Long-term survival in HER2-positive metastatic breast cancer treated with first-line trastuzumab: results from the french real-life curie database.

BACKGROUND: Outcome of HER2-positive metastatic breast cancer (MBC) patients has improved since the use of trastuzumab. However, most HER2-positive MBC patients will progress within 1 year of trastuzumab-based therapy. Only limited data are available concerning long-term responders. METHODS: The primary objective of this study was to compare overall survival (OS) of HER2+ MBC patients with long-term response to first-line trastuzumab with overall survival of those with non-long-term response, based on two institutional databases: the French Epidemiological Strategy and Medical Economics program and the Breast Database. Long-term responders (LTR) were defined as patients with non-progressive disease for ≥ 2 years on first-line trastuzumab. Secondary objectives included progression-free survival (PFS), and predictive factors for LTR status. RESULTS: From 2004 to 2014, 422 HER2-positive MBC patients received first-line trastuzumab. With a median follow-up of 48 months, median OS and PFS were 63 months (CI95%, 50-71), and 18 months (CI95%, 15-21) respectively. In 111 patients (26.3%) classified as LTR, median OS was 110 months (CI95%, 95-not reached) versus 56 months in non-LTR patients (CI95%, 47-68). In multivariate logistic regressions, the following factors were independently associated with LTR status: number of metastatic sites (≤ 2 versus > 2, p = 0.01); endocrine therapy for metastatic disease (p = 0.001) and taxane-based first-line chemotherapy (p = 0.003). CONCLUSION: Several features are associated with long-term response to trastuzumab: few metastatic sites, taxane-based chemotherapy and maintenance endocrine therapy in HR+ patients. Further studies are needed to identify patients in whom trastuzumab can be stopped after several years of sustained response.

The Gene-Diet Associations in Postmenopausal Women with Newly Diagnosed Dyslipidemia.

OBJECTIVES: The aim of this study was to determine the relationship between polymorphisms of peroxisome proliferator activated receptor - PPAR gamma-2 (Pro12Ala, C1431T) and beta 3-adrenergic receptor - ADRB3 (Trp64Arg) and dietary habits in a group of postmenopausal women who were not under hypolipidemic treatment. DESIGN: Genetic, nutritional and anthropometric parameters were measured in 213 dyslipidemic (LDL ≥115 mg/dL) and 58 normolipidemic (LDL<115) postmenopausal women. The PCR-RFLP method were used to determine the distributions of selected alleles and genotype frequencies. Dietary intake of basic components and fatty acids was obtained from a 7-day weighed food record and the bio-impedance method was used to determine nutritional status. RESULTS: Nearly 79% of analyzed women were in the first-time-diagnosed dyslipidemic state. The dyslipidemic subjects were characterized with higher intake of energy, fat, and saturated fatty acids (SFA). The analysis of the same polymorphisms showed association at the P value <0.05 with nutrients (fat, SFA, and polyunsaturated fatty acid - PUFA and saccharose) and elevated LDL level. Higher PUFA intake in a group of women with the protective Ala12/X polymorphism did not increase the risk of dyslipidemia even though they were characterized by visceral distribution of fat. The Arg64/X polymorphism and higher intake of energy, fat, and arachidic acid intake (C20:0) were associated with dyslipidemic state. CONCLUSION: Both nutritional and genetic factors are related to lipid profile. The identification of gene-diet associations is likely to provide useful information about the etiology of postmenopausal dyslipidemia and help in effective treatment.

Analysis of immunohistochemical expression of proinflammatory cytokines (IL-1α, IL-6, and TNF-α) in gallbladder mucosa: comparative study in acute and chronic calculous cholecystitis.

