CD163L1+CXCL10+ Macrophages are Enriched Within Colonic Lamina Propria of Diverticulitis Patients.

BACKGROUND: Inflammation of diverticula, which are outpouchings of the colonic bowl wall, causes diverticulitis. Severe cases of diverticulitis require surgical intervention. Through RNA-seq analysis of intestinal tissues, we previously found that the innate immune response was deregulated in surgical diverticulitis patients. In that study, pro-inflammatory and macrophage markers were differentially expressed in the colons of diverticulitis versus control patients. Here we investigate CD163L1+ macrophages and the pro-inflammatory chemokine, CXCL10, in diverticulitis. MATERIALS AND METHODS: We assessed tissue from an uninvolved area adjacent to a region of the sigmoid colon chronically affected by diverticulitis and performed Spearman's correlation on transcripts associated with macrophage signaling. We identified altered CD163L1 and CXCL10 gene expression levels that we confirmed by RT-qPCR analysis on an independent cohort of diverticulitis patients and controls. We used immunofluorescence microscopy to localize CD163L1+ macrophages and CXCL10 levels in intestinal tissue and ELISA to measure CXCL10 levels in patient serum. RESULTS: We found a positive correlation between intestinal CD163L1 and CXCL10 gene expression and an increased number of CD163L1+ macrophages in the sigmoid colons of diverticulitis patients relative to controls (P = 0.036). Macrophages at the apices of colonic crypts expressed the chemokine CXCL10. Correspondingly, these diverticulitis patients also displayed heightened CXCL10 levels in their serum (P = 0.007). CONCLUSIONS: We identified a novel population of CD163L1+CXCL10+ macrophages in the colonic crypts of diverticulitis patients and demonstrated increased expression of serum CXCL10 in these patients. CXCL10 may serve as a prognostic biomarker to aid in clinical decision making for diverticulitis patients.

Diverticulitis en pacientes inmunodeprimidos: resultados de nuestra unidad en el manejo de pacientes de alto riesgo / No disponible

INTRODUCCIÓN: la diverticulitis aguda es una enfermedad con una gran prevalencia y la necesidad de un manejo más agresivo en pacientes inmunodeprimidos no ha quedado claramente consensuada. OBJETIVOS: valorar la posibilidad de que el manejo conservador en este grupo sea tan válido como en la población inmunocompetente. MÉTODOS: presentamos un estudio analítico retrospectivo llevado a cabo en nuestro hospital. Se analizaron 40 pacientes inmunodeprimidos (trasplantados, tratamiento corticoideo, tratamiento renal sustitutivo, oncológicos, VIH positivos en situación de inmunodepresión) diagnosticados de diverticulitis aguda. Los pacientes siguieron manejo no quirúrgico o tratamiento quirúrgico urgente según su situación clínica al ingreso. Las principales medidas estudiadas han sido la gravedad del cuadro agudo y la necesidad de tratamiento quirúrgico según la causa de inmunodepresión. RESULTADOS: de 40 pacientes estudiados, 32 fueron manejados de forma conservadora en el momento agudo, con necesidad de intervención quirúrgica urgente en ocho casos (siete intervenciones de Hartmann y una resección con anastomosis), siendo el grupo de los trasplantados y los pertenecientes al rango de edad 40-50 años los de mayor riesgo. Tres pacientes requirieron intervención de forma programada por complicaciones posteriores. Veinticuatro pacientes evolucionaron sin complicaciones. CONCLUSIONES: la proporción de diverticulitis aguda complicada es superior que en la población general. El tratamiento no quirúrgico parece tan seguro como en la población inmunocompetente. Los pacientes más jóvenes y los trasplantados constituyen los principales grupos de riesgo para sufrir un cuadro grave que precise un manejo más agresivo de manera inicial

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The efficacy of a mix of three probiotic strains in reducing abdominal pain and inflammatory biomarkers in acute uncomplicated diverticulitis.

