Thrombomodulin and antibeta2-glycoprotein I in stroke in children.

BACKGROUND: A quantitative systematic review recently discussed the role of thrombomodulin (Thm) and antibeta2-glycoprotein I (beta2-GPI) in cerebral strokes in adults. Little is known about the problem in children. The aim of the study was to see if there is a difference in the values of Thm and beta2-GPI in children with ischemic stroke. MATERIAL/METHODS: Seventy patients were included, comprising 40 children who had had ischemic stroke of unknown etiology hospitalized from January 1995 to December 2005 at the Department of Developmental Neurology, Chair of Neurology Medical University of Gdansk, and 30 healthy volunteers (no autoimmunologic disease or headache in interview). The concentrations of thrombomodulin (Thm) and antibeta2-glycoprotein I (beta2-GPI) in A, M, and G immunoglobulins were determined according to an immunoenzyme method (ELISA). RESULTS: None of the investigated subjects had elevated levels of beta2-GPI. The patients with stroke had significantly higher Thm values than the healthy group. CONCLUSIONS: This finding of elevated levels of thrombomodulin in cases of pediatric cerebral stroke could help in measuring the extent or duration of parenchymal brain injury, or even perhaps response to future therapeutic maneuvers. All these implications may aid not only in the diagnosis and management of acute ischemic stroke, but encourage prophylactic action to prevent probable stroke relapse.

Helicobacter pylori increases expression of proapoptotic markers Fas and FasL on CD4 lymphocytes in children.

The pathomechanism of Helicobacter pylori action upon gastric mucosa and its role in the pathogenesis of gastritis have not been fully elucidated. The aim of this study was to evaluate the most prevalent lymphocyte subpopulations of the gastric mucosa in gastritis in children, as well as to evaluate the expression of Fas and Fas ligand receptors (FasL), periapoptotic markers of gastric mucosa lymphocytes before and after H. pylori eradication. Forty nine patients aged 6 to 17 years, investigated due to chronic abdominal pain, were studied. The obtained tissue samples were analysed by immunohistochemistry. Different lymphocyte subsets were quantified on the basis of surface antigen expression (CD3, CD4, CD8, CD20), secreted cytokines (IL-4, IL-6, IFNgamma) and Fas and FasL proteins in the gastric mucosa. B and T helper lymphocytes were found to play a major role in the inflammatory infiltration in the gastric mucosa in children during H. pylori infection. Their expression was found to decrease after eradication. The enhanced expression of Fas receptor on lymphocytes before treatment and a decrease of this expression after eradication of H. pylori were shown. It was demonstrated that there is a correlation between CD4 and Fas receptor expression that may induce apoptosis of the helper lymphocytes in infected children.

[Periapoptotic markers in children with Helicobacter pylori infection]. / Markery okoloapoptotyczne w zakazeniu Helicobacter pylori u dzieci.

UNLABELLED: Helicobacter pylori (Hp) infection influences cell metabolism and apoptosis in the epithelium and lymphocytes of gastric mucosa. It may cause difficulties in the elimination of bacteria and lead to chronic gastritis. THE AIM OF THE STUDY was to find if there is a relationship between periapoptotic markers such as Fas, FasL and Bcl-2 in gastric mucosa in children with chronic gastritis and with Hp infection. MATERIAL AND METHODS: Forty-nine children with chronic abdominal pain were included in the study. They, were divided into three groups: group I without Hp (22) group II with (27) Hp infection. Eleven children from the second group who had follow-up endoscopy after eradication therapy formed group III. Hp infection was confirmed by 4 different methods. The triple- drug treatment was applied. Tissue samples from the gastric mucosa were obtained, during upper gastrointestinal endoscopy, for microscopic evaluation (according to the Sydney classification) and immunohistochemistry. In the analysed groups the percentage of patients with periapoptotic markers (Fas, FasL, Bcl-2) was established. The Fas antigen was estimated by immunofluorescent and immunoenzymatic methods. The FasL I Bcl-2 receptors were evaluated by the immunoenzymatic method. RESULTS: The expression of FasL and Bcl-2 receptors was found in all children without Hp infection. The expression of Fas antigen was less frequent in this group. The expression of Fas receptor was statistically significantly more frequent (p<0.05) in children with Hp infection. The expression of FasL and Bcl-2 in children with Hp infection was similar to group I (without Hp infection). In group of children after eradication treatment the expression of Fas and Bcl-2 (p<0.05) markers were significantly less frequent. CONCLUSIONS: Helicobacter pylori activates apoptosis by two pathways. It appears that the Fas/FasL pathway is the main one. Only in the case of Fas receptor there is a link between its significantly more frequent expression with Hp infection and its reduction after eradication treatment.

