Francesca Paola Giugliano
@amc.nl
Tytgat Institute for liver and intestinal research
Amsterdam UMC
RESEARCH INTERESTS
Pediatrics
Gastroenterology
Intestinal maturation
IBD
Scopus Publications
Scopus Publications
Sarah Ouahoud, Francesca Giugliano, and Vanesa Muncan
Bio-Protocol, LLC
The measurement of transepithelial electrical resistance across confluent cell monolayer systems is the most commonly used technique to study intestinal barrier development and integrity. Electric cell substrate impedance sensing (ECIS) is a real-time, label-free, impedance-based method used to study various cell behaviors such as cell growth, viability, migration, and barrier function in vitro. So far, the ECIS technology has exclusively been performed on cell lines. Organoids, however, are cultured from tissue-specific stem cells, which better recapitulate cell functions and the heterogeneity of the parent tissue than cell lines and are therefore more physiologically relevant for research and modeling of human diseases. In this protocol paper, we demonstrate that ECIS technology can be successfully applied on 2D monolayers generated from patient-derived intestinal organoids. Key features • We present a protocol that allows the assessment of various cell functions, such as proliferation and barrier formation, with ECIS on organoid-derived monolayers. • The protocol facilitates intestinal barrier research on patient tissue-derived organoids, providing a valuable tool for disease modeling.
Ikrame Aknouch, Inés García-Rodríguez, Francesca Paola Giugliano, Carlemi Calitz, Gerrit Koen, Hetty van Eijk, Nina Johannessson, Sjoerd Rebers, Lieke Brouwer, Vanesa Muncan,et al.
Frontiers Media SA
Enterovirus A71 (EV-A71) can elicit a wide variety of human diseases such as hand, foot, and mouth disease and severe or fatal neurological complications. It is not clearly understood what determines the virulence and fitness of EV-A71. It has been observed that amino acid changes in the receptor binding protein, VP1, resulting in viral binding to heparan sulfate proteoglycans (HSPGs) may be important for the ability of EV-A71 to infect neuronal tissue. In this study, we identified that the presence of glutamine, as opposed to glutamic acid, at VP1-145 is key for viral infection in a 2D human fetal intestinal model, consistent with previous findings in an airway organoid model. Moreover, pre-treatment of EV-A71 particles with low molecular weight heparin to block HSPG-binding significantly reduced the infectivity of two clinical EV-A71 isolates and viral mutants carrying glutamine at VP1-145. Our data indicates that mutations in VP1 leading to HSPG-binding enhances viral replication in the human gut. These mutations resulting in increased production of viral particles at the primary replication site could lead to a higher risk of subsequent neuroinfection.ImportanceWith the near eradication of polio worldwide, polio-like illness (as is increasingly caused by EV-A71 infections) is of emerging concern. EV-A71 is indeed the most neurotropic enterovirus that poses a major threat globally to public health and specifically in infants and young children. Our findings will contribute to the understanding of the virulence and the pathogenicity of this virus. Further, our data also supports the identification of potential therapeutic targets against severe EV-A71 infection especially among infants and young children. Furthermore, our work highlights the key role of HSPG-binding mutations in the disease outcome of EV-A71. Additionally, EV-A71 is not able to infect the gut (the primary replication site in humans) in traditionally used animal models. Thus, our research highlights the need for human-based models to study human viral infections.
Ikrame Aknouch, Adithya Sridhar, Eline Freeze, Francesca Paola Giugliano, Britt J van Keulen, Michelle Romijn, Carlemi Calitz, Inés García-Rodríguez, Lance Mulder, Manon E Wildenberg,et al.
