Seronegative hepatitis C virus infection in Polish blood donors-Virological characteristics of index donations and follow-up observations.

Nucleic acid testing (NAT) was implemented in Poland in 1999 for screening of plasma for fractionation and for all blood donors in 2002. To analyze seronegative NAT-positive samples representing hepatitis C virus (HCV) window-period (WP) in the years 2000 to 2016 and to determine infection outcome. We analyzed results of 17 502 739 donations screened in minipools (6-48) or individually. Index samples underwent viral load (VL) quantification, genotyping and Ag, and anti-HCV re-testing using chemiluminescence (CMIA), electrochemiluminescence (ECLIA), and fourth-generation enzyme-linked immunosorbent assay (IV EIA) assays. HCV-seronegative infections were identified in 126 donations (7.2/mln donations; 95% confidential intervals, 6.0-8.6). Frequency of NAT yields was decreasing over time. Of the initial 126 seronegative index cases 106 were retested: 32.1% were reactive in IV EIA, 11.3% in ECLIA, and 1.9% in CMIA. The lowest VL correlated with absent anti-HCV and HCV Ag, while VL was highest when the antigen was detectable and then it decreased when anti-HCV appeared at a level detectable by sensitive third generation tests while retesting. The proportion of genotype 1 was 38.9% in samples positive only for HCV RNA and 71.4% in samples that were anti-HCV reactive in re-testing. In parallel, genotype 3 frequency was 50% in the former group and 21% in the latter. NAT is an effective measure to limit HCV transmission by transfusion and IV EIA seems to have higher clinical sensitivity than ECLIA. Samples representing likely successive phases of early HCV infection were characterized by different genotype distribution probably due to very early elimination of genotype 3.

RNA-HCV viral load in serum, peripheral blood mononuclear cells and liver in children with chronic hepatitis C.

The liver is the major site of hepatitis C virus (HCV) infection and replication. However, HCV may infect and replicate in extrahepatic sites as well. Several investigators have demonstrated that peripheral blood mononuclear cells (PBMCs) are the major extrahepatic milieu of infection and viral replication. The aim of the study was to investigate the correlation between RNA-HCV level in serum. PBMCs and liver in children with chronic viral hepatitis C (CHC). The impact of RNA-HCV level on the sustained virological response (SVR) after therapy was also determined. Study was carried out in the group of 10 children with CHC, age 8 to 17 years. Antiviral therapy was implemented in all patients with pegylated interferon alpha (Peg-lFNalpha) 2a or 2b and ribavirin during 48 weeks. The following tests were performed prior the therapy: basic laboratory parameters, histology of liver biopsy, RNA-HCV viral load in serum, PBMCs and in liver. The behavior of HCV-RNA viral load in serum, PBMCs and liver in children with CHC did not present strict mutual relations. However, the positive correlation between serum and PBMCs viral load (r = 0.47) and negative correlation between PBMCs and liver viral load (r = -0.47) was demonstrated. Although no statistically significant results were found, some trends of relationship in viral load between various body compartments were present. Given the aforementioned results, it is clear that more data are needed, mostly more numerous groups of patients, especially those whose influence of RNA-HCV viral load had a major impact on the antiviral treatment.

Is interleukin-8 an additional to histopathological changes diagnostic marker in HCV-infected patients with cryoglobulinemia?

PURPOSE: The aim of this study was to analyze histopathological changes in the liver and serum inflammatory cytokine level in hepatitis C virus (HCV)-infected patients with and without cryoglobulinemia. METHODS: The study group consisted of 34 patients with chronic hepatitis C, confirmed by serological and virological markers. Ten out of 34 patients had cryoglobulinemia. The control group consisted of 21 healthy persons. Liver biopsy specimens of HCV-infected patients were evaluated by light microscopy using the grade and the stage according to Batts and Ludwig classification. The quantitative measurements of IL-1ß, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor in sera were performed by flow cytometry. RESULTS: The mean age of HCV-infected patients with cryoglobulinemia was higher than age of HCV-infected patients without cryoglobulinemia. Microscopic examination of liver biopsy specimens revealed necroinflammatory activity slightly more prominent in patients with cryoglobulinemia. The most prominent inflammatory changes connected with abundant lymphoid aggregates in most of the examined portal tracts and piecemeal necroses were diagnosed in patients with several extrahepatic manifestations, such as cutaneous manifestations, nephrotic syndrome, polyneuropathy, and arthropathy. Liver fibrosis was similar in patients with and without cryoglobulinemia. CONCLUSIONS: The serum levels of all proinflammatory cytokines, especially IL-8, were significantly higher in the patients with cryoglobulinemia in comparison with the patients without cryoglobulinemia and healthy persons. All microscopic features did not correlate with the level of any investigated proinflammatory cytokines.

