Biliary Tract Cancer: Implicated Immune-Mediated Pathways and Their Associated Potential Targets.

There is a well-established link between biliary tract cancers (BTC) and chronic inflammatory conditions such as primary sclerosing cholangitis, chronic cholecystitis, chronic cholelithiasis, liver fluke-associated infestations, and chronic viral hepatic infections. These associated risk factors highlight the potential for development of immune-modulatory agents in this poor-prognostic disease group with limited treatment options. Clinical trials have evaluated the role of immune cells, inflammatory biomarkers, vaccines, cytokines, adoptive cell therapy, and immune checkpoint inhibitors in patients with BTC. Although these have demonstrated the importance of the immune environment in BTC, currently none of the immune-based therapies have been approved for use in this disease group. The role of immunomodulatory agents is a developing field and has yet to find its way 'from bench to bedside' in BTC.

COMPARATIVE ANALYSIS OF IMMUNOLOGICAL PROFILES IN WOMEN UNDERGOING CONVENTIONAL AND SINGLE-PORT LAPAROSCOPIC CHOLECYSTECTOMY.

Background: Surgical trauma triggers an important postoperative stress response characterized by significantly elevated levels of cytokines, an event that can favor the emergence of immune disorders which lead to disturbances in the patient's body defense. The magnitude of postoperative stress is related to the degree of surgical trauma. Aim: To evaluate the expression of pro-inflammatory (TNF-α, IFN-γ, IL-1ß, and IL-17) and anti-inflammatory (IL-4) cytokines in patients submitted to conventional and single-port laparoscopic cholecystectomy before and 24 h after surgery. Methods: Forty women with symptomatic cholelithiasis, ranging in age from 18 to 70 years, participated in the study. The patients were divided into two groups: 21 submitted to conventional laparoscopic cholecystectomy and 19 to single-port laparoscopic cholecystectomy. Results: Evaluation of the immune response showed no significant difference in IFN-γ and IL-1ß levels between the groups or time points analyzed. With respect to TNF-α and IL-4, serum levels below the detection limit (10 pg/ml) were observed in the two groups and at the time points analyzed. Significantly higher postoperative expression of IL-17A was detected in patients submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels (p=0.0094). Conclusions: Significant postoperative expression of IL-17 was observed in the group submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels, indicating that surgical stress in this group was higher compared to the conventional laparoscopic cholecystectomy.

Racional: O trauma cirúrgico induz resposta de estresse pós-operatório significativo, evidenciado pelos níveis elevados de citocinas, podendo favorecer o surgimento de distúrbios imunológicos. A magnitude de estresse está relacionada ao grau do trauma cirúrgico. Objetivos: Avaliar a expressão das citocinas pró-inflamatórias (TNF-α, IFN-γ, IL-1ß, IL-17) e da anti-inflamatória (IL-4) no pré e pós-operatório de pacientes submetidas à colecistectomia laparoscópica por dois métodos: convencional e por portal único. Métodos: Quarenta mulheres com colecistolitíase sintomática foram operadas, sendo 21 por procedimento laparoscópico convencional e 19 por portal único. As citocinas TNF-α, IFN-γ, IL-1ß, IL-4, e IL-17 presentes no plasma foram quantificadas pelo método de ELISA em dois momentos: no pré-operatório e após 24 h da operação. Resultados: A avaliação da resposta imune não mostrou diferença estatisticamente significante das citocinas IFN-γ e IL-1ß na comparação entre os grupos e tempos analisados. Em relação às citocinas TNF-α e IL-4 os níveis séricos estavam abaixo dos níveis de detecção (10 pg/ml) em ambos os grupos e tempos analisados. Observou-se que as operadas por portal único apresentaram expressão significativa da IL-17A do período pré para o pós-operatório (p=0,0094). Conclusões: A expressão aumentada da IL-17A no pós-operatório do grupo de portal único pode indicar que o estresse cirúrgico foi maior em comparação ao da colecistectomia laparoscópica convencional.