Life Science Alliance, LLC
Human milk is important for antimicrobial defense in infants and has well demonstrated antiviral activity. We evaluated the protective ability of human milk against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in a human fetal intestinal cell culture model. We found that, in this model, human milk blocks SARS-CoV-2 replication, irrespective of the presence of SARS-CoV-2 spike-specific antibodies. Complete inhibition of both enveloped Middle East respiratory syndrome coronavirus and human respiratory syncytial virus infections was also observed, whereas no inhibition of non-enveloped enterovirus A71 infection was seen. Transcriptome analysis after 24 h of the intestinal monolayers treated with human milk showed large transcriptomic changes from human milk treatment, and subsequent analysis suggested that ATP1A1 down-regulation by milk might be of importance. Inhibition of ATP1A1 blocked SARS-CoV-2 infection in our intestinal model, whereas no effect on EV-A71 infection was seen. Our data indicate that human milk has potent antiviral activity against particular (enveloped) viruses by potentially blocking the ATP1A1-mediated endocytic process.
Flora Fedele, Massimo Martinelli, Caterina Strisciuglio, Pasquale Dolce, Francesca Paola Giugliano, Elena Scarpato, Annamaria Staiano, and Erasmo Miele
Ovid Technologies (Wolters Kluwer Health)
Objectives: The aims of our study were to evaluate health-related quality of life (HRQoL) in children affected by inflammatory bowel disease (IBD) during the first wave of Coronavirus disease 2019 (COVID-19) pandemic and after 12 months. Methods: This was a single-center, prospective, observational study conducted between April 2020 and April 2021. Children from 10 to 18 years with a confirmed diagnosis of IBD were enrolled during the first COVID-19–related national quarantine. The following information was collected at the baseline and after 12 months: IBD subtype, location and phenotype, disease activity, current and previous therapies. Patients were asked to complete the PROMIS Anxiety and IMPACT III questionnaires. Results: One hundred and eighteen patients were enrolled, of whom 54 (46%) were affected by Crohn disease (CD) and 64 (54%) with ulcerative colitis (UC; median age: 15.5 years, range 10.3–18; M/F: 68/50). Median HRQoL was significantly decreased after 12 months compared with the beginning of COVID-19–related quarantine (T1: 76.7 vs T2: 72.8; P < 0.001). At 12 months, a higher number of children were reported to be in active disease when compared with the enrollment [T2: 22/108 (20.4%) vs T1: 12/118 (10%); P = 0.02]. Multivariate analysis showed a significant influence on HRQoL of quarantine period (P < 0.001), female sex (P = 0.016), biologic therapy (P = 0.011), and active disease (P < 0.001). Conclusions: A deterioration of HRQoL after 12 months from COVID-19–related quarantine was observed. Additionally, the higher number of children with active disease at 12 months compared with enrollment may suggest detrimental consequences of the reduced disease control, contributing to decreased HRQoL.
Marileen M. C. Prins, Francesca P. Giugliano, Manon van Roest, Stan F. J. van de Graaf, Pim J. Koelink, and Manon E. Wildenberg
The Company of Biologists
ABSTRACT The ATG16L1 T300A single-nucleotide polymorphism (SNP) is associated with Crohn's disease and causes an autophagy impairment. We have previously shown that this SNP is involved in the migration and hyperactivation of Rac1 in dendritic cells. Mucosal healing, currently the main target for inflammatory bowel disease treatment, depends on restoration of the epithelial barrier and requires appropriate migration of epithelial cells towards and over mucosal lesions. Therefore, we here further investigated the impact of autophagy on epithelial migration. ATG16L1 knockdown was established in the HT29 human colonic epithelial cell line using lentiviral transduction. Migratory capacity was evaluated using scratch assays and RhoAGTP was measured using G-LISA. Immunofluorescent ARHGAP18 and sequestome 1 (SQSTM1; also known as p62) staining was performed on HT29 cells and primary colonic tissue of Crohn's disease patients. We observed that ATG16L1 knockdown cells exhibited decreased autophagy and decreased migration capacity. Furthermore, activity of RhoA was decreased. These characteristics were phenocopied using ATG5 knockdown and pharmacological inhibition of autophagy. The migration defect was dependent on accumulation of SQSTM1 and was alleviated upon SQSTM1 knockdown. Strikingly, thiopurines also mitigated the effects of impaired autophagy. RhoA dysregulation appeared mediated through accumulation of the upstream regulator ARHGAP18, which was observed in cell lines, human foetal organoids and primary colonic tissue. Our results indicate that the ATG16L1 T300A Crohn's disease-associated SNP causes a decrease in migration capacity in epithelial cells, mediated by an increase in SQSTM1 and ARHGAP18 protein and subsequent reduced RhoA activation.