Composition of inflammatory infiltrate and its correlation with HBV/HCV antigen expression.

AIM: To study the composition of liver inflammatory infiltrate in biopsy material from patients chronically infected with hepatotropic viruses and to evaluate the correlation of inflammatory infiltrate with hepatitis B virus (HBV) and hepatitis C virus (HCV) viral antigen expression in chronic B and C hepatitis. METHODS: The phenotype of inflammatory cells was evaluated by the EnVision system, using a panel of monoclonal antibodies. HBV and HCV antigens were detected with the use of monoclonal anti-HBs, polyclonal anti-HBc and anti-HCV antibodies, respectively. RESULTS: The cellular composition of liver inflammatory infiltrate was similar in the patients with B and C hepatitis: approximately 50%-60% of cells were T helper lymphocytes. Approximately 25% were T cytotoxic lymphocytes; B lymphocytes comprised 15% of inflammatory infiltrate; other cells, including NK, totalled 10%. Expression of HLA antigens paralleled inflammatory activity. Portal lymphadenoplasia was found more often in hepatitis C (54.5%) than in hepatitis B (30.6%). Expression of HBcAg was found more often in chronic B hepatitis of moderate or severe activity. Overall inflammatory activity in HBV-infected cases did not correlate with the intensity of HBsAg expression in hepatocytes. Inflammatory infiltrates accompanied the focal expression of HCV antigens. A direct correlation between antigen expression and inflammatory reaction in situ was noted more often in hepatitis C than B. CONCLUSION: Irrespective of the etiology and activity of hepatitis, components of the inflammatory infiltrate in liver were similar. Overall inflammatory activity did not correlate with the expression of HBsAg and HCVAg; HBcAg expression, however, accompanied chronic hepatitis B of moderate and severe activity.

The presence and structure of circulating immune complexes in patients with prostate tumors.

BACKGROUND: The detection, measurement and characterization of circulating immune complexes (CICs) in tumor patients has shown both good and poor correlation with progression of the disease. The origin of the antigens making up the immune complexes in the sera of the cancer patients examined is unknown. They are perhaps antigens specific to the cancer's progress. MATERIAL/METHODS: The levels and molecular weights of circulating immune complexes were estimated in the sera of 48 patients with adenocarcinoma (grades G-1, G-2, and G-3) and 21 patients with benign prostate hyperplasia (BPH) by means of the polyethylene glycol (PEG) precipitation test. RESULTS: The results were compared with those of a group of 45 healthy blood donors. Elevated levels of CICs were observed in 66.7% of the patients with adenocarcinoma and in 38% of those with BPH. No seropositivity for CICs was observed in the control group. Increased seropositivity for CICs was observed in patients with the highness grade of adenocarcinoma: the proportion of seropositive patients in the group of patients with the G-1 phase of disease was 50%, with G-2 75%, and with G-3 71.3%. The molecular weights of the CIC proteins were determined by SDS/PAGE. The serum CICs of both BPH and prostate cancer patients consisted of proteins absent in the CICs of the sera of healthy persons. CONCLUSIONS: Determination and analysis of atypical proteins in CICs resulting from carcinogenesis may by useful in improving the diagnosis.

Relationship between clinical parameters and cytokine profiles in inflamed gingival tissue and serum samples from patients with chronic periodontitis.