Expression patterns of Ki-67 and telomerase activity in middle ear cholesteatoma.

BACKGROUND: Keratinocytes in cholesteatoma demonstrate uncoordinated hyperproliferation, migration, and invasion properties. There is a controversy regarding the impact of Ki-67 and telomerase activities on cellular proliferation in cholesteatoma. We studied expression of Ki-67 protein and telomerase activity in cholesteatoma and its relationship with clinical findings. METHODS: The expression level of Ki-67 protein was examined by immunohistochemical analysis of 51 cholesteatomas and 6 skin tissues obtained from patients during ear surgery. Telomerase activity was determined in 23 samples of cholesteatomas and 6 skin samples by polymerase chain reaction-based telomeric-repeat amplification protocol assay. RESULTS: The presence of Ki-67 protein was observed in 21 (41.2%) of 51 samples of acquired cholesteatoma. The average Ki-67 labeling index in the cholesteatoma group was 28.9 +/- 9.2 and was higher than that in the skin group (18.2 +/- 6.1). Telomerase activity was detected in 2 (8.7%) of 23 samples of cholesteatoma (21 of them were Ki-67 staining positive and 2, negative) and in 3 (50%) of 6 of control skin samples (p < 0.05). CONCLUSION: This study showed increased expression of Ki-67 in cholesteatoma, whereas there was no significant difference in rate of Ki-67 positive staining between skin and cholesteatoma (p = 0.066). Telomerase activation is a rare event in cholesteatoma. We assume that the absence of telomerase may lead to generation dysfunctional telomeres what in turn may impair the proliferative capacity of cholesteatoma.

[Evaluation of selected immunocompetent cells expression in gastric mucosa in children with Helicobacter pylori infection before and after eradication treatment--a preliminary study]. / Ocena ekspresji wybranych komórek immunokompetentnych w blonie sluzowej zoladka u dzieci zakazonych Helicobacter pylori przed i po eradykacji zakazenia--doniesienie wstepne.

INTRODUCTION: Helicobacter pylori infection activates local immunological response and causes mononuclear cells infiltration in the gastric mucosa. On this account the studies on lymphocyte subpopulations in the gastric mucosa in children during Helicobacter pylori infections are inconsistent. It has been shown that the morphological status of gastric mucosa in children with Helicobacter pylori infection is different than in adult patients. THE AIM OF THE STUDY was the evaluation of chosen immunocompetent cells expression in gastric mucosa in children before and after eradication treatment. MATERIAL AND METHODS: Forty-nine children with chronic abdominal pain was enrolled in the study. They were divided into the following groups: 22 children without infection (negative urease test and absence of Helicobacter pylori antigen assessed by immunoenzymatic and immunofluorescent methods) and 27 with Helicobacter pylori infection. Part of the children (11) from the second group had a follow-up endoscopy after eradication therapy. The tissue samples from the gastric antrum and fundus were obtained for morphological and immunohistochemistry assays by direct immunofluorescent and immunoenzymatic methods. RESULTS: There were negative Helicobacter pylori tests in group I. In the group of infected children superficial colonisation of pathogen dominated In analysed groups percentage of patients with superficial antigens and cytokines (CD3, CD4, CD8, CD20, IL-4, IL-6, INF-gamma) characteristic for each lymphocytes subpopulations were established. In infammatory infiltrations T lymphocytes CD4 and B lymphocytes CD20 dominated localised mainly in the lamina propria of the gastric mucosa. Expression of above lymphocytes subpopulations diminished after eradication treatment. After treatment the total eradication of Helicobacter pylori was observed in 5 children and in 6 patients the pathogen persisted. CONCLUSIONS: The dominant role in local response during Helicobacter pylori infection in children is played by T CD4 and B CD20 lymphocytes localised mainly in lamina propria of gastric mucosa. Degree of T cells CD4 and CD20 expression decreases after eradication treatment.