Francesca Paola Giugliano, Erasmo Miele, and Annamaria Staiano
Springer International Publishing
Massimo Martinelli, Francesca Paola Giugliano, Caterina Strisciuglio, Vaidotas Urbonas, Daniela Elena Serban, Aleksandra Banaszkiewicz, Amit Assa, Iva Hojsak, Tereza Lerchova, Víctor Manuel Navas-López,et al.
Oxford University Press (OUP)
Abstract Background Vaccine-preventable diseases and opportunistic infections in pediatric inflammatory bowel disease (IBD) are increasingly recognized issues. The aims of this study were to evaluate vaccinations, immunization status, and consequent therapeutic management in children with IBD and to analyze the differences among patients diagnosed before (Group 1) and after June 2012 (Group 2). Methods This was a multicenter, retrospective cohort investigation. Between July 2016 and July 2017, 430 children with IBD were enrolled in 13 centers. Diagnosis, therapeutic history, vaccinations, and immunization status screening at diagnosis and at immunosuppressant (IM)/biologic initiation and reasons for incomplete immunization were retrieved. Results Vaccination rates at diagnosis were unsatisfactory for measles, mumps, and rubella (89.3%), Haemophilus influenzae (81.9%), meningococcus C (23.5%), chickenpox (18.4%), pneumococcus (18.6%), papillomavirus (5.9%), and rotavirus (1.9%). Complete immunization was recorded in 38/430 (8.8%) children, but specific vaccines were recommended in 79/430 patients (18.6%), without differences between the 2 groups. At IM start, 22% of children were tested for Epstein-Barr virus (EBV) status, with 96.2% of EBV-naïve patients starting azathioprine, without differences between Groups 1 and 2. Screening for latent tuberculosis (TB) before start of biologics was performed in 175/190 (92.1%), with up to 9 different screening strategies and numerous inconsistencies. Conclusions We demonstrated a poor immunization status at diagnosis in children with IBD, which was not followed by proper vaccination catch-up. EBV status before IM initiation and latent TB before biologics were not adequately assessed. Thus, the overall impact of the current guidelines seems unsatisfactory.
C. Strisciuglio, E. Scarpato, S. Cenni, M.R. Serra, F.P. Giugliano, C.G. Mainolfi, P. Dolce, M. Martinelli, A. Staiano, and E. Miele
Elsevier BV
BACKGROUND
The relationship between exclusive enteral nutrition (EEN) and bone status is poorly defined in pediatric Crohn disease (CD).
AIMS
The aim of this study was to investigate the impact of EEN on body composition, nutritional status, and bone mineral density (BMD) in an incident CD cohort.
METHODS
18 newly diagnosed CD children starting EEN for 8 weeks were prospectively enrolled and evaluated at baseline and after 8 (T8), 26 (T26) and 52 weeks (T52) from diagnosis. The Fat Free Mass (FFM) and the Resting Energy Expenditure (REE) were measured through Bioelectrical Impedance (BIA) and the BMD was assessed by dual-energy X-ray (DXA). We compared DXA data of IBD patients to the data obtained in 15 healthy controls.
RESULTS
CD children had a significant lower BMD compared to healthy control both at baseline (p<0.0001), and after EEN therapy at T52 (p=0.0004); although at this latest time point CD children had a significant increase of BMD compared to baseline (p=0.0015). The BIA analysis showed a significant increase at T26 and T52 of FFM and REE. T52. FFM measured by BIA and BMD measured by DXA were significantly correlated.
CONCLUSION
EEN improves nutritional status and bone mineral composition.