Comparative analysis of immunological profiles in women undergoing conventional and single-port laparoscopic cholecystectomy / Perfil imunológico de mulheres submetidas à colecistectomia laparoscópica convencional e por portal único

ABSTRACT Background: Surgical trauma triggers an important postoperative stress response characterized by significantly elevated levels of cytokines, an event that can favor the emergence of immune disorders which lead to disturbances in the patient's body defense. The magnitude of postoperative stress is related to the degree of surgical trauma. Aim: To evaluate the expression of pro-inflammatory (TNF-α, IFN-γ, IL-1β, and IL-17) and anti-inflammatory (IL-4) cytokines in patients submitted to conventional and single-port laparoscopic cholecystectomy before and 24 h after surgery. Methods: Forty women with symptomatic cholelithiasis, ranging in age from 18 to 70 years, participated in the study. The patients were divided into two groups: 21 submitted to conventional laparoscopic cholecystectomy and 19 to single-port laparoscopic cholecystectomy. Results: Evaluation of the immune response showed no significant difference in IFN-γ and IL-1β levels between the groups or time points analyzed. With respect to TNF-α and IL-4, serum levels below the detection limit (10 pg/ml) were observed in the two groups and at the time points analyzed. Significantly higher postoperative expression of IL-17A was detected in patients submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels (p=0.0094). Conclusions: Significant postoperative expression of IL-17 was observed in the group submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels, indicating that surgical stress in this group was higher compared to the conventional laparoscopic cholecystectomy.

RESUMO Racional: O trauma cirúrgico induz resposta de estresse pós-operatório significativo, evidenciado pelos níveis elevados de citocinas, podendo favorecer o surgimento de distúrbios imunológicos. A magnitude de estresse está relacionada ao grau do trauma cirúrgico. Objetivos: Avaliar a expressão das citocinas pró-inflamatórias (TNF-α, IFN-γ, IL-1β, IL-17) e da anti-inflamatória (IL-4) no pré e pós-operatório de pacientes submetidas à colecistectomia laparoscópica por dois métodos: convencional e por portal único. Métodos: Quarenta mulheres com colecistolitíase sintomática foram operadas, sendo 21 por procedimento laparoscópico convencional e 19 por portal único. As citocinas TNF-α, IFN-γ, IL-1β, IL-4, e IL-17 presentes no plasma foram quantificadas pelo método de ELISA em dois momentos: no pré-operatório e após 24 h da operação. Resultados: A avaliação da resposta imune não mostrou diferença estatisticamente significante das citocinas IFN-γ e IL-1β na comparação entre os grupos e tempos analisados. Em relação às citocinas TNF-α e IL-4 os níveis séricos estavam abaixo dos níveis de detecção (10 pg/ml) em ambos os grupos e tempos analisados. Observou-se que as operadas por portal único apresentaram expressão significativa da IL-17A do período pré para o pós-operatório (p=0,0094). Conclusões: A expressão aumentada da IL-17A no pós-operatório do grupo de portal único pode indicar que o estresse cirúrgico foi maior em comparação ao da colecistectomia laparoscópica convencional.

Inmunoexpresión p53, sangre oculta en heces de pacientes colecistectomizados y colelitiasis con adenomas colónicos / P53 Immunoexpression, Faecal Occult Blood from Cholecystectomized Patients and Cholelithiasis with Colonic Adenomas

Introducción: la secuencia adenoma- adenocarcinoma, es resultado de fallos genéticos en las células intestinales heredados o adquiridos.Objetivo: determinar la posible relación entre la inmunoexpresión de la p53 y la positividad de la sangre oculta en heces en los adenomas de colon con alto grado de displasia diagnosticados en pacientes colecistectomizados o con colelitiasis.Métodos: se realizó un estudio descriptivo, de corte transversal, en el Instituto de Gastroenterología, en el período de mayo de 2013 hasta junio de 2015. Se realizaron pruebas estadísticas descriptiva y de chi Cuadrado y probabilidad exacta de Fisher.Resultados: la proporción de adenomas con alto grado de displasia fue similar en pacientes colecistectomizados y con colelitiasis (50 por ciento) respectivamente. Una alta proporción se diagnosticó en colecistectomizados femeninas (35 por ciento), con 60 y más años de edad (53 por ciento) y 11 y más años de colecistectomizados (60 por ciento), mientras que en las colelitiasis fueron masculinos (30 por ciento).Conclusiones: una alta proporción de adenomas con alto grado de displasia presentan inmunoexpresión de la p53 y sangre en heces positiva en pacientes colecistectomizados y con colelitiasis, que se reporta por vez primera(AU)