[Assessment of selected haemostatic parameters in obese patients]. / Ocena wybranych parametrów ukladu hemostazy u osób otylych.

BACKGROUND: Epidemiologic data show significantly higher frequency of thromboembolic incidents in obesity than in normal weight persons. Disorders of haemostasis seem to play a crucial role in their development. In the literature there are only few papers assessing coagulation and fibrynolitic parameters in obese subjects. AIM OF THE STUDY: Assessment of protein C (PC), antithrombin (AT) and alpha2 antiplasmin (alpha2AP) activity and thrombomodulin (TM) concentration in the blood plasma of obese person (BMI > 30 kg/m(2)). MATERIALS AND METHODS: The study involved 32 patients (22 women and 10 men, mean age 39.7 +/- 15.3 years) and 20 healthy volunteers matched correctly according to sex and age who constituted a control group. In the examined subjects activity of PC, AT, alpha2 AP were assessed by means of the colorimetric methods and TM concentration in the blood plasma using ELISA method. RESULTS: No statistically significant differences in activities of PC, AT, alpha2 AP and TM concentrations between the patients and the control group were found. However the tendency to higher activities of PC and concentrations of TM were noticed in the obese patients. Assessing tested parameters according to sex, statistically significant differences were found in AT activity between the male patients and healthy men. Statistically lower values, but still in the normal range, were found in the obese men. Comparing the groups of women, significantly lower concentrations of TM were found in the obese ones. CONCLUSIONS: It seems that changed values of PC, AT, alpha2 AP activities are not responsible for increased risk of thromboembolic events in obese persons. Increased TM concentration in obese, may indirectly indicate endothelium damage.

[Antiphospholipid antibodies in children with migraine]. / Przeciwciala antyfosfolipidowe w migrenie u dzieci.

BACKGROUND AND PURPOSE: There are only a few investigations in the literature, that address the occurrence and the role of anticardiolipin antibodies (aCL) in children with migraine. The results of those studies are often contradictory. The aim of the study was to determine if the values of aCL in children with migraine differ from the control group. We tried to assess whether the type of migraine (with aura or without aura) had the influence on those values. MATERIAL AND METHODS: Sixty patients (mean age: 10.9+/-3.3 years), including 30 children with migraine hospitalized from January 2000 to December 2003 in the Department of Developmental Neurology Medical University of Gdansk and 30 healthy children, were studied. The values of aCL in class IgA, IgM and IgG were assessed by the immunoenzymatic method (ELISA test). RESULTS: The values aCL in IgA and IgG class were significantly different between the migraineurs and control group. The mean value of aCL in patients with migraine was 8.7+/-1.27 U/ml, while in the control group--3.81+/-1.74 U/ml. The positive values of aCL in class IgG were found in 11 (37%) children with migraine, and positive values of aCL in class IgM were noted in 6 (20%) cases in the same group. The type of migraine had no influence on the values of aCL. CONCLUSIONS: Children with migraine present with the higher values of aCL than the control group. The mean values of aCL were within the normal range, therefore their role in pathogenesis of migraine remains unclear. The further observation is needed to assess the reliable role of higher values of aCL in pathophysiology of vascular disorders.

Cell cycle inhibitory protein p27 in human middle ear cholesteatoma.

AIM: To evaluate the immunohistochemical and molecular presentation of protein p27 in cholesteatoma. METHODS: 42 cholesteatoma samples and 6 external ear canal skin (EECS) specimens were investigated and analyzed taking into consideration congenital, acquired, recurrent cholesteatoma, and EECS. RESULTS: The expression of p27 was found in 16 (38.1%) out of 42 specimens of cholesteatoma and in 5 (83.3%) out of 6 specimens of EECS. There was a significant difference in p27-positive staining rate between EECS and cholesteatoma epithelium (p < 0.008). The presence of p27 was detected in 10 cases of acquired cholesteatoma, 2 cases of congenital and 3 cases of recurrent cholesteatoma. There was no significant difference between the presence of p27 in cholesteatoma and EECS (p = 0.01). CONCLUSION: The down-regulation of p27 is a key player in cell cycle control and plays an undefined role in the pathogenesis of all types of cholesteatoma.