Bartolomeus J. Meijer, Francesca P. Giugliano, Bart Baan, Jonathan H.M. van der Meer, Sander Meisner, Manon van Roest, Pim J. Koelink, Ruben J. de Boer, Nic Jones, Wolfgang Breitwieser,et al.
Elsevier BV
Background & Aims Activation factor-1 transcription factor family members activating transcription factors 2 and 7 (ATF2 and ATF7) have highly redundant functions owing to highly homologous DNA binding sites. Their role in intestinal epithelial homeostasis and repair is unknown. Here, we assessed the role of these proteins in these conditions in an intestine-specific mouse model. Methods We performed in vivo and ex vivo experiments using Villin-CreERT2Atf2fl/flAtf7ko/ko mice. We investigated the effects of intestinal epithelium-specific deletion of the Atf2 DNA binding region in Atf7-/- mice on cellular proliferation, differentiation, apoptosis, and epithelial barrier function under homeostatic conditions. Subsequently, we exposed mice to 2% dextran sulfate sodium (DSS) for 7 days and 12 Gy whole-body irradiation and assessed the response to epithelial damage. Results Activating phosphorylation of ATF2 and ATF7 was detected mainly in the crypts of the small intestine and the lower crypt region of the colonic epithelium. Under homeostatic conditions, no major phenotypic changes were detectable in the intestine of ATF mutant mice. However, on DSS exposure or whole-body irradiation, the intestinal epithelium showed a clearly impaired regenerative response. Mutant mice developed severe ulceration and inflammation associated with increased epithelial apoptosis on DSS exposure and were less able to regenerate colonic crypts on irradiation. In vitro, organoids derived from double-mutant epithelium had a growth disadvantage compared with wild-type organoids, impaired wound healing capacity in scratch assay, and increased sensitivity to tumor necrosis factor-α–induced damage. Conclusions ATF2 and ATF7 are dispensable for epithelial homeostasis, but are required to maintain epithelial regenerative capacity and protect against cell death during intestinal epithelial damage and repair.
Marialuisa Andreozzi, Francesca P. Giugliano, Teresa Strisciuglio, Elisabetta Pirozzi, Alfonso Papparella, Angela M. Caprio, Erasmo Miele, Caterina Strisciuglio, and Pasquale Perrone Filardi
Ovid Technologies (Wolters Kluwer Health)
OBJECTIVES
Chronic inflammation plays a central role in the etiology of endothelial damage. Endothelial dysfunction is the inability of the artery to dilate in response to an endothelial stimulus. We assessed the endothelial dysfunction (ED) by measuring the reactive hyperaemia index (RHI) and the flow-mediated dilation (FMD) in a cohort of pediatric patients affected by inflammatory bowel disease (IBD) and comparing these parameters to a group of healthy controls (HC).
METHODS
Forty-one patients were consecutive enrolled. ED was evaluated by both the plethysmographic RHI method and the measurement of the FMD of brachial artery after occlusion of the blood flow. Differences between patients and controls were assessed by the Mann-Whitney test. In each IBD patient, the main inflammation markers were detected and correlated to RHI and FMD by a linear regression test.
RESULTS
We enrolled 26 (59%) IBD patients and 18 (41%) HC. When comparing FMD value at diagnosis it was significantly lower in IBD patients than in HC (p = 0.04). This result was confirmed at follow-up, when this difference became even more significant (p = 0.004). A significant indirect correlation was found between FMD and fecal calprotectin (r:0.17;p = 0.04). No differences were found when comparing RHI.
CONCLUSIONS
Our results suggest that inflammation could lead to endothelial dysfunction assessed by ultrasound flow-mediated dilation (FMD). These data were not confirmed by RHI, however this could be due to the lack of a standardized pediatric cut-off. More studies are necessary to confirm our data.
Caterina Strisciuglio, Sabrina Cenni, Francesca Paola Giugliano, Erasmo Miele, Grazia Cirillo, Massimo Martinelli, Alessandra Vitale, Carlo Tolone, Annamaria Staiano, Emanuele Miraglia Del Giudice,et al.
