Lars Börjesson

The risk of dysplasia and cancer in the ileal pouch mucosa after restorative proctocolectomy for ulcerative proctocolitis is low: a long-term term follow-up study.

Aim  Some of the rare complications reported in patients with an ileopouch anal anastomosis (IPAA) after coloectomy for chronic ulcerative colitis are dysplasia and carcinoma. The supposed pathway is for the ileal pouch mucosa to go through adaptational changes then is to progress through the phases of chronic pouchitis, dysplasia and subsequently to adenocarcinoma. In many of these studies however, the dysplasia – cancer sequence is inconclusive since the carcinoma might have developed from the ileal mucosa itself or from residual viable rectal mucosa left behind. The purpose of this study was therefore to study the long‐term ileal mucosal adaptation patterns and the incidence and grading of dysplasia in the ileal pouch mucosa in patients previously operated on for ulcerative proctocolitis.

Accumulation of CCR4+ CTLA-4hi FOXP3+CD25hi Regulatory T Cells in Colon Adenocarcinomas Correlate to Reduced Activation of Conventional T Cells

Background Colorectal cancer usually gives rise to a specific anti-tumor immune response, but for unknown reasons the resulting immunity is not able to clear the tumor. Recruitment of activated effector lymphocytes to the tumor is important for efficient anti-tumor responses, while the presence of regulatory T cells (Treg) down-modulate tumor-specific immunity. We therefore aimed to determine homing mechanisms and activation stage of Treg and effector T cell infiltrating colon tumors compared to cells from the unaffected mucosa in patients suffering from colon adenocarcinoma. Methodology/Principal Findings Lymphocytes were isolated from unaffected and tumor mucosa from patients with colon adenocarcinoma, and flow cytometry, immunohistochemistry, and quantitative PCR was used to investigate the homing mechanisms and activation stage of infiltrating Treg and conventional lymphocytes. We detected significantly higher frequencies of CD25highFOXP3+CD127low putative Treg in tumors than unaffected mucosa, which had a complete demethylation in the FOXP3 promotor. Tumor-associated Treg had a high expression of CTLA-4, and some appeared to be antigen experienced effector/memory cells based on their expression of αEβ7 (CD103). There were also significantly fewer activated T cells and more CTLA-4+ conventional T cells susceptible to immune regulation in the tumor-associated mucosa. In contrast, CD8+granzyme B+ putative cytotoxic cells were efficiently recruited to the tumors. The frequencies of cells expressing α4β7 and the Th1 associated chemokine receptor CXCR3 were significantly decreased among CD4+ T cells in the tumor, while frequencies of CD4+CCR4+ lymphocytes were significantly increased. Conclusions/Significance This study shows that CCR4+CTLA4hi Treg accumulate in colon tumors, while the frequencies of activated conventional Th1 type T cells are decreased. The altered lymphocyte composition in colon tumors will probably diminish the ability of the immune system to effectively attack tumor cells, and reducing the Treg activity is an important challenge for future immunotherapy protocols.

DMPP causes relaxation of rat distal colon by a purinergic and a nitrergic mechanism

The non-adrenergic relaxation of carbachol precontracted longitudinal muscle of the rat distal colon was investigated. Intrinsic nerves were activated by the nicotinic, ganglionic receptor agonist, 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP). DMPP at 1 and 4 microM caused a relaxation that was markedly antagonized by the nerve blocker tetrodotoxin (1 microM) or the nicotinic receptor antagonist, hexamethonium (1 mM). The response to DMPP was significantly antagonized by apamin (an inhibitor of ATP-sensitive K+-channels), by reactive blue 2 (a blocker of P2y purinoceptors) and by an inhibitor of nitric oxide (NO)-synthase (N(G)-nitro-L-arginine, L-NNA). The combined treatment with reactive blue 2 and L-NNA reduced the relaxatory response to 1 microM DMPP by 77 +/- 8% and to 4 microM DMPP by 58 +/- 4% of control, but left a residual component. Our results indicate that ATP and NO, together with at least one additional (hitherto unidentified) substance may be inhibitory neurotransmitters in rat distal colon.