BACKGROUND: Several studies have shown increased serum levels of proinflammatory cytokines (IL-1α, IL-6, and TNF-α) in patients with cholelithiasis. The local expression of the proteins involved in pathogenesis of the disease is poorly recognised. MATERIALS AND METHODS: The authors examined immunohistochemically (IHC) the expression status of IL-1α, IL-6, and TNF-α in gallbladder mucosa of the patients with cholelithiasis as related to acute (ACC) and chronic (CCC) types of cholecystitis. Proinflammatory cytokines were quantitatively evaluated in gallbladder mucosa (epithelium and lamina propria) in ACC (n = 16) and CCC (n = 55) groups using modern spatial visualisation technique. RESULTS: Quantitative analysis of IHC signals showed no significant differences in IL-1α and IL-6, and immunoexpression in patients with ACC and CCC. A significantly greater IHC expression of TNF-α was detected in CCC as compared with ACC group. In either of the patient groups immunoexpression of IL-1α and of TNF-α was significantly higher than that of IL-6. Immunoexpression of TNF-α was significantly higher than that of IL-1α only in CCC group. A positive correlation was disclosed between IHC expression of IL-1α and body mass index in CCC group. IHC expression of TNF-α correlated positively with expression of CD68 molecule (histiocytic marker), number of leukocytes in blood and higher grading of gallbladder wall in ACC group. CONCLUSIONS: A more pronounced IHC expression of TNF-α and IL-1α than IL-6 in both types of cholecystitis may suggest the role of these cytokines in pathogenesis of cholelithiasis. IHC expression of TNF- α shows better correlation with clinical/laboratory data in acute cholecystitis, and its quantitative prevalence over the remaining cytokines points to the role of the TNF-α in maintenance of inflammation in the course of cholelithiasis.

Discordant expression of desmoglein 2 and 3 at the mRNA and protein levels in nodular and superficial basal cell carcinoma revealed by immunohistochemistry and fluorescent in situ hybridization.

BACKGROUND: Basal cell carcinoma (BCC) is the most common human cancer. It is thought that skewed expression of desmogleins (Dsgs) in BCC may promote tumourigenesis. AIM: To comparatively examine expression of Dsg2/Dsg3, using fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) in BCC subtypes. METHODS: In total, 84 frozen sections from patients with various clinical or histological subtypes of BCC were analyzed. Expressions of Dsg2/Dsg3 protein and Dsg2/Dsg3 mRNA were evaluated using IHC and FISH, respectively, in BCC nests and BCC-free epidermis, and then quantitatively measured. RESULTS: There was loss of correlation between Dsg2 and Dsg3 (IHC) in nodular and superficial BCC (nBCC, sBCC), and significant correlation between Dsg2 and Dsg3 (FISH) in BCC, but not nBCC and sBCC. CONCLUSIONS: Because more prominent aberrations of Dsg2/Dsg3 expression were seen at the protein than at the mRNA level in BCC, these comparative observations indicate greater importance of events at the proteome level than those at the genome level in tumour functional compartments. Different Dsg2/Dsg3 expression in sBCC and nBCC might corroborate the possibility that sBCC and nBCC are separate conditions. These results may contribute to better understanding of the biological behaviour of BCC.

Expression of phenotypic markers of mast cells, macrophages and dendritic cells in gallbladder mucosa with calculous cholecystitis.

The study aimed at quantitative analysis of expression involving markers of mast cells (tryptase), monocytes/macrophages (CD68 molecule) and dendritic cells (S100 protein) in gallbladder mucosa with acute and chronic calculous cholecystitis. Routinely prepared tissue material from the patients with acute (ACC) (n = 16) and chronic calculous cholecystitis (CCC) (n = 55) was evaluated. Three cellular markers were localized by immunocytochemistry. Their expression was quantified using spatial visualization technique. The expression of tryptase was similar in acute and chronic cholecystitis. CD68 expression in ACC was significantly higher than in the CCC group. Expression of S100 protein was significantly higher in CCC as compared to the ACC group. No significant correlations were disclosed between expression of studied markers and grading in the gallbladder wall. A weak negative correlation was noted between expression of CD68 and number of gallstones in the CCC group. The spatial visualization technique allowed for a credible quantitative evaluation of expression involving markers of mast cells (MCs), monocytes/macrophages (Mo/Ma) and dendritic cells (DCs) in gallbladder mucosa with ACC and CCC. For the first time mucosal expression of S100 protein-positive DCs was evaluated in calculous cholecystitis. The results point to distinct functions of studied cell types in the non-specific immune response in calculous cholecystitis.

Microencapsulated sodium butyrate reduces the frequency of abdominal pain in patients with irritable bowel syndrome.