Interleukin-8 in the blood serum of patients with alcoholic liver disease.

BACKGROUND: The influence of the immune system on the development of alcoholic liver disease has recently been the object of attention. However, the connection between alcohol consumption, altered immune response, and development of changes in the liver has not been fully explained. The aim of the present study was to evaluate serum IL-8 concentration in patients with chronic alcoholic liver disease. MATERIAL/METHODS: 85 patients with different types of ALD and 35 healthy subjects were enrolled in the study. Serum IL-8 concentration was evaluated with the ELISA immunoenzymatic method. IL-8 in liver tissue was measured by the indirect immunofluorescence method. RESULTS: There was a significant correlation between IL-8 concentration and AST, ALP, GGT, total bilirubin and albumin levels in blood serum. A significantly higher concentration of IL-8 was seen in all the groups of ALD patients. The highest values were found in patients with chronic alcoholic hepatitis, and the lowest in those with fatty liver. Significantly higher values were found in patients with ascites or encephalopathy in comparison to those without any features of portal hypertension and/or insufficiency of the liver cells. A high concentration of the tested cytokine is a disadvantageous prognostic factor in patients with ALD. CONCLUSIONS: IL-8 appears to be an important factor in liver pathology in patients with ALD, especially in the development of the inflammatory process.

[Prognostic factors of celiac disease occurrence in type 1 diabetes mellitus children]. / Czynniki prognostyczne choroby trzewnej u dzieci z cukrzyca typu 1.

INTRODUCTION: Discussion on the frequency of coexistent celiac disease and type 1 diabetes mellitus (DM1) as well as an attempt to standardize diagnostic methods of celiac disease detection among DM1 children have been performed. OBJECTIVES: To assess the incidence of celiac disease among DM1 children in the Pomeranian region of Poland followed by analysis of the putative prognostic factors for celiac disease development in this particular group of children. MATERIALS AND METHODS: 70 children aged 9.47+/-4.59 (group 1) de novo diagnosed with DM1 and 223 children aged 10.20+/-3.87 with long-standing diabetes mellitus type 1 (4.47+/-3.16 years from the diagnosis) were enrolled in the study. All the patients had C-peptide, HbA1c, CRP, TSH, fT4, fT3, urinary albumin secretion rate, IgA, level of antigliadin antibodies (AGA), anti-tissue transglutaminase (TGA) IgA and IgG antibodies (ELISA), anti-endomysium (EmA) IgA and IgG antibodies (immunofluorescence) and anti-tyreoglobulin antibodies (TG), anti-thyroid peroxidase (TPO) antibodies (ELISA) evaluated. All the patients had jejunal biopsy and thyroid ultrasound examination. RESULTS: 5.7% of group 1 patients were diagnosed with celiac disease based on the positive jejunal biopsy in comparison with 9.4% in the group 2. TGA antibodies were present in 9.52% of group 2, AGA in 7.62%, EmA in 6.19%. 10% of group 1 children had autoimmune thyroiditis versus 24.2% of group 2 children. The group of children with coincident long-lasting DM1 and celiac disease (group A) was characterized by significantly earlier age at diagnosis (p=0.003), higher HbA(1)c (p=<0.001), CRP (p<0.001) and elevated urine albumin secretion in relation to children without celiac disease and autoimmune thyroiditis (group B). Serologic test detecting TGA antibodies was found to be the most sensitive (95.2%) for the detection of celiac disease among DM1 children, while the lowest sensitivity was obtained in the case of the EmA antibody test (61.9%). CONCLUSIONS: The celiac disease morbidity confirmed by jejunal biopsy is high among DM1 children (9.4%). The assessment of the serum TGA appears to be the most sensitive screening marker for the celiac disease detection in DM1 children.