OBJECTIVE: Acute Uncomplicated Diverticulitis (AUD) is defined as the inflammation of a colon diverticulum, often involving colic wall and pericolic fat. Conventional treatment of AUD includes antibiotics, usually ciprofloxacin and metronidazole, fasting, and fluid therapy. The aim of this study was to test the efficacy of a mix of three probiotic strains (Bifidobacterium lactis LA 304, Lactobacillus salivarius LA 302, Lactobacillus acidophilus LA 201; Lactibiane Iki®, Biocure [PiLeJe Groupe], Italy/PiLeJe Laboratoire, France) in association with conventional antibiotics in treating AUD compared to conventional antibiotics used alone. PATIENTS AND METHODS: We enrolled 84 (25M/59F mean age 61.5 ± 11.5 years) consecutive patients who came to the Emergency Department of the Fondazione Policlinico Universitario A. Gemelli - IRCCS, Rome, Italy, with a diagnosis of AUD confirmed by CT scan. After routine blood test and dosage of C-reactive protein (C-RP), patients were randomly divided into two groups: Probiotic group (42 patients, 10M/32F mean age 32.23 ± 10.3 years) was treated with ciprofloxacin 400 mg twice a day and metronidazole 500 mg three times a day for one week and simultaneously supplemented with the probiotic mix, 1 sachet twice a day for 10 days. Control group (42 patients, 15M/27F mean age 59.01 ± 11.3 years) received the same antibiotic treatment without the probiotic mix. All patients filled a daily Visual Analog Scale (VAS) for assessment of abdominal pain, with a range value from 0 (asymptomatic) to 10, and CRP value was determined on admission and at discharge. RESULTS: As regards abdominal pain, on Day 3, Group A showed a significant decrease of 4.06 points (51.4%) in VAS score compared to a decrease of 2.79 points (34.9%) in Group B. On Day 5 the decrease was of 6.3 points (80%) in Group A and of 4.85 points (61%) in Group B. VAS score was reduced by 7.59 points (96%) in Group A and 6.1 points (76%) in Group B on Day 7 +, and by 7.8 points (99%) in Group A and 7.2 points (90%) in Group B on Day 10. About inflammation, Group A showed a decrease in C-RP value of 64%, compared to a decrease of only 35% in Group B. We also observed that the duration of hospitalization was significantly shorter for patients in Group A: 89 h (3.7 days) in Group A vs. 101 h (4.2 days) in Group B (p=0.03). CONCLUSIONS: Our results indicated showed that the supplement with the probiotic mix of Bifidobacterium lactis LA 304, Lactobacillus salivarius LA 302, and Lactobacillus acidophilus LA 201 in combination with the standard antibiotic therapy for AUD reduced abdominal pain and inflammation significantly more than antibiotic treatment used alone. These findings could be due to the anti-inflammatory activity of the probiotic mix. Larger studies are needed to validate its use in the clinical practice.

RNA-seq implicates deregulation of the immune system in the pathogenesis of diverticulitis.

Individuals with diverticula or outpouchings of the colonic mucosa and submucosa through the colonic wall have diverticulosis, which is usually asymptomatic. In 10-25% of individuals, the diverticula become inflamed, resulting in diverticulitis. Very little is known about the pathophysiology or gene regulatory pathways involved in the development of diverticulitis. To identify these pathways, we deep sequenced RNAs isolated from full-thickness sections of sigmoid colon from diverticulitis patients and control individuals. Specifically for diverticulitis cases, we analyzed tissue adjacent to areas affected by chronic disease. Since the tissue was collected during elective sigmoid resection, the disease was in a quiescent state. A comparison of differentially expressed genes found that gene ontology (GO) pathways associated with the immune response were upregulated in diverticulitis patients compared with nondiverticulosis controls. Next, weighted gene coexpression network analysis was performed to identify the interaction among coexpressed genes. This analysis revealed RASAL3, SASH3, PTPRC, and INPP5D as hub genes within the brown module eigengene, which highly correlated (r = 0.67, P = 0.0004) with diverticulitis. Additionally, we identified elevated expression of downstream interacting genes. In summary, transcripts associated with the immune response were upregulated in adjacent tissue from the sigmoid colons of chronic, recurrent diverticulitis patients. Further elucidating the genetic or epigenetic mechanisms associated with these alterations can help identify those at risk for chronic disease and may assist in clinical decision management.NEW & NOTEWORTHY By using an unbiased approach to analyze transcripts expressed in unaffected colonic tissues adjacent to those affected by chronic diverticulitis, our study implicates that a defect in the immune response may be involved in the development of the disease. This finding expands on the current data that suggest the pathophysiology of diverticulitis is mediated by dietary, age, and obesity-related factors. Further characterizing the immunologic differences in diverticulitis may better inform clinical decision-making.