Introduction: The adenoma - adenocarcinoma sequence is a result of inherited or acquired genetic failures in the intestinal cellsObjective: To determine the immunohistochemical expression of p53 and the positivity of the fecal occult blood test in colon adenomas with high degree of diagnosed dysplasia in cholecystectomized patients or with cholelithiasis.Methods: Descriptive, cross-sectional study conducted in the Institute of Gastroenterology in the period of May, 2013 to June, 2015. Statistical tests were statistics testing, exact Chi Square and Fisher's probability tests.Results: The proportion of adenomas with high degree of dysplasia was similar in cholecystectomized patientsand with cholelithiasis (50 percent) respectively. A high proportion diagnosed in colecistectomizados women (35 percent), 60 and more years of age (53 percent) and 11 and more years of performed cholecystectomy (60 percent), whereas cholelithiasis prevailed in males (30 percent).Conclusions: High proportion of adenomas with high degree of dysplasia present p 53 immunoexpression and positive fecal occult blood test in cholecystectomized patients and patients with cholelithiasis that is reported for the first time(AU)

Inmunoexpresión p53, sangre oculta en heces de pacientes colecistectomizados y colelitiasis con adenomas colónicos / P53 Immunoexpression, Faecal Occult Blood from Cholecystectomized Patients and Cholelithiasis with Colonic Adenomas

Introducción: la secuencia adenoma- adenocarcinoma, es resultado de fallos genéticos en las células intestinales heredados o adquiridos. Objetivo: determinar la posible relación entre la inmunoexpresión de la p53 y la positividad de la sangre oculta en heces en los adenomas de colon con alto grado de displasia diagnosticados en pacientes colecistectomizados o con colelitiasis. Métodos: se realizó un estudio descriptivo, de corte transversal, en el Instituto de Gastroenterología, en el período de mayo de 2013 hasta junio de 2015. Se realizaron pruebas estadísticas descriptiva y de chi Cuadrado y probabilidad exacta de Fisher. Resultados: la proporción de adenomas con alto grado de displasia fue similar en pacientes colecistectomizados y con colelitiasis (50 por ciento) respectivamente. Una alta proporción se diagnosticó en colecistectomizados femeninas (35 por ciento), con 60 y más años de edad (53 por ciento) y 11 y más años de colecistectomizados (60 por ciento), mientras que en las colelitiasis fueron masculinos (30 por ciento). Conclusiones: una alta proporción de adenomas con alto grado de displasia presentan inmunoexpresión de la p53 y sangre en heces positiva en pacientes colecistectomizados y con colelitiasis, que se reporta por vez primera(AU)

Introduction: The adenoma - adenocarcinoma sequence is a result of inherited or acquired genetic failures in the intestinal cells. Objective: To determine the immunohistochemical expression of p53 and the positivity of the fecal occult blood test in colon adenomas with high degree of diagnosed dysplasia in cholecystectomized patients or with cholelithiasis. Methods: Descriptive, cross-sectional study conducted in the Institute of Gastroenterology in the period of May, 2013 to June, 2015. Statistical tests were statistics testing, exact Chi Square and Fisher's probability tests. Results: The proportion of adenomas with high degree of dysplasia was similar in cholecystectomized patientsand with cholelithiasis (50 percent) respectively. A high proportion diagnosed in colecistectomizados women (35 percent), 60 and more years of age (53 percent) and 11 and more years of performed cholecystectomy (60 percent), whereas cholelithiasis prevailed in males (30 percent). Conclusions: High proportion of adenomas with high degree of dysplasia present p 53 immunoexpression and positive fecal occult blood test in cholecystectomized patients and patients with cholelithiasis that is reported for the first time(AU)

Laparoscopic cholecystectomy in elderly patients: an evaluation of immunity.