AIM: Abdominal pain, defaecation disorder and change of bowel habit are the commonest symptoms of irritable bowel syndrome (IBS). The effect of microencapsulated sodium butyrate (MSB) was assessed on the severity of symptoms in patients with IBS. METHOD: Sixty-six patients treated with one of the standard pharmacological therapies for at least 3 months were included in the study. They were randomized to receive MSB as a supplemental treatment to standard therapy or to receiving a placebo. Previous pharmacological therapy was continued throughout the study in both arms. Clinical evaluation was performed at baseline, 4 and 12 weeks. Each assessment was documented by a validated visual analogue score questionnaire measuring the severity of selected clinical symptoms, a closed-end questionnaire measuring the frequency of selected clinical symptoms and a single closed-end question measuring the subjective improvement of symptoms. RESULTS: After 4 weeks there was a significant decrease of pain during defaecation in the MSB group which extended to improvement of urgency and bowel habit at 12 weeks. Reduction of abdominal pain, flatulence and disordered defaecation was not statistically significant. CONCLUSIONS: MSB as a supplemental therapy can reduce the frequency of selected clinical symptoms in patients with IBS, without significant influence on reducing symptom severity.

Pouchitis may increase the risk of dysplasia after restorative proctocolectomy in patients with ulcerative colitis.

AIM: Dysplasia of the pouch mucosa after restorative proctocolectomy is rare. The aim of this study was to establish whether there is a correlation between pouchitis and dysplasia. METHOD: A group of 276 patients treated for ulcerative colitis by restorative proctocolectomy between 1984 and 2009 was analysed. The presence or absence of pouchitis and dysplasia within the pouch was evaluated. RESULTS: Inflammation was diagnosed in 66 (23.9%) patients, low-grade dysplasia in five (1.8%), high-grade dysplasia in three (1.1%), and cancer in one patient (0.4%). The prevalence of low-grade dysplasia was significantly higher in patients with inflammation than in those without (P < 0.04). High-grade dysplasia was significantly more frequent in pouchitis than in non-inflamed pouches (P < 0.01). Logistic regression analysis suggested that the occurrence of mucosal inflammation increased the risk of low grade dysplasia. CONCLUSION: Patients with chronic pouchitis are at risk of dysplasia and require surveillance of the pouch.

The influence of elastic components of the venous wall on the biomechanical properties of different veins used for arterial reconstruction.

OBJECTIVE: To evaluate the relationship between the biomechanical properties and the structure of elastic components in different veins used for vascular reconstruction. DESIGN: In vitro experimental study. MATERIAL AND METHODS: Groups of 30 samples of incompetent saphenous veins (rSV), competent saphenous veins (cSV) and femoral veins (FVs) were compared following immunohistochemical staining for the presence of collagen types I, III and IV and elastin. The percentage area of transverse section of veins occupied by each type of collagen and elastin was measured using a computer-image-analysis system connected to a microscope. For all three groups of veins, the storage modulus, E', and the loss modulus, E'', were measured with a mechanical analyser, DMA-242, and changes in the function of temperature and frequency, and duration of exposure to the applied force were determined. RESULTS: The rSV showed the highest percentage share of collagen I and the lowest percentage share of collagen IV. These samples also showed the greatest expression of elastin and the highest elastin to collagen ratio. The rSV were also found to have the highest E' and E'', and during the long-term exposure achieved maximum stiffness in the least time as compared to cSV and FV. CONCLUSION: The histological structure directly influences the biomechanical properties of venous wall with rSV showing least compliance and cSV the greatest compliance.

Overexpression of CD39/nucleoside triphosphate diphosphohydrolase-1 decreases smooth muscle cell proliferation and prevents neointima formation after angioplasty.

BACKGROUND: Growing evidence implicates the involvement of extracellular nucleotides in the regulation of platelet, leukocyte, endothelial cell (EC) and vascular smooth muscle cell (VSMC) phenotype and function. Within the quiescent vasculature, extracellular nucleotides are rapidly hydrolyzed by CD39, the dominant endothelial nucleoside triphosphate diphosphohydrolase (NTPDase-1). However, vascular CD39/NTPDase-1 activity is lost in EC activated by oxidative stress or proinflammatory mediators, and upon denudation of the endothelium following balloon injury. The consequent increase in extracellular nucleotide concentrations triggers signaling events leading to prothrombotic responses and increased VSMC proliferation. OBJECTIVES: To investigate the effect of overexpressed CD39/NTPDase-1 in injured aorta. METHODS: Using adenoviral-mediated gene transfer we expressed CD39/NTPDase-1 in mechanically denudated rat aortas. We measured intima formation by morphometry and VSMC proliferation by the [(3)H]-thymidine incorporation assay. RESULTS: Targeted expression of CD39 in injured vessels increased NTPDase activity (from 2.91 +/- 0.31 to 22.07 +/- 6.7 nmols Pi mg(-1) protein, 4 days after exposure to the adenovirus) and prevented the formation of neointima. The thickness of the intimal layer in injured aortas exposed to Ad-CD39 was 26.2 +/- 3.9 microm vs. 51.8 +/- 6.1 microm and 64.4 +/- 22.2 microm (P < 0.001) in vessels treated with Ad-beta-gal and saline, respectively. Moreover, targeted expression of CD39/NTPDase-1 caused a 70% (P < 0.01) decrease in proliferation of VSMC isolated from transduced rat aortas as compared with VSMC derived from control vessels. CONCLUSIONS: The presented data suggest that increasing CD39/NTPDase-1 activity in VSMC could represent a novel therapeutic approach for the prevention of stenosis associated with angioplasty and other occlusive vascular diseases.