Diverticulitis in transplant patients and patients on chronic corticosteroid therapy: a systematic review.

BACKGROUND: The clinical course of diverticular disease in immunosuppressed patients is widely believed to be more severe than in the general population. In this study we systematically reviewed the literature regarding the epidemiology and clinical course of diverticulitis in immunosuppressed patients. Our goal was to develop recommendations regarding the care of this group of patients. METHODS: Using PubMed and Web of Knowledge we systematically reviewed all studies published between 1970 and 2009 that analyzed the epidemiology, clinical manifestation, or outcomes of treatment of diverticulitis in immunosuppressed patients. Keywords of "transplantation," "corticosteroid," "HIV," "AIDS," and "chemotherapy" were used. RESULTS: Twenty-five studies met our inclusion criteria. All of these studies focused on the impact of diverticulitis in patients with transplants or on chronic corticosteroid therapy. The reported incidence of acute diverticulitis in these patients was approximately 1% (variable follow-up periods). Among patients with known diverticular disease the incidence was 8%. Mortality from acute diverticulitis in these patients was 23% when treated surgically and 56% when treated medically. Overall mortality was 25%. CONCLUSIONS: Our study summarizes evidence that patients with transplants or patients on chronic corticosteroid therapy 1) have a rate of acute diverticulitis that is higher than the baseline population and 2) a mortality rate with acute diverticulitis that is high. Further research is needed to define whether these risks constitute a mandate for screening and prophylactic sigmoid colectomy.

Post inflammatory damage to the enteric nervous system in diverticular disease and its relationship to symptoms.

Some patients with colonic diverticula suffer recurrent abdominal pain and exhibit visceral hypersensitivity, though the mechanism is unclear. Prior diverticulitis increases the risk of being symptomatic while experimental colitis in animals increases expression of neuropeptides within the enteric nervous system (ENS) which may mediate visceral hypersensitivity. Our aim was to determine the expression of neuropeptides within the ENS in diverticulitis (study 1) and in patients with symptomatic disease (study 2). Study 1 - Nerves in colonic resection specimens with either acute diverticulitis (AD, n = 16) or chronic diverticulitis (CD, n = 16) were assessed for neuropeptide expression recording % area staining with protein gene product (PGP9.5), substance P (SP), neuropeptide K (NPK), pituitary adenylate cyclase activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP) and galanin. Study 2 - Seventeen symptomatic and 15 asymptomatic patients with colonic diverticula underwent flexible sigmoidoscopy and multiple peridiverticular mucosal biopsies. Study 1- Neural tissue, as assessed by PGP staining was increased to a similar degree in circular muscle in both AD and CD. The CD specimens showed significant increases in the immunoreactivity of SP, NPK and galanin in both mucosal and circular muscle layer compared with controls. Study 2 - Mucosal histology was normal and PGP9.5 staining was similar between groups however patients with symptomatic diverticular disease demonstrated significantly higher levels of SP, NPK, VIP, PACAP and galanin within the mucosal plexus. Patients with symptomatic diverticular disease exhibit increased neuropeptides in mucosal biopsies which may reflect resolved prior inflammation, as it parallels the changes seen in acute and chronic diverticulitis.