OBJECTIVE: We sought to investigate the impact of laparoscopic cholecystectomy (LC) on the inflammatory response and immunological function of elderly patients compared with that on the younger ones. METHODS: Between June 2012 and June 2013, this prospective study investigated a total of 112 patients having the surgery of LC due to symptomatic cholelithiasis or polyps, among whom 52 were elderly patients with the age beyond 65 years old and the remaining 60 were younger than the age. Peripheral venous blood samples were taken from these patients prior to surgery and on post-operative days 1, 3 and 7, respectively. The perioperative clinical outcomes and immunological function results were analyzed and compared between the two groups divided by age. RESULTS: The demographics of the two groups did not differ except for the age. Surgical trauma seemed more serious for elderly patients as illustrated by the longer operating time, hospital stay and more quantity of patients got complication. Both groups indicated changes in inflammatory and immune aspects. Compared with the younger ones, elderly patients showed less quantity of preoperative basic immune cells, delayed immune responses after the surgical trauma of LC and hyporeactivity of inflammatory response when accepting LC. CONCLUSIONS: An examination of the inflammatory reaction and immune response after LC demonstrated that there are significant differences observed in two groups divided by age. Further studies with more samples are required to determine the exact relationship of perioperative immune change and higher adverse outcome rate of aged people.

[Evaluation of prognostic factors for the development of cholelithiasis in patients with metabolic syndrome].

UNLABELLED: The aim of study to determine the leading clinical and immunological parameters, reflecting the high risk of development and progression of gallstone disease in patients with metabolic syndrome. MATERIALS AND METHODS: An assessment of clinical, biochemical and immunological parameters in 54 patients with gallstone disease associated with the metabolic syndrome and in comparison groups (31 with metabolic syndrome without gallstone disease and 29 with gallstone disease only) were made. For modeling the significant correlation and prediction of the effect of different combinations of factors on the risk of gallstone disease formation in patients with metabolic syndrome was used the multiple logistic regression analysis. RESULTS: In accordance with our results the main risk factors of gallstone formation in patients with metabolic syndrome are the age (P = 0.02), waist volume (P = 0.0002), the increase of serum concentration of GGTP (P = 0.00001), MMP-9 (P = 0.002) and TIMP-1 (P = 0.02). CONCLUSION: The results of our study have shown that in patients with metabolic syndrome was estimated the high risk of gallstone disease formation compared and the most significant factors of it formation are the patients age, waist volume and increase of serum concentration of GGTP, MMP-9 and TIMP-1.

α-Defensins and hsCRP levels in inflammatory response of standard and laparoendoscopic single-site cholecystectomy.

BACKGROUND: Laparoendoscopic single-site (LESS) surgery is an evolution of laparoscopic surgery aiming at decreasing the patient's parietal trauma associated with abdominal operations. LESS has been found so far to be efficient and have the same good results as the standard four-port laparoscopic cholecystectomy. α-Defensins are antimicrobial peptides of the organism. They are the first cell components against pathogens. Cytokines are also mediators in the response to trauma. The aim of this study was to compare the inflammatory reaction in LESS and four-port laparoscopic cholecystectomy. METHODS: Forty patients with noncomplicated cholelithiasis were randomly assigned into one of two groups. Group A included the patients who would undergo four-port laparoscopic cholecystectomy and group B included the patients who would undergo LESS cholecystectomy. These patients had a BMI < 30, were ASA I or II, and had no previous upper-GI surgery. Blood was taken preoperatively and 6 and 24 h postoperatively. hsCRP (with automated analyzer) and α-defensins (using ELISA) were calculated for each sample. The same postoperative protocol was followed for both groups. Mann-Whitney U test was used to analyze the results. Pain was calculated with a visual analog scale (VAS) for shoulder and abdomen at 6 and 24 h. Hospital stay, nausea, and pain medication needed was noted. RESULTS: The α-defensins value was statistically significantly higher in the 24-h samples (P < 0.001) for LESS cholecystectomy. No statistically significant difference was shown for hsCRP, even though P = 0.05 for the 24-h samples with the values of LESS higher. No LESS was converted to a classical laparoscopic cholecystectomy, and none of the patients of either group needed conversion to open cholecystectomy. Pain was statistically significantly less for the LESS group at the 24-h interval (P < 0.0001). Less medication was needed for LESS patients after the 6th postoperative hour (P = 0.007). CONCLUSION: Higher inflammatory reaction in LESS cholecystectomy could be the result of greater tension on the tissues. More studies are needed to conclude if this has a significant clinical expression.