Ovid Technologies (Wolters Kluwer Health)
Objectives: Existing studies usually do not measure the free vitamin D in pediatric patients with inflammatory bowel disease (IBD) and not consider the effect of inflammation on vitamin D levels. The aim of our study was to evaluate the concentrations of vitamin D–binding protein (VDBP), total and free 25-hydroxyvitamin-D (25(OH)D), and to correlate these values with the disease activity markers. Methods: Newly diagnosed children with IBD and a group of healthy controls (HCs) were enrolled. VDBP and total and free 25(OH)D levels were measured by enzyme-linked immunosorbent assay and compared using the Student t test. In each patient with IBD, the activity scores of disease and the main inflammation markers were correlated to total and free 25(OH)D levels. C-reactive protein was also measured in the control group, and it was related to VDBP by a linear regression test for all the groups. Results: Fifty-one consecutive children were enrolled: IBD = 33, HC = 18. Levels of total 25(OH)D were higher in HC than in patients with IBD (P = 0.01). The free/total 25(OH)D ratio was, however, higher in patients with IBD compared to HC (P < 0.001). Finally, levels of VDBP were lower in patients with IBD than in HC (P = 0.001). A significant direct correlation was found between the free/total 25(OH)D ratio and the activity index of disease (r2: 0.17; P = 0.01). Moreover, in patients with IBD and controls we found a significant indirect correlation between VDBP and C-reactive protein (r2: 0.12; P = 0.01). Conclusions: Inflammation inversely correlates to VDBP concentrations and patients with IBD, despite their deficiency in total 25(OH)D, have normal or even higher levels of free 25(OH)D.
Massimo Martinelli, Francesca P. Giugliano, Marina Russo, Eleonora Giannetti, Marialuisa Andreozzi, Dario Bruzzese, Laura Perrone, Annamaria Staiano, Emanuele Miraglia Del Giudice, Erasmo Miele,et al.
Ovid Technologies (Wolters Kluwer Health)
Objectives: The aims of this retrospective study were to describe ulcerative colitis (UC) phenotype at diagnosis and follow-up and to identify possible predictors of severe disease course. Methods: This was a retrospective, single-center study. We reviewed the charts of patients with UC diagnosed between 2 and 18 years at our referral center from January 2007 to January 2016. Laboratory and clinical features at diagnosis, such as disease extent, atypical phenotypes, extraintestinal manifestations, and therapies, and pattern changes during the follow-up, including relapse rate, disease extension, and the cumulative risk for colectomy were collected. Results: One hundred eleven patients were enrolled. Atypical phenotypes were identified at diagnosis in 55 out of 111 patients (49.5%). Extraintestinal manifestations were detected in 16 out of 111 (14.4%) at the diagnosis. During the follow-up 60 out of 111 (54%) patients needed to start azathioprine, 9 out of 111 (8.1%) patients started biologic therapy and 10 out of 111 (patients underwent surgery, resulting in a cumulative risk of 8% at 5 years and 16% at 10 years. Steroid refractoriness (hazard ratio: 13.9) and starting of biologic therapy (hazard ratio: 25.3) represented the best predictors for surgery. The cumulative probability of first relapse was 47% at 6 months and 63% at 1 year. Disease extension was reported in 21 out of 70 patients (30%). Conclusion: Pediatric UC is associated with a severe phenotype and a high percentage of atypical features. Surgery rate seems to be decreased from early reports.
Francesca Paola Giugliano, Caterina Strisciuglio, Massimo Martinelli, Marialuisa Andreozzi, Sabrina Cenni, Severo Campione, Maria D’Armiento, Annamaria Staiano, and Erasmo Miele
Elsevier BV
BACKGROUND
The new concept of disease remission for pediatric inflammatory bowel diseases (IBD) implies the achievement of mucosal healing.
