCLUSIONS M-CSF-dependent macrophages play a protectwe role 1” hapten-inducedcahtis in mice This is likely due to a local overpraductmn of m-la, leading to an exaggerated neutrofilic infiltration, as shown by MF’O levels, wlxch is likely responsible far the greater damage These data suggest that macrophages are wt only a source of proinflammatory mediators, but they might play a role in maintuning mucosal defences and limiting the severity of mflammstion.
98 ROLE OF ANTIMICROBIAL EFFICACY OF TRIPLE ERADICATION.
SUSCEPTIBILITY TESTING ON THERAPY IN Helicobacter pylon’
S L Cellini’, A Ferri, MG Malatesta, G Cappello, AP Ciccaglione, E Di Campli’, L Grossi, L Marzio. G. D’Annunzio University of Chieti; * Department of Biomedical Sciences; School of Gastroenterology c/o Pierangeli Clinic, Pescara; Italy. Introduction: Amoxicillin (AMOXI), clarithromycin (CLA) and tmidazole gw) or metronidazole (METRO) are the most frequently used antibacteriaJ drugs in the therapy afH pylori eradication. Treatment failure seems to be due to resistance of H pyhito macrolides and S-nitroimidazoles Aim: ‘IIK aim ofthis study has been to test whether a preliminary “in vitro” susceptibility test of H. pylori to TIN and CLA and a consequent specific therapy could improve the eradication rate inpatients affected by H. pyh infection. Materials and methods: A total of 109 consecutive patients with idiopathic dyspepsia without any prior eradication treatment and a positive 13CUreaBreath Test were included. At endoscopy, biopsy f?omantrum was obtained for H. pyloti culh~re and antimicrobial susceptibility testing. 53 patients (Group A) recewed therapy after antibiogram and 56 patients (Group B) were treatedatoncewithOME20mgb.i.d.,TIN500mgb.i.d.andCLA500mgb.i.d. for 10 days. Susceptibility testing for TIN and CLA was performed by screening agar method and by the agar dilution method. Strains were ConsideredTIN and CLA resistant ifh4IC was L 5 &ml and L 1 p&l, respectively. Patients in group A were treated withOMEZOmgb.~.d,TIN500mgb.i.d. andCLA500mgb.i.d. ifH. pylori was sensible to both antibiotics, OME 20 mg b.i.d., TIN 500 mg b.i.d. and AMOXI 1 g b.i.d. if H. pyhi was resistant to CLA; OME 20 mg b.i.d., AMOXI 1 g b.i.d. and CLA 500 mg b.i.d. if R pyhi was resistant to TIN; OME 20 trig b.i.d. and AMOXI 1 g b.i.d. ifH. pylori was resistant to both antibiotics. Treatment
success was evaluated by the “C Urea Breath Test one month after the end of therapy. Results: Eight patients dropped out Overall primary resistance to clarithromycin, tinidazole and both antibiotics was 12.9%, 32.7% and 4%, respecttvely. One month after the end of treatment, in group B eradication was achieved in 80.8% and 75% of patients by per protocol (PP) and intention-to-treat @l-f) analysis. respectively. PP and ITr eradication rates for group A were 97.9% and 90.6% (P < 0.05 vs group B). Conclusions: These data show that in H. pyhi mfectmn, antlbiotlc therapy based on the results of culture and susceptibiity testing gwes, in comparison to standard therapy, a significant improvement in eradication rate
SERUM CHONDREX ASSOCIATED WITH Ferronato A, ‘Pozzuoli ‘Bernardi
D, D’Incl
(YLK-40) IN PATIENTS WITH ARTHRITIS INFLAMMATORY BOWEL DISEASE (IBD) A, “Pumi L, “Podswiadek M, *Mcneghetti F,
R, ‘Plebani
M, Stumiolo
GC.
Dep. Surgical and Gastroenterological Sciences, “Dep. of Internal Medicine, *Laboratory Unit, University of Padua, Padua, Italy Background. Crohn’s disease (CD) and Ulcerative colitis (UC) are chronic inflammatory bowel diseases which sometimes show extraintestinal manifestations to the joints. Biochemical markers of inflammation are often aspecific and it is sometimes difficult to discriminate between activity of the disease per se and arthroparhic activity. YKL-40 is a 40 kD glycoprotein mainly secreted by chondrocytes and synoviocytes. Its serum concentrations have been found to be elevated in patients with inflammatory atthropathies and correlated with laboratory indices ofjoints inflammation. Aims. To investigate the clinical value of serum YKL-40 in characterising, among patients with IBD, those having arthritis (IBD-A). Methods. YKL-40 levels were assayed in the serum of 43 patients with IBD-A, 23 Crohn’s disease (CD) and 20 ulcerative colitis (UC), 16 with IBD without arthritis (IBDnonA) and 20 normal subjects, as controls (C). C-reactive protein (CRP)was assayed in the serum of the same subjects and patients, as an index of acute phase response. YKL-40 levels were determined by ELISA (Metra Biosystem, USA), CRP by nephelometry. Results. Serum levels of YKL-40 were significantly higher in IBD-A patients (140.3+109.7 rig/ml) with respect lo IBDnonA patients (54.0+18.9 “g/ml, p=O.OOl) and C (55.9k16.7 @ml, p=O.OOl). No difference was found in YKL-40 serum levels between IBDnonA and C or, among the subgroups, between CD (150.7f114.6 rig/ml) and CU (128.3+105.4 n&l). A significant correlation was found between YKL-40 and CRP (t=O.512, p=O.OOOl). Conclusions. The increased serum levels of YKL-40 and its correlarion with CRP found in IBD-A suggest that YKL-40 may be an useful marker of arthritis associated with IBD.
A89