[Changes in liver function and immune function following laparoscopic and traditional open cholecystectomy: a comparative study].

OBJECTIVE: To compare the effect of laparoscopic cholecystectomy (LC) and open cholecystectomy (OC) on liver function and immune function. METHODS: Forty-two patients with normal liver function underwent LC (n=21) or OC (n=21) according to the patients' preference. One day before and 7 days after the operations, the liver functions (ALT, AST, TBIL, ALB) and immune functions (CD3, CD4, CD8, NK cell percentage, IgA, IgM, IgG and C3, C4) of the patients were measured. RESULTS: No statistical differences were found in the liver functions or immune functions between the two groups after the operation. CONCLUSION: LC and OC show no significant difference in the effects on the liver function and immune function.

CA19-9 antigen levels can distinguish between benign and malignant pancreaticobiliary disease.

BACKGROUND: CA19-9 is a carbohydrate tumor-associated antigen which is frequently upregulated in pancreatobiliary neoplasia. However, it may also be elevated in patients with jaundice in the absence of a tumor due to biliary obstruction, and in other non-hepato-pancreatico-biliary conditions. This study aimed to evaluate whether CA19-9 levels could accurately differentiate between benign and malignant pancreatobiliary disease. METHODS: All patients referred to a single surgeon for investigation of pancreaticobiliary disease in 2003 in whom a firm diagnosis had been established were included. For malignant disease, a histological diagnosis was required but for benign disease a firm radiological diagnosis was deemed adequate. The patients were divided into 4 categories: pancreatic adenocarcinoma (PCa); cholangiocarcinoma (CCa); chronic pancreatitis (CP) and biliary calculous disease (Calc). Bilirubin and alkaline phosphatase levels corresponding to the point of assessment of CA19-9 were also noted. RESULTS: Final diagnoses were made of pancreatic adenocarcinoma (PCa, n=73), cholangiocarcinoma (CCa, n=19), ampullary carcinoma (Amp, n=7), neuroendocrine carcinoma (Neu, n=4), duodenal carcinoma (Duo, n=3), chronic pancreatitis (CP, n=115), and biliary calculous disease (Calc, n=27). Median CA19-9 levels (U/ml) were: PCa, 653; CCa, 408; Duo, 403; Calc, 27; CP, 19; Neu, 10.5; Amp, 8 (reference range: 0-37). The CA19-9 levels were significantly greater for malignant than for benign disease, could differentiate PCa from CCa/Duo, and were significantly higher in unresectable than in resectable PCa. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for CA19-9 were 84.9%, 69.7%, 67.7% and 86.1%, respectively. A ROC analysis provided an area under the curve for CA19-9 of 0.871 (0.820-0.922), giving an optimal CA19-9 of 70.5 U/ml for differentiating benign from malignant pathology. Using this cut-off, the sensitivity was 82.1%, while specificity, PPV and NPV improved to 85.9%, 81.3% and 86.5%, respectively. When standard radiology was included (US/CT/MRCP) in the decision process, the results improved to 97.2%, 88.7%, 86.6%, and 97.7%. For benign disease, the CA19-9 correlated directly with the serum bilirubin, but for malignant disease, CA19-9 levels were elevated independent of the bilirubin level. CONCLUSIONS: CA19-9 is useful in the differentiation of pancreatobiliary disease and when using an optimized cut-off and combining with routine radiology, the diagnostic yield is improved significantly, thus stressing the importance of a multi-disciplinary approach to pancreatobiliary disease.