[Portal vein thrombosis in patients with thrombophilia--own observations]. / Zakrzepica ukladu zyly wrotnej u chorych z trombofilia-- obserwacje wlasne.

Portal vein thrombosis is one of the main prehepatic causes of portal hypertension. The most frequent causes of thrombosis in this localization, apart from hepatic cirrhosis, are the following: acute inflammatory diseases and abdominal cancers, traumas, proliferative diseases of the hematopoietic system. In recent years attention was given to disorders in hemostasis, such as thrombophilia, in the course of which thrombosis development is particularly common. The authors present 10 patients after an incident of portal vein thrombosis, in which primary hepatic pathology was excluded and tests directed at thrombophilia were performed. In seven patients abnormalities in the examined parameters were found, and what is more, in two cases they had a complex character and involved more than one parameter. In five patients hyperhomocysteinemia was found. Among them, in two patients there was also a decreased protein S activity and in one of them there was also APC-resistance. In the next two patients there were abnormalities in one of the examined parameters - APC-resistance. Hyperhomocysteinemia was found in all patients with idiopathic thrombosis, and in one of them there were concurrent changes in protein S activity and APC-resistance. In patients with the history of portal vein thrombosis diagnostics of thrombophilia should be performed.

Detection of Helicobacter species in liver and stomach tissues of patients with chronic liver diseases using polymerase chain reaction-denaturing gradient gel electrophoresis and immunohistochemistry.

OBJECTIVE: Helicobacter DNA has been detected in the hepatobiliary tree of patients with chronic liver diseases (CLD). The presence of H. pylori in the stomach compared with in the liver of the same patients with CLD has not been studied, therefore to the aim of this study was to investigate the presence of Helicobacter DNA and antigens in the liver and stomach of Polish patients with chronic liver diseases using molecular and immunological methods. MATERIAL AND METHODS: Gastric mucosa and liver tissue samples and sera were collected from 97 Polish patients with CLD. Anti-H. pylori antibodies were detected by enzyme immunoassay (EIA), and H. pylori-like antigens detected by immunohistochemistry. Helicobacter DNA was detected in stomach and liver samples using a semi-nested Helicobacter genus-specific polymerase chain reaction (PCR) assay, and Helicobacter species identified by denaturing gradient gel electrophoresis (DGGE) and sequencing analysis of amplified PCR products. RESULTS: H. pylori was identified by DGGE and sequence analysis in 60/62 (97%) and 25/25 (100%) of the gastric and liver Helicobacter genus-positive samples, respectively, whereas DNA of H. heilmannii was detected in 2/62 (3%) of the Helicobacter genus-positive gastric samples. H. pylori cagA gene was detected in 23/62 (36%) and 3/25 (12%) gastric and liver tissue samples, respectively. H. pylori-like antigens were detected in 61/97 (63%) gastric mucosa and in 40/97 (41%) liver tissue samples. CONCLUSIONS: H. pylori-like organisms appeared to dominate the gastric mucosa and liver tissue of Polish patients with CLD. The prevalence of the cagA gene was higher in stomach compared with liver samples, which suggests a possible role of cagA negative H. pylori-like organisms in CLD. On the other hand, no significant correlation was found between the presence of H. pylori-like DNA and antigens in the liver and liver function tests.

Cytokines and clinical manifestations of malaria in adults with severe and uncomplicated disease.

Pro- and anti-inflammatory cytokines are supposed to be involved in malaria pathogenesis. Their relationship with clinical manifestations of the disease, however, is rarely studied in adults from non-endemic countries with imported disease, particularly with severe malaria. In this study we compared serum levels of gamma interferon (IFNgamma) and interleukins: IL-12, IL-18, IL-10 in healthy adults and patients with severe or uncomplicated imported malaria, with predominance of Plasmodium falciparum and Plasmodium vivax infections within studied group. Severe malaria was shown to be associated with elevated serum levels of IFNgamma and IL-18 as well as with relative deficiency of IL-12 mediated response in comparison to uncomplicated malaria cases, while IL-10 was found to be higher in all malaria patients compared to the controls. Overall, the results of our study are consistent with the observations from the regions with holoendemic malaria transmission, suggesting a pivotal role of impaired IL-12 expression in severe malaria. On the contrary, patients with severe malaria included into our study presented with the pattern of excessive production of inflammatory IFNgamma and IL-18, what seems to be an unusual finding compared to the results of the studies on African children and may be the feature of severe malaria in non-immune adults.