AIMS
We aimed to evaluate endoscopic and histologic healing in children with Ulcerative Colitis (UC) and Crohn's disease (CD) in clinical remission after 52 weeks of Azathioprine.
METHODS
From December 2012 to July 2015 we prospectively enrolled IBD children starting Azathioprine. Enrolled patients in clinical remission underwent colonoscopy after 52 weeks. Macroscopic assessment was described with Mayo score and the simplified endoscopic score for UC and CD, respectively. For microscopic assessment, an average histology score was used. Data on inflammatory markers and fecal calprotectin were also collected.
RESULTS
Fourty-seven patients were included in the analysis. Endoscopic healing was detected in 20/26 (76.9%) UC children and 10/21 (47.6%) CD patients. Median Mayo score and simplified endoscopic score were significantly decreased at week 52 (p<0.001; p=0.005). Median average histology score was not significantly different at week 52 in both diseases. Fecal calprotectin was directly correlated with simplified endoscopic score (T0: r=0.4, p=0.05; T52: r=0.5, p=0.01), but not with Mayo score. No correlation was found between endoscopic and histologic scores.
CONCLUSIONS
IBD children under Azathioprine reach endoscopic healing, but not histological remission.
M. Martinelli, D. Ummarino, F. P. Giugliano, E. Sciorio, C. Tortora, D. Bruzzese, D. De Giovanni, I. Rutigliano, S. Valenti, C. Romano,et al.
Wiley
Infant colic (IC) is a prevalent physiological event of infants, which can disrupt the child's home environment. We aimed to investigate the effectiveness of a mixture of Matricariae chamomilla L., Melissa officinalis L. and tyndallized Lactobacillus acidophilus (HA122) compared with Lactobacillus reuteri DSM 17938 and with simethicone for the treatment of IC.
Caterina Strisciuglio, Francesca Giugliano, Massimo Martinelli, Sabrina Cenni, Luigi Greco, Annamaria Staiano, and Erasmo Miele
Ovid Technologies (Wolters Kluwer Health)
Objectives: The primary role of environment on inflammatory bowel disease (IBD) onset has been recently stressed. We aimed to investigate the effect of environmental factors in an IBD pediatric cohort. Methods: A total of 467 subjects (264 IBD and 203 controls) were enrolled. All patients underwent a questionnaire including 5 different groups of environmental risk factors: family history of IBD and autoimmune diseases, perinatal period, home amenities and domestic hygiene, childhood diseases and vaccinations, and diet. Results: In a multivariate model, mother's degree (odds ratio [OR]: 5.5; 2.5–11.6), duration of breast feeding >3rd month (OR: 4.3; 1.6–10.5), father's employment (OR: 3.7; 1.2–8.7), gluten introduction <6th month (OR: 2.8; 1.5–5), number of siblings <2 (OR: 2.8; 1.5–5.3), and family history of autoimmune diseases (OR: 2.7; 1.4–5.3) were significant risk factors for Crohn disease. Low adherence to Mediterranean diet (OR: 2.3; 1.2–4.5), gluten introduction <6th month (OR: 2.8; 1.6–4.9), and number of siblings <2 (OR: 2; 1.1–3.6) were significant risk factors for ulcerative colitis. Owning pets (OR: 0.3; 0.1–0.7) and bed sharing (OR: 0.2; 0.1–0.6) were protective factors for Crohn disease, whereas owning pets (OR: 0.4; 0.2–0.8) and family parasitosis (OR: 0.07; 0.01–0.4) were protective factors for ulcerative colitis. Conclusions: Our study confirms that environmental factors are closely linked to IBD onset and may partly explain IBD rise in developed countries.