T-cell function is critical for murine cholesterol gallstone formation.

BACKGROUND & AIMS: The formation of cholesterol gallstones is a complex process involving contributions from genes and environmental factors. Although gallbladder inflammation is believed to be common during cholelithogenesis, the role of immunologic factors is unknown. METHODS: The role of adaptive immunity in cholesterol cholelithogenesis was analyzed utilizing immunocompetent Helicobacter spp.-infected and -uninfected BALB/c and congenic immunodeficient Rag2(-/-) (Rag) mice. Lymphocyte transfer studies were performed to determine which cellular subset was responsible for cholesterol gallstone formation. Also, gallbladder inflammation was quantified to determine the nature of the inflammatory response associated with cholelilithogenesis. RESULTS: When fed a lithogenic diet for 8 weeks, wild-type mice developed significantly more cholesterol gallstones (27%-80% prevalence) than Rag mice ( approximately 5%, P < .05). Helicobacter spp.-infected BALB/cJ mice displayed statistically significant increases in cholesterol gallstone prevalence compared with uninfected mice (81% vs. 39%; P < .05). Transfer of splenocytes or T lymphocytes to Rag2(-/-) mice increased stone prevalence markedly (26% and 40% respectively; P < .05), whereas transfer of B cells was not appreciably cholelithogenic (13%). The adaptive immune response increased the expression of gallbladder Muc genes and accumulation of mucin gel. In addition, T cells and cholesterol monohydrate crystals induced proinflammatory gene expression in the gallbladder, which likely contributes to gallbladder dysfunction. CONCLUSIONS: These studies indicate that T cells are critical in murine cholesterol cholelithogenesis. Furthermore, cholesterol monohydrate crystals induce expression of proinflammatory cytokines in a T-cell-dependent fashion. Acquired immunity and inflammation are likely to be crucial factors in cholesterol gallstone pathogenesis, rather then merely the result of cholelithogenesis.

[Changes of the different neuropeptide containing nerve elements in the inflamed human gall bladder]. / A küllözo neuropeptidtartalmú idegelemek számának változása humán gyulladt epehólyagban.

BACKGROUND, AIMS: The changes of different neuropeptide containing nerve elements might play a role in the pathogenesis of cholecystitis and the formation of gallstones, therefore the authors have investigated the density of the neuropeptide containing nerve fibres and immunocompetent cells in human gallbladder (control and cholecystitis). METHODS: The different neuropeptide containing nerve elements and immunocytes were detected by avidin-biotin-peroxidase (ABC) immunohistochemistry. RESULTS: In the control gallbladder the density of the different neuropeptide containing nerve fibres showed different pattern in all layers. In the inflamed gallbladder the number of the vasoactive intestinal polypeptide (VIP) positive nerve fibres increased significantly, very dense immunoreactive (IR) nerve fibres were located mainly in the tunica mucosa just below the epithelial lining. The number of the VIP IR nerve cell bodies was also increased. However, the number of the substance P (SP) IR nerve fibres was decreased significantly in the cholecystitis. The number of the neuropeptide Y (NPY) nerve fibres showed no changes, while their distribution was altered compared to the control. In the inflamed area the number of immunocompetent cells was strongly increased (being granulocytes, lymphocytes, plasma cells and mast cells) and some of them were also immunoreactive for SP, calcitonin gene-related peptide (CGRP) and VIP. Close contacts were detected between IR nerve fibres and the immunocytes in several cases. CONCLUSIONS: During inflammation the changes of the neuropeptide containing nerve fibres might alter the function (causing dilation) of the gall bladder, the activated immunocytes can also synthesize neuropeptides (SP, CGRP, VIP), so the released materials (cytokines, chemokines, histamine, as well as neuropeptides) might act in an autocrine and/or paracrine way influencing the function of the organ and of the immune system.

Gallbladder wall inflammatory cells in pediatric patients with biliary dyskinesia and cholelithiasis: a pilot study.