[Microstructure of tympanic membrane in chronic otitis media: immunohistochemical evaluation]. / Mikrostruktura blony bebenkowej w przewleklym zapaleniu ucha srodkowego w badaniach immunohistochemicznych.

Chronic otitis media (COM) is an inflammatory process involving the middle ear mucosa and the tympanic membrane. The healing and epidermization is mostly impaired by immunological response of the host. Investigating the activity and the function of immunological response elements one can learn the immunological mechanisms taking place in chronic otitis media. The ultrastructural investigations of the tympanic membrane were done on its fragments obtained from 19 patients with COM during middle ear surgery, performed at ENT Department of Medical University of Gdansk in the years 1997-1999. Immunohistochemical investigations were performed using monoclonal antibodies against tenascin, S-100 protein, Ki 67, CD 31, F VIII, HLA-DR, TGFbeta1 and EGFR. The control group was 11 healthy tympanic membranes from cadavers. The presence of tenascin was proven in all COM tympanic membranes and in 45.5% of those from control group. S-100 protein was present in 88.9% of the patients with COM and absent in control group. Ki 67 was observed in 44.4% of the patients with COM and in 27.3% of the healthy tympanic membranes. Angiogenesis factors (CD 31 and FVIII) were present in 77.8% of the investigated COM tympanic membranes, in control group in 45.5%. HLA-DR expression was observed in 90% COM patients, in control group in 72.7%. Growth factor TGFbeta1 was present in the all cases in mucous and fibrous layer and in 54.5% of healthy tympanic membranes. EGF receptor was present in 60% of COM patients, mainly in epithelial layer of tympanic membrane and in 54.5% of those from control group. The presented investigations confirm the immunological activity of tympanic membrane in chronic otitis media.

[Immunomorphological evaluation of cholesteatoma]. / Badania nad immunomorfologia perlakowego zapalenia ucha srodkowego.

Molecular and cellular mechanisms in chronic otitis media (COM) with cholesteatoma have not been clearly enough known so far. Investigations on cholesteatoma are focused on its immunological and morphological status. The authors presented the results of immunomorphological evaluation of 31 patients with COM with cholesteatoma, divided into three groups. In the first group there were 4 patients with congenital cholesteatoma, in the second group 19 patient with primary acquired cholesteatoma and in the third group 8 patient with secondary acquired cholesteatoma. Immunohistochemical investigations were performed using antibodies for identification the tenascin, S-100 protein, antigens Ki 67, CD 31, FVIII, HLA-DR, and growth factors TGFbeta1 and EGFR. The immunological activity was assessed for three types and matrix and perimatrix of cholesteatoma. High expression of tenascin was proven in the perimatrix of cholesteatoma and protein S-100 was highly expressed in the cholesteatoma matrix. Ki 67 antigen was seen rarely and mostly was present in basal cells of the matrix. The presence of endothelial cells was proven mainly in the connective and vascular tissue of perimatrix (CD 31, FVIII). The high expression of HLA-DR in matrix confirms the presence of Langerhans cells. The presence of growth factors TGFbeta1 and EGFR was observed in peribasal layer of the epithelium and keratinocytes of cholesteatoma matrix. Activity of immunological processes in COM with cholesteatoma confirms their important role in pathophysiology of chronic otitis media with cholesteatoma.

[Ocular immunology]. / Immunologia oka.

In this paper, some aspects of ocular immunological defence with particular consideration of regional immunity are reviewed. It is discussed the lacrimal drainage associated lymphoid tissue (LDALT) and unique factors determinating the eye as an immunologically "privileged" organ (blood-ocular barrier, sequestration of antigens, absence of lymphatic draining, ocular microenvironment modulating the immunity).

[Serum interleukin-6 concentrations in patients with alcoholic liver disease]. / Interleukina 6 w surowicy u chorych z alkoholow choroba watroby.