OBJECTIVE: The purpose of the present study was to assess the relation between clinical parameters and concentrations of the key (IL-1beta, TNF-alpha, IL-2, IFN-gamma, IL-4, IL-10) cytokines, important in the initiation and progression of periodontal diseases, within inflamed gingival tissues and serum samples from patients with severe chronic periodontitis. MATERIAL AND METHODS: Twenty-five patients with severe chronic periodontitis, who had sites with probing depths (PD) > 5 mm, and 25 periodontally healthy persons were included in the study. Clinical examinations including PD, clinical attachment loss, plaque index, and bleeding index were performed before periodontal treatment. Gingival tissue biopsies were collected from one active site of each patient and from healthy individuals, and blood samples were withdrawn on the day of tissue biopsy. The concentrations of cytokines were determined by an enzyme-linked immunosorbent assay, and the relationship between their profiles in situ and in circulation with clinical parameters was analysed. RESULTS: The concentrations of IL-1beta, TNF-alpha, IL-2, IFN-gamma were, on average, significantly higher in serum samples and gingival tissue biopsies from periodontitis patients than in healthy controls. However, serum samples from both groups showed high individual variability of cytokine profiles, and no association between cytokine concentrations and clinical parameters of periodontitis was found. On the contrary, the levels of IL-4 and IL-10 in both kinds of samples obtained from patients and controls were generally low or even undetectable, and remained, on average, on the same level. However, the frequency of IL-4 (88% positive samples) and IL-10 (72%) was much higher in healthy gingival tissues. High concentrations of TNF-alpha, IFN-gamma and IL-2 and, especially, a high ratio of IL-1beta/IL-10 and TNF-alpha/IL-4 found in tissue biopsies from periodontitis patients, strongly correlated with the severity of periodontitis. CONCLUSION: These results indicate that high variability of cytokine concentrations and low frequency of their detection in serum samples from periodontitis patients make these determinations useless for the detection of disease presence and/or its severity. In contrast, high absolute levels of IL-1beta, TNF-alpha, IL-2 and IFN-gamma and, especially their high ratios to IL-4 and IL-10 found in inflamed tissue biopsies, were closely associated with periodontal disease severity.

Tubulointerstitial nephritis with uveitis: clinico-pathological and immunological study.

A 10-year-old boy was evaluated for fever, weight loss, uveitis, normocytic, normochromic anemia, renal insufficiency, and hypergammaglobulinemia of 8 weeks' duration. Infectious and neoplastic causes of fever were excluded. A renal biopsy performed in the 4th week of disease revealed diffuse plasmocytic interstitial nephritis. No treatment was prescribed and the patient was transferred to another hospital. Because clinical symptoms and renal insufficiency were still present, in the 8th week of disease a second biopsy was performed, which showed lympho-monocytic interstitial nephritis. At the same time, phenotypic analysis of peripheral blood mononuclear cells was carried out, revealing a significantly decreased number of CD3(+), CD4(+), and CD3(+)/CD8(+) cells, increased non-T CD3(-)/CD8(+) and CD56(+) NK cells, and decreased "naïve" (CD45RA(+)/CD4(+)) and memory (CD45RO(+)/CD8(+)) T lymphocytes. A 6-month course of oral prednisone was prescribed. Clinical symptoms and laboratory findings quickly returned to normal values. After 13 days of corticosteroid therapy, a second phenotypic analysis of peripheral blood mononuclear cells was performed, which revealed normalization of CD3(+), CD4(+), and CD3(+)/CD8(+) cells as well as proportions of non-T CD8(+) and CD56(+) NK lymphocytes, "naïve" and memory cells. This case shows spontaneous evolution of tubulointerstitial infiltrates from plasmacytic to lympho-monocytic, profound disturbances of the immunological system, and the beneficial effect of corticosteroids on both the clinical course and immunological disturbances.

[Individual sensitivity of jejunal mucosa to small doses of gluten in coeliac disease ]. / Osobnicza wrazliwosc blony sluzowej jelita cienkiego w celiakii na mala dawke glutenu.

In coeliac patients the age of development of symptoms, clinical picture of the disease and complications depend on the dose of ingested gluten. The aim of the study was the evaluation of individual sensitivity to small doses of gluten in the group of 60 patients aged 2.65 to 17.92 (mean age 7.49) treated with gluten-free diet for at least 12 months due to coeliac disease diagnosed according to ESPGAN criteria (food allergy to gluten excluded). Gluten challenge with dose of 10 mg/kg body mass/day was controlled with serological tests (IgAEmA, IgAAGA, and IgGAGA antibodies) carried out every 3 to 6 months. Jejunal biopsy was performed before gluten challenge (normal mucosa), and after positive EmA/AGA antibodies tests to confirm diagnosis (flat mucosa). After 35 months of observation 53.7% of all patients presented of jejunal villious atrophy, and positive IgAEmA. In this group 3.7% presented symptoms after 3 months of gluten challenge, 5.5% after 6 months, 3.7% after 9 months, and 3.7% after 12 months. In some coeliac patients ingestion of small amounts of gluten (10 mg/kg/day) can lead to small intestinal villious atrophy.