Marina Russo, Francesca Paola Giugliano, Paolo Quitadamo, Valeria Mancusi, Erasmo Miele, and Annamaria Staiano
Springer Science and Business Media LLC
BackgroundAbout 30% of constipated children continue to struggle with constipation beyond puberty. Growing interest has recently raised on the use of probiotics as complementary therapy for FC, in order to prevent the possible PEG-related intestinal dysbiosis. Our study aimed at evaluating the effect on childhood FC of a probiotic mixture (PM), including Bifidobacteria breve M-16 V®, infantis M-63®, and longum BB536®.MethodsFifty-five consecutive children suffering from FC were randomly assigned into two groups: group A received a daily oral combination of PEG plus PM and group B received oral PEG only. Physical and clinical data were collected from each patient at week-1, week-2, week-4, and week-8.ResultsAfter 1 month, children who experienced improvement in the PEG and in the PEG + PM group were 88 and 81.8%, respectively (p = 0.24). After 1 month from the end of the study treatment, a positive trend towards a higher rate of clinical remission was observed within children treated with PM compared to those who took only PEG (percentage of children off therapy: 64 vs 52, respectively; p = 0.28).ConclusionsPEG and PEG + PM are equally effective and safe in the treatment of children with chronic constipation. Nevertheless, further studies are needed to show if adding Bifidobacteria strains to conventional therapy may lead to a better long-term outcome.
Caterina Strisciuglio, Erasmo Miele, Francesca P. Giugliano, Serena Vitale, Marialuisa Andreozzi, Alessandra Vitale, Maria R. Catania, Annamaria Staiano, Riccardo Troncone, and Carmen Gianfrani
Oxford University Press (OUP)
Abstract:Bifidobacteria have been reported to reduce inflammation and contribute to intestinal homeostasis. However, the interaction between these bacteria and the gut immune system remains largely unknown. Because of the central role played by dendritic cells (DCs) in immune responses, we examined in vitro the effects of a Bifidobacteria mixture (probiotic) on DC functionality from children with inflammatory bowel disease. DCs obtained from peripheral blood monocytes of patients with Crohn's disease (CD), ulcerative colitis, and noninflammatory bowel disease controls (HC) were incubated with fluorochrome-conjugated particles of Escherichia coli or DQ-Ovalbumin (DQ-OVA) after a pretreatment with the probiotic, to evaluate DC phenotype, antigen sampling and processing. Moreover, cell supernatants were collected to measure tumor necrosis factor alpha, interferon gamma, interleukin 17, and interleukin 10 production by enzyme-linked immunosorbent assay. DCs from CD children showed a higher bacteria particles uptake and DQ-OVA processing after incubation with the probiotic; in contrast, DC from both ulcerative colitis and HC showed no significant changes. Moreover, a marked tumor necrosis factor alpha release was observed in DC from CD after exposure to E. coli particles, whereas the probiotic did not affect the production of this proinflammatory cytokine. In conclusion, the Bifidobacteria significantly improved the antigen uptake and processing by DCs from patients with CD, which are known to present an impaired autophagic functionality, whereas, in DCs from ulcerative colitis and HC, no prominent effect of probiotic mixture was observed. This improvement of antigen sampling and processing could partially solve the impairment of intestinal innate immunity and reduce uncontrolled microorganism growth in the intestine of children with inflammatory bowel disease.
Caterina Strisciuglio, Renata Auricchio, Massimo Martinelli, Annamaria Staiano, Francesca Paola Giugliano, Marialuisa Andreozzi, Marina De Rosa, Eleonora Giannetti, Carmela Gianfrani, Paola Izzo,et al.
Elsevier BV
BACKGROUND AND AIMS
Little evidence demonstrating the correlation between several single nucleotide polymorphisms and a specific phenotype of Crohn's disease has been reported in children. We investigated the relationship between autophagy genes variants and clinical features in our children with Crohn's disease.
METHODS
Genotyping for ATG16L1, NOD2/CARD15, and IRGM1 was performed in 80 consecutive patients with Crohn's disease (median age: 11 years; range: 0.7-17.9 years). Crohn's disease location and behaviour were classified using the Paris classification. Additional data were collected from clinical records on patients' demographics, age at symptom onset and diagnosis, extraintestinal manifestations, therapy, clinical relapses, and need of surgical intervention.