BACKGROUND/PURPOSE: Inflammation has been implicated in functional gastrointestinal disorders, including functional dyspepsia and irritable bowel syndrome. This study was undertaken to evaluate gallbladder wall inflammatory cells in children with abdominal pain related to gallstones and biliary dyskinesia to determine the candidate cell types that may be contributing to the pathophysiology of these entities. METHODS: Gallbladder specimens from 20 patients with cholelithiasis, 20 biliary patients with dyskinesia, and 12 autopsy controls were evaluated in a blinded fashion. Eosinophil, tryptase-positive, and CD3+ cell densities were determined for the lamina propria and muscularis mucosa layers and compared between groups. RESULTS: Patients with biliary dyskinesia and cholelithiasis had a 9- to 12-fold increase in mean and peak mast cell densities, respectively, in both layers as compared with controls. Peak (13.7 vs 8.4) and mean (9.2 vs 5.2) CD3+ cell densities were increased in the muscularis mucosae of cholelithiasis specimens as compared with biliary dyskinesia specimens. CONCLUSION: Gallbladder wall inflammatory cell densities, particularly mast cells, differ between children with cholelithiasis, children with biliary dyskinesia, and controls. Future studies are warranted to define the roles for specific inflammatory cell types.

[Characteristics of immune homeostasis in patients with cholelithiasis complicated by hepatitis and cholangitis].

The most distinctive and typical complications of cholelithiasis are cholangitis and hepatitis. The article presents an analysis of peculiarities of immune homeostasis in 41 patients with cholelithiasis complicated by hepatitis and cholangitis before and after surgical treatment and with the use of antihomotoxicologic and homeopathic medications.

Lethal hypercytokinemia following hepatic resection under pringle maneuver: a case report.

Clinical implications of acute reactant cytokines remain to be clarified in ischemia/reperfusion injury of humans. We report a lethal case of hypercytokinemia following continuous Pringle maneuver. A 36-year-old man with intrahepatic duct stones underwent left lobectomy under continuous hepatic inflow occlusion for 70 minutes. The postoperative course was stormy with rapid deterioration of liver functions, resulting in death due to multiorgan dysfunction on the 4th postoperative day. Analysis of cytokines demonstrated marked elevation of plasma acute inflammatory cytokines level (Interleukin-6 and -8) during surgery and immediate postoperative day. Our experience suggests that excessive production of inflammatory cytokines was detrimentally associated with multiorgan dysfunction including liver. The strategies against such hypercytokinemia should be considered when performing liver resection particularly under continuous Pringle maneuver.

Aspects of the humoral and cellular immune response in hepatitis associated with cholelithiasis.

AIM: To investigate the immunological reactions of the liver in various gall bladder disease. METHODS: IgG, IgA and IgM were measured in the blood and bile of patients with cholecystectomy. Immunofluorescence and electron microscopy techniques were used. RESULTS: The tests showed high blood values of immunoglobulins and their presence in the bile. The immunofluorescence was positive for IgG in the hepatic sinusoids. The electron microscopy showed aspects of "cellular cooperation". CONCLUSIONS: On the basis of our results we can affirm that in cholelithiasis there exists an immune response to the persistent antigens, with the release of circulating immune complexes.

Experimental model of acute pancreatitis in Wistar rat: glucocorticoid treatment profile.

Severe acute pancreatitis may be triggered by an extrapancreatic insult at the peri-Vaterian duodenum such as that occurring in the short-term, 20 min closed duodenal loop model in Wistar rat, which mimics biliary acute pancreatitis or that following endoscopy. Glucocorticoids are immunological modulators whose therapeutic value is worth investigating. Wistar male rats were used under standardized conditions. Acute pancreatitis was induced by instillation of a 7% sodium tauraocholate solution with 5 drops of methylene blue to monitor absence of duodenal bilio pancreatic reflux into the peri-Vaterian duodenum for 20 min. Detection of biliopancreatic reflux with methylene blue was an exclusion criterion. Different doses and times of administration of subcutaneous hydrocortisone were evaluated. Biochemical assays were carried out in blood samples and pancreatic and lung tissue, while histpathological studies were done in the pancreas, lung liver, duodenum, spleen, kidneys, suprarenal glands, and stomach. Animals subjected to the experimental model developed severe acute pancreatitis. According to the dose and time of administration, hydrocortisone therapy was effective and beneficial at a dose of 4 mg/kg give 30 min before inducing acute pancreatitis. It was ineffective when doses were <4 mg/kg and given before sodium taurocholate harmful when the dose was >4 mg/kg and given either before or after. Thus, the proposed model is valid and useful to study the initiation mechanism of acute pancreatitis caused extrapancreatically while its amelioration by glucocorticoid is related the dose and time factor to achieve therapeutical results.