Serum interleukin-6 concentrations in patients with alcoholic liver disease Pathogenesis of alcoholic liver disease (ALD) is not well defined, but immune mediated hepatic injury is thought to be important. The main aim of this study was estimation of serum concentrations of IL-6 in patients with chronic alcoholic liver disease and determination of correlations between IL-6 serum concentration and occurrence of clinical manifestations, biochemical parameters and a stage of ALD. 85 patients with the diagnosis of chronic ALD and 35 healthy subjects (mached for age and sex) were enrolled into the study. Serum concentration of IL-6 was measured with ELISA. Serum IL-6 concentrations were markedly elevated in the all analyzed groups of patients with ALD when compared with healthy controls. When compared in groups, patients with alcoholic cirrhosis and chronic alcoholic hepatitis had the highest and patients with fatty liver had the lowest serum IL-6 concentrations. In addition, IL-6 concentrations were higher in patients with hepatic encephalopathy than in those without liver failure. Furthermore, we found statistically significant correlation between serum IL-6 and albumin concentrations. High IL-6 concentrations were associated with high mortality in patients with ALD. These findings suggest that IL-6 is an important immunological factor associated with alcoholic liver disease.

Autoimmune hepatitis in the material of Department and Regional Hospital of Infectious Diseases in Gdansk.

BACKGROUND: The aim of the study was the analysis of patients with autoimmune hepatitis (AIH), with respect to diagnostics, clinical course and treatment, based on the material from the wards of infectious diseases. MATERIAL/METHODS: The study was carried out in the group of 106 AIH patients--95 females aged 11-81 (mean age 46 years) and 11 males aged 8-73 (mean age 35 years). The diagnosis of AIH was based on international criteria, including biochemical test results, autoantibodies, and liver tissue morphology. Serological test excluded hepatitis of viral etiology. Diagnostic procedures included also blood cell count, biochemical parameters of liver function with protein fractions, immunoassays (immunoglobulins, autoantibodies), according to commonly used methods. Liver biopsies were performed in 93 patients. RESULTS: The clinical presentation mimicked acute viral hepatitis in 75% of cases, in the remaining 25% corresponded to chronic viral hepatitis. 26% had other coincident disorders of autoimmune etiology. In 84% the initial stage of the disease was characterized with moderately severe course, in 11%--severe, 7% of the patients died--half of them at the initial stage of the disease. The following morphological patterns of hepatic abnormalities were observed: hepatitis chronica agresiva, fibrosis periportalis, hepatitis chronica agresiva in cirrhosim vertens, cirrhosis hepatis activa, hepatitis chronica persistence, hepatitis granulomatosa. Over 40% of patients demonstrated relapses of the disease due to discontinuation of treatment after obtaining clinical and biochemical remission. 51 patients were treated with glucocorticosteroid monotherapy, the same number with glucocorticosteroids combined with azathioprine, 1 female patient underwent liver transplantation. In nearly 30% of patients, the diagnosis of AIH was established after a period of persistence of pathologic symptoms of over a year. CONCLUSIONS: Late diagnoses of AIH indicate insufficient knowledge of the disease among physicians. The methods of treatment used in AIH are not sufficiently effective. Discontinuation of treatment should be preceded in each case by overall assessment of the pathologic process, including biochemical parameters, autoantibody level and liver histopathology.

[Influence of erythropoietin on immunological system of patients with chronic renal failure]. / Wplyw erytropoetyny na uklad immunologiczny u chorych z przewlekla newydolnoscia nerek.