Acute recurrent diverticulitis is prevented by oral administration of a polybacterial lysate suspension.

AIM: The main cause of acute diverticulitis is the abnormal accumulation of fecal bacteria within the diverticular lumen, leading to a balancing between normal probiotic microflora and pathogenic species; Gram negative Entero-bacteriaceae, mainly Escherichia coli and Proteus spp, are the genders that usually cause the disease-related symptoms, due to their ability to adhere to intestinal mucosa. The intestine is well known as the largest human lymphoepithelial organ and daily produces more antibodies, mainly secretory IgAs, than do all other lymphoid tissues. IgAs have different immune and anti-inflammatory properties. The aim of this study was to verify the efficacy of an oral immunostimulant highly-purified, polymicrobial lysate in the prevention of recurrent attacks of diverticulitis and in the improvement of symptoms. METHODS: The study was carried out on 83 consecutive patients suffering from recurrent symptomatic acute diverticulitis and with at least 2 attacks in the previous year; patients were randomly assigned to receive (group A) an oral polybacterial lysate suspension or to a no-treatment clinical follow-up as controls (group B). RESULTS: A total of 76 patients (41 in group A and 35 in group B) terminated the study period. the sums of the scores for symptoms, reported on day schedules, were calculated and examined by means of ANOVA statistical analysis. Statistical differences between group A vs group B were recorded after 1 month (p<0.05) and 3 months (p<0.01) of treatment with the oral polybacterial lysate suspension. CONCLUSIONS: Our data suggest that the administration of an oral enterovaccine for the prophylaxis of recurrent diverticulitis is effective and well tolerated, probably due to a direct stimulation of IgA-mediated mucosal defences.

The severity of infection induces a shift in the type 1/Type 2 T-helper cell balance in patients with or without peritonitis.

BACKGROUND: A type-1 to type-2 T-helper cell (Th1/Th2) shift is hypothesized to occur among patients with severe trauma and pancreatitis. It was hypothesized that the Th2 situation caused an immune-suppressed period that led to a critical imbalance in the patients' conditions, with an increased risk of multiple organ failure and mortality. METHODS: In eight patients with localized intra-abdominal infection (diverticulitis) and six patients with peritonitis due to perforation of a hollow viscus, we examined the cytokine response of CD3(+) T cells in the greater omentum and in the peripheral blood by in vitro stimulation, intracellular cytokine staining, and flow cytometry for TNF-alpha, IFN-gamma, IL-2, and IL-4. Follow-up cytokine assays were carried out on peripheral blood on days 3 and 7. Different levels of cytokine expression in each group were examined to determine the origin of the lymphocytes, both from omentum or peripheral blood. Cytokine production in the diverticulitis group was compared with that of the peritonitis patients. RESULTS: In localized infections (diverticulitis), there was higher expression of TNF-alpha (51%/35% positive cells in omentum/blood), IFN-gamma (47%/32%), and IL-2 (33%/20%) in the omentum than in the peripheral blood mononuclear cells (PMBCs), but this was not true for IL-4 expression (0.8%/1.3%). In patients with peritonitis, there were no differences in cytokine expression between lymphocytes from the greater omentum and from PMBCs for TNF-alpha (18%/21% omentum/blood), IFN-gamma (20%/22%), IL-2 (16%/12%), or IL-4 (10.9%/7.6%). Compared to the diverticulitis group, patients with peritonitis showed reduced expression for TNF-alpha, IFN-gamma, and IL-2, but there was a significantly higher response for IL-4 for both compartments. CONCLUSIONS: There was a shift from Th1 to Th2 in patients with severe clinical symptoms of peritonitis. Immune suppression is evident because of the T cell response in the greater omentum, but immunosuppression seems to not reach its maximum level before day 7 post operation. This differs from findings in multiple trauma and pancreatitis; however, it is parallel to the clinical situation in patients with peritonitis.