Cyclooxygenase-2 mediates mucin secretion from epithelial cells of lipopolysaccharide-treated canine gallbladder.

Biliary mucin was regarded as a major contributing factor in formation of pigment stones as well as cholesterol ones. The aim of this study was to elucidate the mechanism of biliary mucin secretion in canine gallbladder epithelial (CGBE) cells treated by lipopolysaccharides (LPS) with special reference to cyclooxygenase (COX) -2. Confluent CGBE cells were incubated with following compounds for 8, 12, and 24 hr: (1) serum-free medium, (2) serum-free medium containing LPS (100 microm/ml), (3) serum-free medium containing LPS (100 microm/ml) with NS-398 (10 microM), and (4) serum-free medium containing LPS (100 microm/ml) with indomethacin (10 microM). Mucin assay and western blots for COX-1 and COX-2 were performed. Production of PGE2, and cAMP was also measured. Mucin secretion increased with time. At 12 hr, mucin secretion increased to 200% of control (from 100 +/- 5 to 200 +/- 45%, P < 0.05). LPS treatment significantly stimulated the COX-2 expression (P < 0.05). The productions of PGE2 and cAMP were increased from 299 +/- 68 to 524 +/- 163 pg/mg (P < 0.05) and from 0.2 +/- 0.1 to 0.92 +/- 0.4 pmol/ml (P < 0.05), respectively. NS-398, which completely inhibited COX-2 expression, significantly suppressed the level of PGE2 and cAMP as well as mucin secretion (P < 0.05). Indomethacin, which partially inhibited COX-2 expression, suppressed the production of PGE2, but not cAMP and mucin secretion. In conclusion, our results suggested that the PGE2 induced by COX-2 might play a role in mucin secretion from the gallbladder epithelium through the increment of cAMP.

Gram-negative bacteria killed by complement are associated with more severe biliary infections and produce more tumor necrosis factor-alpha in sera.

BACKGROUND: We previously showed that gallstones contain bacteria and that illness severity correlates with bacterial presence. This study examined virulence differences of gram-negative biliary bacteria. METHODS: Gallstones and bile were cultured, and sera obtained, from 210 patients. Infection severity was staged as: none-no clinical infection; moderate-fever, leukocytosis; or severe-bacteremia, cholangitis, hypotension, abscess, or organ failure. Gram-negative biliary bacteria were tested against patient (and control) serum for complement-mediated bacterial killing and induction of tumor necrosis factor-alpha (TNFalpha) production (using cultured monocytes) with and without sera. These results were correlated with infection severity. RESULTS: A total of 98 (47%) patients had biliary bacteria. Infection severity distribution was none, 29%; moderate, 35%; and severe, 36%. Gram-negative organisms killed by complement were associated with more severe infections as follows: 13%, none; 60%, moderate; and 88%, severe infections (P =.024 and P <.0001, respectively vs none, chi-square test). TNFalpha production in sera increased 182 pg/mL with complement resistant bacteria, but increased 546 pg/mL with bacteria killed by complement (P <.0001, killed vs not killed, Student's t test). E coli and Klebsiella were the most virulent bacterial species. They were cultured from blood, usually killed by complement, and had the largest increase in TNFalpha production in sera. CONCLUSIONS: Gram-negative biliary bacteria killed by complement (as opposed to complement-resistant) were associated with more serious biliary infections including bacteremia and induced more TNFalpha production in sera. This suggests a potential role for complement activation and cytokine production in biliary sepsis.