RESULTS
Patients homozygous for the risk allele ATG16L1 (T300A) showed a trend towards switching to a stricturing phenotype during the course of disease compared to children either homozygous for the wild-type allele or heterozygous for the ATG16L1 single nucleotide polymorphism (p=0.01). Homozygosity for the ATG16L1 risk allele was associated with a major recurrence of clinical relapses and earlier introduction of immunosuppressants (p=0.006 and p=0.04, respectively). Heterozygosity for the NOD2 rs2066847 allele was associated with major ileal involvement (p=0.01).
CONCLUSION
In patients carrying the T300A variant, Crohn's disease follows a more aggressive clinical course.
Caterina Strisciuglio, Erasmo Miele, Manon E. Wildenberg, Francesca P. Giugliano, Marialuisa Andreozzi, Alessandra Vitale, Francesca Capasso, Alessandra Camarca, Maria V. Barone, Annamaria Staiano,et al.
Oxford University Press (OUP)
Background:The single-nucleotide polymorphism T300A of ATG16L1, a Crohn's disease (CD)–associated gene, is responsible for decreased autophagy. This study aimed to investigate the effects of this single-nucleotide polymorphism on the uptake and processing of antigens by dendritic cells (DCs) and the interaction between DC and intestinal epithelium in pediatric patients with CD. Methods:Pediatric patients who homozygously carry either the protective (wild type, n = 7) or risk allele (risk, n = 13) of ATG16L1, as well as heterozygous patients (het, n = 13) were enrolled. The monocyte-derived DC were analyzed for phenotype, antigen sampling, and processing by flow cytometry, whereas the capability of DC to form transepithelial protrusions was determined by confocal microscopy. Results:DC generated from wild type patients showed higher bacteria sampling and antigen processing compared with risk patients. Additionally, after exposure to either bacteria particles or the antigen DQ-ovalbumin, wild type DC showed a significant increase in the expression of the HLA-DR and CD86 when compared with risk DC. Interestingly, also het patients showed an impairment in bacteria uptake and expression of activation marker when compared with the wild type. In the Caco2/DC coculture, the formation of transepithelial protrusions were less numerous in risk DC compared with wild type and the antigen uptake decreased. Conclusions:DC of pediatric patients with CD carrying the T300A allele showed a marked impairment of antigen uptake and processing and defective interactions between DC and intestinal epithelium. Collectively, our results suggest that an autophagy defect is associated with an impairment of intestinal innate immunity in pediatric CD.
Caterina Strisciuglio, Eleonora Giannetti, Francesca Paola Giugliano, Luigi Greco, Severo Campione, Mariarosaria D’ Armiento, Annamaria Staiano, and Erasmo Miele
Oxford University Press (OUP)
Background: An involvement of the appendiceal orifice as a distintive skip lesion in adults with left side ulcerative colitis (UC) has been reported. The aim of our prospective study was to evaluate, by endoscopy and histology, the prevalence of periappendiceal inflammation (PAI) in children affected by UC. Methods: Fifty of 77 consecutive children undergoing total colonoscopy, who had a diagnosis of UC not extended beyond the hepatic flexure were enrolled. Results: PAI was endoscopically present in 16 of 50 patients (32%) with UC. Patients were divided in 2 groups: group A included the 16 patients with PAI, whereas group B included 34 patients without PAI. We found that among the 2 groups, PAI was more frequent in patients with new diagnosis than in those with pre-existing UC (P = 0.016). At index colonoscopy, the patients of group A had a significant major extent of disease (P = 0.013). Moreover, the histologic grade of inflammation at the ascending colon was significantly higher in group A than in group B (P = 0.014). Clinical activity, measured by pediatric ulcerative colitis activity index, and use of medication did not show significant differences among groups (P = 0.464 and P = 0.723, respectively). The use of immunosuppressant was significantly higher in group A than in group B. Conclusions: PAI is a frequent skip lesion in children with UC. It seems more frequent in patients with new diagnosis, and it is associated with a major extent of the disease and with a higher grade of histologic inflammation at the ascending colon.