Lamina propria T cells in Crohn's disease and other gastrointestinal inflammation show defective CD2 pathway-induced apoptosis.

BACKGROUND & AIMS: Normal human lamina propria lymphocytes manifest increased unstimulated apoptosis compared with peripheral lymphocytes, which are enhanced after stimulation via the CD2 activation pathway. This activation-induced apoptosis down-regulates cell expansion and cytokine production. In previous studies, it was shown that lamina propria T cells from patients with Crohn's disease and ulcerative colitis manifest abnormal proliferation and cytokine production. It was therefore of interest to determine if such cells also showed abnormal patterns of apoptosis. METHODS: Apoptosis was evaluated by propidium iodide staining of cells followed by flow cytometric analysis. Fas expression and Bcl-2 levels in cells were evaluated by immunofluorescence. RESULTS: Lamina propria lymphocytes from patients with Crohn's disease and ulcerative colitis as well as from 2 patients with diverticulitis showed defective CD2 pathway-induced apoptosis. Studies of the mechanisms of this defect focusing on cells from patients with Crohn's disease showed that Crohn's disease lamina propria lymphocytes from inflamed tissues express the same amount of cell surface Fas but are less sensitive to Fas-mediated apoptosis than control cells. In addition, lamina propria lymphocytes from inflamed Crohn's disease tissues manifest increased expression of Bcl-2 after CD2 pathway stimulation and elevated Bcl-2 levels in cultures of unstimulated T cells. CONCLUSIONS: T cells isolated from areas of inflammation in Crohn's disease, ulcerative colitis, and other inflammatory states manifest decreased CD2 pathway-induced apoptosis. Studies of cells from inflamed Crohn's disease tissue indicate that this defect is accompanied by elevated Bcl-2 levels. These changes are probably caused by the chronic inflammation and may aggravate the underlying disease processes that are present.

Persistent clonal expansions of peripheral blood CD4+ lymphocytes in chronic inflammatory bowel disease.

It is increasingly recognized that chronic Ag exposure may lead to clonal expansions of T cells, including those within the peripheral blood. Inflammatory bowel disease is a chronic, multisystemic disease of unknown origin that predominantly affects the intestine. We sought to determine whether clonal expansions of T cells are present in the peripheral blood of patients with inflammatory bowel disease by an examination of TCR usage. Positively selected CD4+ and CD8+ peripheral blood T cells were isolated from subjects with active ulcerative colitis, Crohn's disease, and diverticulitis and from normal controls. Analysis of complementarity determining region 3 lengths of 24 TCR-beta chain V region families from CD4+ and CD8+ peripheral blood T cells showed a skewed distribution in the three inflammatory groups, consistent with expansion of T cell clones, in comparison to the normally distributed pattern observed among the control donors. Random sequencing of the PCR amplification products of CD4+ peripheral blood T cells from the subjects with ulcerative colitis, Crohn's disease, and diverticulitis revealed reiterative TCR-beta chain sequences that were not found in the normal donors. In subjects with Crohn's disease, the reiterative TCR-beta chain sequences from the CD4+ peripheral blood T cells were persistent over at least a 1-yr period. The persistently expanded TCR-beta chain sequences of CD4+ peripheral blood T cells were identifiable in genomic DNA isolated from archival tissue of intestine from subjects with Crohn's disease and ulcerative colitis by Southern blotting and direct DNA sequencing. An identical twin pair, concordant for Crohn's disease, shared the same reiterative TCR-beta chain sequences in their CD4+ peripheral blood T cells. These studies show that chronic intestinal inflammation is associated with expansions of CD4+ peripheral blood T cells. Furthermore, in inflammatory bowel disease these T cell clonal expansions are persistent and shared among HLA-identical individuals, implicating a response to specific, persistent, and stimulating Ags in these diseases.

Over-utilization of the J delta 3 gene-segment in Crohn's disease.