[Neurolymphomatosis. Subacute sensorimotor polyneuropathy as a first sign of non-Hodgkin's B cell lymphoma]. / Neurolymphomatose. Subakute sensomotorische Polyneuropathie als Erstmanifestation eines Non-Hodgkin-B-Zell-Lymphoms.

We report on a 65-year-old patient with subacute painful sensorimotor polyneuropathy with distal leg muscle palsy and initially presenting with bilateral leg edema. Electrophysiologic testing showed an axonal lesion pattern with acute denervation. Nerve biopsy demonstrated neurolymphomatosis as an initial manifestation of a non-Hodgkin's B cell lymphoma.

Immunohistochemical detection of galectin-1 in renal biopsy specimens of children and its possible role in proteinuric glomerulopathies.

AIMS: Galectin-1 is an endogenous lectin that specifically binds to beta-galactoside structures. It has been associated with developmental mechanisms ranging from differentiation to apoptosis and exerts immunoregulatory functions in autoimmune diseases. The aim was to determine the immunohistochemical expression of galectin-1 in renal biopsy specimens of children with primary idiopathic proteinuric glomerulopathies. METHODS AND RESULTS: We examined 18 children with minimal change disease (MCD), 30 with diffuse mesangial proliferation (DMP) and 11 with focal segmental glomerulosclerosis (FSGS). An indirect immunohistochemical protocol using a polyclonal antibody directed against galectin-1 was applied. Galectin-1 was detected in renal podocytes in DMP and FSGS cases, while control glomeruli and MCD were negative. Galectin-1 immunoreactivity was found within parietal epithelial cells in patients with FSGS. CONCLUSIONS: These results suggest a possible role for galectin-1 in the pathogenesis of primary glomerulopathies in children as a kind of podocyte-related self-protective activity and probably involvement of epithelial cells of Bowman's capsule in inflammatory processes. Immunohistochemistry using galectin-1 antibodies may further be helpful in histological distinction between MCD and DMP.

Vascular endothelial growth factor C--a potent risk factor in childhood acute lymphoblastic leukaemia: an immunocytochemical approach.

AIMS: To investigate the immunocytochemical expression of vascular endothelial growth factor C (VEGF-C) and its receptors (VEGFR-2 and VEGFR-3) in childhood acute lymphoblastic leukaemia (ALL) blasts and to determine the possible role of this complex in the pathogenesis and prognosis of ALL. METHODS AND RESULTS: Bone marrow samples were taken from 120 children diagnosed with ALL. An indirect immunocytochemical procedure was performed with the use of monoclonal mouse anti-human antibodies against VEGF-C, VEGFR-2 and VEGFR-3 (diluted 1 : 100). The immunocytochemical expression of VEGF-C was confirmed exclusively in the cytoplasm of ALL lymphoblasts (the mean percentage was 36.4 +/- 7.2). It was absent from the cytoplasm of normal haematopoietic cells in the control group. No VEGFR-2 or VEGFR-3 expression was detected in the children of either the study or control groups. The risk of induction failure or leukaemic relapse was found to be significant in all VEGF-C+ patients (P < 0.0001 and P < 0.02, respectively; Fisher's exact test). CONCLUSIONS: The absence of VEGF-C in blast cells predicts long-lasting remission in all leukaemic children. Our findings also suggest that leukaemic cell invasion, following VEGF-C-driven lymphangiogenesis, could be related to a mediating role of this peptide produced by blast cells themselves.

Antibodies to tissue-type plasminogen activator (t-PA) in patients with inflammatory bowel disease: high prevalence, interactions with functional domains of t-PA and possible implications in thrombosis.