UNLABELLED: Recombinant human erythropoietin (epoetin) administration is a well established therapy of anaemia in maintenance hemodialysis (HD) patients. During the treatment, along with an increase in haemoglobin (Hb) level also an improvement of physical and sexual activity and cognitive functions was observed. Moreover, recent studies have shown an impact of epoetin on lipid-carbohydrate and protein metabolism, endocrinological functions and immune system [1-7]. It is still unclear whether all these changes are caused by direct epoetin activity or they are associated with the correction of anaemia and better oxygen supply and therefore an improvement of the conditions required for many metabolic function. The goal of the first study performed in our centre was to estimate the influence of epoetin alpha (Eprex) administered in the doses not affecting erythropoiesis (7-10 IU/kg/three times a week for 12 weeks) on serum levels of interleukin (IL) 2, 6 and tumor necrosis factor TNF-alpha [8, 9]. 10 HD patients (3 F, 7 M) aged from 33 to 62 years participated in that study. The level of IL-2, IL-6 and TNF-alpha was measured by means of bioassay using a highly sensitive cell line respectively CTLL, B9 and fibrosarcoma--WEHI 164 (clone 13). Cells viability was tested by colorimetric MTT assay. During the first period of observation stable Hb concentration and unchanged although significantly higher than in healthy people levels of TNF-alpha and IL-6 were noticed. Serum level of IL-2 increased significantly and in the 10 week it reached the values observed in healthy humans although after that period of time it dropped to the initial values. The aim of the following study was to estimate the influence of epoetin on IL-2, IL-10 and TNF-alpha production by whole blood cell culture [1, 2]. 10 HD patients (2 F, 8 M) aged from 35 to 53 years receiving standard doses of epoetin alpha (Eprex) for six months (in vivo experiments) and another 10 HD patients (3 F, 7 M) aged from 40 to 60 years not receiving epoetin participated in the study (in vitro experiments--cell culture were stimulated with different doses of (epoetin alpha--Eprex and epoetin beta--Recormon): 0.05; 0.1; 0.5; 1.0 IU/ml). IL 2 and TNF-alpha were measured using the bioassay mentioned above, IL 10 by ELISA immunoassay. The levels of IL10 increased in all epoetin treated patients and it was accompanied by transitory decrease of TNF-alpha. The levels of IL-2 increased in 7/10 patients under the study. Addition of epoetin in vitro to the whole blood culture of HD patients before implementation of epoetin confirmed that it is able to directly stimulate IL-2 production. The highest levels of stimulation of IL-2 secretion were observed for the physiological doses of epoetin (0.05 IU/ml). The aim of the more recent study was to examine changes in CD4+ and CD8+ T-cell subpopulations, the expression of the inhibitory molecule, CD152+ on T lymphocytes and the levels of IL-2, IL-6, IL-10, IL-12 and TNF-alpha in HD patients [10]. Additionally serum levels of C reactive protein (CRP), C3, C4 components of complement and immunoglobulin IgG, IgM and IgA were measured. 14 patients (8 F, 6 M) aged from 31 to 64 years receiving standard doses of epoetin alpha (Eprex) for twelve months (in vivo experiments) and another 4 HD patients (2 F, 2 M) aged from 43 to 57 years not receiving epoetin participated in the study (in vitro experiments--cell culture were stimulated with epoetin as in previous study). METHODS: IL-2, IL-6, and TNF-alpha were measured using bioassays described above, IL-12 and IL-10 by ELISA immunoassay. Expression of T-cell surface molecules was measured both in vivo by flow cytometry of lymphocytes sampled from peripheral blood and in vitro using whole blood cell culture stimulated with physiological as well as non-physiological doses of epoetin. The levels of C3, C4, IgG, IgM and IgA were estimated using nephelometric method. Compared with the findings before the start of epoetin therapy the CD4+/CD8+ ratio increased after 1 year of follow-up, whereas the percentage of CD152+ peripheral blood lymphocytes decreased. The increase of the CD4+/CD8+ ratio was dependent on a decrease of the percentage of CD8+ cells. The decrease of CD152+ population affected mainly CD8+152+ T cells. All these effects became apparent after 6 months of epoetin treatment. In vitro stimulation of whole blood cultures revealed that the addition of physiological concentration of epoetin decreased the percentage of CD8+152+ T cells. The pattern of the cytokines shifted towards Th1 phenotype (increase of IL-2 and IL-12) with a decreased level of proinflammatory cytokines (decrease of IL-6 and TNF-alpha). Treatment with epoetin did not alter plasma CRP, C3, C4 components of complement, immunoglobulin, as well as total count of lymphocytes. Summing up, administration of epoetin to maintenance HD patients not only treats the anaemia but also results in favourable changes in immune system. Epoetin is probably not only hemopoietic factor but also an immunomodulatory cytokine.