A majority of normal human intestinal intraepithelial lymphocytes (iIEL) are CD8+, express the alpha beta-T cell receptor (TCR) and are oligoclonal. The remainder of normal iIELs, which are also oligoclonal, express the gamma delta-TCR and preferentially utilize variable regions (V delta 1 and V delta 3) which are different from adult peripheral blood lymphocytes (V delta 2). The junctional region usage of gamma delta-TCRs in intestinal diseases is largely unknown. The aim of this study was to examine gamma delta-T cell clonality and junctional region usage of V delta 1 and V delta 3 transcripts in Crohn's Disease (CD) in comparison to several other chronic inflammatory diseases of the colon by polymerase chain reaction amplification, cloning and sequencing. As previously observed in normal subjects, all inflammatory cases examined, including CD (n = 3), ulcerative colitis (n = 1), diverticulitis (n = 1) and lymphocytic colitis (n = 1), the V delta 1 and V delta 3 transcripts contained reiterated sequences consistent with the expansion of gamma delta-T cells expressing these receptors. In 2/3 CD cases, but none of the non-CD inflammatory cases, transcripts containing J delta 3, a rarely used J delta, was observed among the V delta 1 and/or V delta 3 transcripts. Thus, in a subset of CD, gamma delta-T cells expressing J delta 3 may be expanded implicating a role for unique ligands that drive the expansion of T cells expressing these receptors.

Gland to gland heterogeneity in histologically normal mucosa of colon cancer patients demonstrated by monoclonal antibodies to tissue-specific antigens.

Two of three monoclonal antibodies to tissue-specific antigens of isolated colonic glands revealed gland to gland heterogeneity of antigen expression in sections of the histologically normal colonic mucosa from patients with colorectal adenocarcinoma or various nonmalignant conditions. A colon-specific goblet cell antigen (designated 3NM) was absent from rare, solitary glands randomly distributed among the otherwise strongly stained glands of distal colon. These 3NM-negative glands stained normally for the other two antigens studied and appeared morphologically and histochemically normal. They occurred in 11 of 13 cancer patients and in each of the five patients with benign conditions with median incidences of 2.2 and 0.4 per 1000 glands, respectively. Gland heterogeneity was also demonstrated in both patient groups for a cell membrane antigen (designated 6NM). In cecum and ascending colon, glands staining strongly and weakly for 6NM were found intermixed. The weakly stained glands tended to predominate in cecum and first part of ascending colon, but they were completely replaced by strongly stained glands in more distal colon. The heterogeneity shown by both 3NM and 6NM appeared due to phenotypically distinct cell clones. Our observations indicate that histologically normal colonic mucosa contains antigenically diverse gland populations.

Appearance of dextrans and antidextran antibodies in human sera.

The serologically active moiety of an antigen detected occasionally in pathologic sera by double-diffusion gel precipitation tests, referred to in earlier studies as ubiquitous tissue antigen, was identified as a dextran composed predominantly, if not exclusively, of a(1 leads to 6)-linked glycopyranoses. By means of an enzyme immunoassay, dextran or dextran-like material, which inhibited the binding of antidextran serum to dextran, was detected in sera of several patients with various gastrointestinal (GI) diseases, especially GI ulcers (7/10), and also often in sera of aged people (9/21). However, 2 of 50 normal blood donor sera and a few sera from almost every disease group studied contained low quantities of dextran-like material. The levels of antidextran antibodies of the IgG class were also often elevated among patients with GI diseases and aged people as demonstrated by enzyme immunoassay with dextran T-500 as the solid phase antigen. OD values exceeding the mean plus 2.5 SD of 106 normal blood donor sera were recorded in ;69% of patients with gastric and duodenal ulcers, 40% with ulcerative colitis, 29% with Crohn's disease, 20% with colorectal carcinomas, and in 21% with rheumatoid arthritis. None of 23 children, but 9 of 23 aged people (35%) had elevated antibody levels. It is suggested that absorption of dextrans from food or their production by intestinal bacteria may be facilitated in various GI diseases.