Macrophage and dendritic cell subsets in IBD: ALDH + cells are reduced in colon tissue of patients with ulcerative colitis regardless of inflammation

Disruption of the homeostatic balance of intestinal dendritic cells (DCs) and macrophages (MQs) may contribute to inflammatory bowel disease. We characterized DC and MQ populations, including their ability to produce retinoic acid, in clinical material encompassing Crohn’s ileitis, Crohn’s colitis and ulcerative colitis (UC) as well as mesenteric lymph nodes (MLNs) draining these sites. Increased CD14+DRint MQs characterized inflamed intestinal mucosa while total CD141+ or CD1c+ DCs numbers were unchanged. However, CD103+ DCs, including CD141+CD103+ and CD1c+CD103+ DCs, were reduced in inflamed intestine. In MLNs, two CD14− DC populations were identified: CD11cintHLADRhi and CD11chiHLADRint cells. A marked increase of CD11chiHLADRint DC, particularly DRintCD1c+ DCs, characterized MLNs draining inflamed intestine. The fraction of DC and MQ populations expressing aldehyde dehydrogenase (ALDH) activity, reflecting retinoic acid synthesis, in UC colon, both in active disease and remission, were reduced compared to controls and inflamed Crohn’s colon. In contrast, no difference in the frequency of ALDH+ cells among blood precursors was detected between UC patients in remission and non-inflamed controls. This suggests that ALDH activity in myeloid cells in the colon of UC patients, regardless of whether the disease is active or in remission, is influenced by the intestinal environment.

Human Mucosa-Associated Invariant T Cells Accumulate in Colon Adenocarcinomas but Produce Reduced Amounts of IFN-γ

Mucosa-associated invariant T (MAIT) cells are innate-like T cells with a conserved TCR α-chain recognizing bacterial metabolites presented on the invariant MHC-related 1 molecule. MAIT cells are present in intestinal tissues and liver, and they rapidly secrete IFN-γ and IL-17 in response to bacterial insult. In colon cancer, IL-17–driven inflammation promotes tumor progression, whereas IFN-γ production is essential for antitumor immunity. Thus, tumor-associated MAIT cells may affect antitumor immune responses by their secreted cytokines. However, the knowledge of MAIT cell presence and function in tumors is virtually absent. In this study, we determined the frequency, phenotype, and functional capacity of MAIT cells in colon adenocarcinomas and unaffected colon lamina propria. Flow cytometric analyses showed significant accumulation of MAIT cells in tumor tissue, irrespective of tumor stage or localization. Colonic MAIT cells displayed an activated memory phenotype and expression of chemokine receptors CCR6 and CCR9. Most MAIT cells in unaffected colon tissues produced IFN-γ, whereas only few produced IL-17. Colonic MAIT cells also produced TNF-α, IL-2, and granzyme B. In the tumors, significantly lower frequencies of IFN-γ–producing MAIT cells were seen, whereas there were no differences in the other cytokines analyzed, and in vitro studies showed that secreted factors from tumor tissue reduced IFN-γ production from MAIT cells. In conclusion, MAIT cells infiltrate colon tumors but their ability to produce IFN-γ is substantially reduced. We suggest that MAIT cells have the capacity to promote local immune responses to tumors, but factors in the tumor microenvironment act to reduce MAIT cell IFN-γ production.

Increased concentration of tissue-degrading matrix metalloproteinases and their inhibitor in complicated diverticular disease

Objective. Complicated diverticular disease is associated with extensive structural changes of the colonic wall. Turnover of extracellular matrix (ECM) plays a pivotal role in this process. Proteolytic enzymes, including matrix metalloproteinases (MMPs), are capable of degrading most components of ECM. Their activity is regulated by inhibitors, tissue inhibitors of metalloproteinases (TIMPs). Disturbances of the MMP-TIMP balance can cause tissue degradation or fibrosis. The aim of this study was to assess the concentration and distribution of MMPs and TIMPs in colonic biopsies. Material andmethods. Twenty-seven patients who had undergone sigmoid colectomy were included in the study. Full-thickness biopsies from affected and non-affected parts of each resected specimen were collected. Expressions of the proteins MMP-1, -2, -3, -9, TIMP-1 and TIMP-2 were quantified by ELISA and localized by immunohistochemistry. Results. The concentrations of MMP-1, MMP-2 and TIMP-1 were significantly higher in affected tissue than concentrations in non-affected tissue (MMP-1 p=0.005, MMP-2 p=0.0003 and TIMP-1 p<0.0001). In affected segments in general, there was an increased expression in the entire bowel wall, predominantly for MMP-2, MMP-3 and TIMP-1. Conclusions. Concentrations of MMP-1, MMP-2 and TIMP-1 were increased in intestinal segments affected by complicated diverticular disease and distributed throughout the entire bowel wall, which may explain the structural changes.