BACKGROUND: Patients with inflammatory bowel disease (IBD) have an increased prevalence of thromboembolic events. The pathogenetic mechanisms of these events include reduced fibrinolysis, which may be caused by antibodies to tissue-type plasminogen activator (t-PA). OBJECTIVES: To evaluate anti-t-PA antibodies in patients with IBD, considering clinical, biochemical and functional characteristics. PATIENTS AND METHODS: We immunoenzymatically measured anti-t-PA antibodies in plasma from 97 consecutive IBD patients and 97 age- and sex-matched healthy controls. We also assessed the antibody interactions with different epitopes of t-PA, the antibody inhibition on t-PA activity and the correlations with clinical features and other serum antibodies. RESULTS: IBD patients had higher median anti-t-PA antibody levels (5.4 U mL(-1) vs. 4.0 U mL(-1); P < 0.0001): 18 patients were above the 95th percentile of the controls (OR 5.3; 95% CI 1.7-16.3; P < 0.003), and the six with a history of thrombosis tended to have high levels (6.9 U mL(-1)). Anti-t-PA antibody levels did not correlate with IBD type, activity, location or treatment, or with age, sex, acute-phase reactants or other antibodies. The anti-t-PA antibodies were frequently IgG1 and bound t-PA in fluid phase; they recognized the catalytic domain in 10 patients and the kringle-2 domain in six. The IgG fraction from the three patients with the highest anti-t-PA levels slightly reduced t-PA activity in vitro. CONCLUSIONS: The prevalence of anti-t-PA antibodies is high in IBD patients. By binding the catalytic or kringle-2 domains of t-PA, these antibodies could lead to hypofibrinolysis and contribute to the prothrombotic state of IBD.

Significance of alterations in PBMC immunophenotype of children with chronic viral hepatitis C-- the role of dendritic cells.

There are differences in the clinical course of chronic viral hepatitis C between adults and children, but it is generally accepted that the disease has cell-mediated immune background. The aim of this study was to evaluate PBMC subsets in children with chronic hepatitis C before treatment in order to find some predictive factors, useful for patients management. Several PBMC subsets, in particular lymphoid and dendritic cell (DC) ones, were tested by flow cytometry in HCV(+) paediatric patients (n = 46) and in control children matched in terms of age and sex (n = 20). Data were subjected to extensive statistics. It was found that cells with cytotoxic potential such as CD8(+)CD28(-) T cells, NK and NKT cells as well as lineage(-)HLA-DR(+) DC were increased in per cent values, while CD4(+) T cells and CD4:CD8 ratio were decreased in hepatitis C group. In HCV(+) patients, CD4(+) T cells were inversely correlated with alanine aminotransferase (ALT) levels and with viraemia. DC subset of myeloid origin (CD11c(+)) assessed both in per cent values and as mean fluorescence intensity (MFI) of HLA-DR expression was shown to be downregulated in hepatitis patients, in spite of increased numbers. To conclude, PBMC subsets, and in particular DC, are affected by HCV chronic infection in children, reflected by the correlation with clinical parameters, such as ALT and viraemia.

Ezrin--a useful factor in the prognosis of nephrotic syndrome in children: an immunohistochemical approach.

BACKGROUND: Minimal change disease (MCD) and diffuse mesangial proliferation (DMP) are the most common pathomorphological forms of nephrotic syndrome glomerulopathies in children. The clinical course of DMP can be characterised by either DMP-sensitivity (DMP-S) or DMP-resistance (DMP-R) to steroids, resulting in an unfavourable course of the glomerulopathy. Although the clinical processes of DMP-S and DMP-R are initially identical, resistance to steroids may be foreseen by the immunohistochemical expression of cytoskeleton-associated proteins in podocytes. AIMS: To estimate the immunohistochemical expression of ezrin in children with MCD, DMP and focal segmental glomerulosclerosis (FSGS) and to evaluate its usefulness in predicting resistance to steroids. MATERIALS AND METHODS: Renal biopsy specimens of patients with MCD (n = 15), DMP (n = 16) and FSGS (n = 6) were taken. The control tissue consisted of normal-appearing cortex taken from kidneys resected for localised neoplasms (n = 6). The indirect immunohistochemical protocol for the use of a monoclonal antibody directed against ezrin was used. RESULTS: The immunohistochemical expression of ezrin in cases progressively reduced from MCD to DMP-S to DMP-R to FSGS. Except for DMP-R and FSGS (p>0.05), the difference in ezrin expression in podocytes was significant. CONCLUSION: Ezrin can be a potent marker of podocyte injury (podocytopathy) and may help in the histological qualification of MCD, DMP and FSGS. The increased permeability of the filtration barrier in steroid-resistant and proteinuric glomerulopathies may be a consequence of subcellular changes in podocyte-associated proteins following decreased expression of ezrin.

Vascular endothelial growth factor (VEGF-C1)-dependent inflammatory response of podocytes in nephrotic syndrome glomerulopathies in children: an immunohistochemical approach.

AIMS: To analyse expression and distribution of vascular endothelial growth factor (VEGF-C1), podocalyxin and synaptopodin within renal tissue in nephrotic syndrome glomerulopathies in children. METHODS AND RESULTS: Renal biopsies performed at the time and in the manner recommended by the World Health Organization. The study group consisted of submicroscopic glomerulonephritis (n = 10), diffuse mesangial proliferation (n = 14) and focal segmental glomerulosclerosis (n = 5). The control tissue consisted of macroscopically normal appearing cortex taken from kidneys resected for localized neoplasms (n = 3). Material for immunohistochemistry was fixed in Bouin's solution and embedded in paraffin. Indirect immunohistochemistry using monoclonal anti-human antibodies directed against VEGF-C1, podocalyxin and synaptopodin was employed. The distribution of markers was quantified by computerized image analysis. In non-sclerosed glomeruli (within podocyte cytoplasm), VEGF-C1 was more expressed in podocytes of all groups (P < 0.0002), while the distribution of synaptopodin was less expressed in all groups (P < 0.0002). There was no statistical difference between all groups in the expression of podocalyxin. CONCLUSIONS: The increased permeability of the filtration barrier in steroid-resistant glomerulopathies may be a consequence of subcellular changes in podocytes resulting from decreased expression of synaptopodin. Moreover, impaired permeability of endothelium could be secondary to increased expression of podocyte-derived VEGF-C1.

Morphometrical analysis of immunohistochemical reaction of inflammatory infiltrate in chronic thyroiditis.

The aim of the study was to quantitatively evaluate B and T lymphocytes and macrophages, based on immunohistochemical investigations (CD43, CD20, CD8 and CD68) of chronic focal and Hashimoto thyroiditis. A new method of image analysis was applied, based on spatial visualization of the antigens reactivity. The obtained results indicated that the numbers of lymphocytes, in particular of cytotoxic T lymphocytes, and of macrophages increased with the progress of inflammatory process. Quantitative measurements of the markers made the results more objective and supported pathomorphological diagnosis.

Techniques of image analysis for quantitative immunohistochemistry.

The aim of this paper was to evaluate the usefulness of digital image analysis techniques to measure the amount and strength of immunohistochemical markers. The new method, based on the spatial visualization technique, was confronted with methods of colour sampling and grey scale thresholding. Examples of applications of the techniques for apoptosis and proliferation markers are also presented.

Quantitative evaluation of immuno- and histochemical reaction intensity by spatial visualization techniques.

The aim of this study was an application of spatial visualization techniques for quantitative measurements of immuno- and histochemical reactions. For a quantitative histochemical study, specimens, collected from patients with chronic gastritis, were stained with paS/AB, while for immunohistochemical evaluation, specimens were used, collected from patients with chronic parathyroiditis and were analyzed with Ki-67 proliferation marker and apoptosis bcl-2 protein. The new technique permitted to obtain quantitative objective results. Statistical cluster analysis of those results indicated small groups of cases for reevaluation and supported the final diagnosis.

Intracellular expression of pro-inflammatory cytokines (IL-1alpha, TNF-alpha, and IL-6) in chronic hepatitis C.

The study aimed at localizing TNF-alpha, IL-1alpha, IL-6 at light and electron microscope levels in patients with chronic hepatitis C, using the immunocytochemical techniques in biopsy material from patients with chronic hepatitis C and at comparing the expression of the cytokines with histopathological changes. Our studies demonstrated an augmented expression of all cytokines in liver biopsies in chronic hepatitis C, in comparison with respective values, obtained in control biopsy material. The highest expression of the cytokines was observed in hepatocytes. That was confirmed by electron microscopy, which demonstrated the cytokines mainly in altered ER cisterns and in the cytoplasm. In children, the expression of IL-1alpha was negatively correlated with staging, while in adult patients; the staging was positively correlated with the expression of TNF-alpha. The new element involves demonstration of cellular and subcellular expression of TNF-alpha, IL-1alpha and IL-6 in hepatocytes in in vivo infection.