Long-term function and manovolumetric characteristics after ileal pouch-anal anastomosis for ulcerative colitis.

Long‐term pouch function and physiological characteristics after ileal pouch–anal anastomosis (IPAA) are poorly described. The aim of this study was to undertake a prospective investigation of long‐term pouch function and manovolumetric characteristics.

Erratum: The efficacy of a nerve stimulator (Cavermap®) to enhance autonomic nerve identification and confirm nerve preservation during total mesorectal excision (Diseases of the Colon and Rectum (December 2004) 47:12 (2032-2038) DOI: 10.1007/s10350-004-0718-5)

PURPOSESexual dysfunction after total mesorectal excision may be caused by injury to the autonomic nerves. During surgery, nerve identification is not always achieved, and, to date, there has been no method to objectively confirm nerve preservation. The aim of this study was to assess the efficacy of a nerve-stimulating device (CaverMap®) to assist in the intraoperative identification of the autonomic nerves during total mesorectal excision, and objectively confirm nerve preservation after proctectomy is completed.PATIENTS AND METHODSSexually active consecutive male patients undergoing total mesorectal excision were prospectively enrolled in this study. During pelvic dissection, the surgeon attempted to localize the hypogastric and cavernous nerves. Cavermap® was used to confirm these findings and to facilitate the identification in cases of uncertainty. At the completion of proctectomy, the nerves were restimulated to ensure preservation. Factors that could affect the surgeons ability to localize the nerves and Cavermap® to confirm this were evaluated.RESULTSTwenty-nine male patients with a median age of 58 years were enrolled in this study. An attempt to visualize the hypogastric nerves during dissection was made in 26 patients; the surgeon was able to identify the nerves in 19 (73 percent) patients. Cavermap® successfully identified the nerves in six of the seven remaining patients, and failed to identify them in only one case. An attempt to localize the cavernous nerves during dissection was made in 13 patients, of which localization was successful in 8 (61.5 percent) patients. Cavermap® improved the identification rate in four of the remaining five patients. After proctectomy, Cavermap® successfully confirmed the preservation of both hypogastric and cavernous nerves in 27 of 29 (93 percent) patients. A history of previous surgery statistically correlated with failure to identify the hypogastric nerves by the surgeon (P = 0.005). There were no adverse events related to use of the device.CONCLUSIONCavermap® may be a useful tool to facilitate identification of the pelvic autonomic nerves during total mesorectal excision and to objectively confirm nerve preservation.

Altered expression of Butyrophilin (BTN) and BTN-like (BTNL) genes in intestinal inflammation and colon cancer

Several Butyrophilin (BTN) and Btn‐like (BTNL) molecules control T lymphocyte responses, and are genetically associated with inflammatory disorders and cancer. In this study, we present a comprehensive expression analysis of human and murine BTN and BTNL genes in conditions associated with intestinal inflammation and cancer. Using real‐time PCR, expression of human BTN and BTNL genes was analyzed in samples from patients with ulcerative colitis, irritable bowel syndrome, and colon tumors. Expression of murine Btn and Btnl genes was examined in mouse models of spontaneous colitis (Muc2−/−) and intestinal tumorigenesis (ApcMin/+). Our analysis indicates a strong association of several of the human genes with ulcerative colitis and colon cancer; while especially BTN1A1, BTN2A2, BTN3A3, and BTNL8 were significantly altered in inflammation, colonic tumors exhibited significantly decreased levels of BTNL2, BTNL3, BTNL8, and BTNL9 as compared to unaffected tissue. Colonic inflammation in Muc2−/− mice significantly down‐regulated the expression of particularly Btnl1, Btnl4, and Btnl6 mRNA, and intestinal polyps derived from ApcMin/+ mice displayed altered levels of Btn1a1, Btn2a2, and Btnl1 transcripts. Thus, our data present an association of BTN and BTNL genes with intestinal inflammation and cancer and represent a valuable resource for further studies of this gene family.

Pouch design and long-term functional outcome after ileal pouch-anal anastomosis

Functional outcome is of utmost importance after ileal pouch–anal anastomosis. Although pouch design and construction of the anastomosis are known technical determinants of function, there are few long‐term results. This retrospective study evaluated functional outcome for two different pouch designs, and for handsewn versus stapled pouch–anal anastomoses.