Maciej Bura

Seroprevalence of anti-HEV IgG in 182 Polish patients.

INTRODUCTION Hepatitis E virus (HEV) infection is an emergent disease in developed countries. HEV seroprevalence in such areas significantly exceeds values expected when one considers infection with this virus only as a problem restricted to classical endemic regions. To date, no related data are available in Poland. In this study we aimed to obtain HEV seroprevalence data and compare them with similar data for hepatitis A virus (HAV) in Polish patients. MATERIAL/METHODS From February 1st, 2013, to October 15th, 2013, we performed anti-HEV IgG (anti-HEV) tests (EIAgen HEV IgG Kit; Adaltis, Milano, Italy) in 182 patients (101 men and 81 women; 61 patients were HIV-positive) of one center in Poland, aged 19-85 (47.2 ± 14.2 years). RESULTS We found a 15.9% seropositivity rate for anti-HEV (16.3% of the study population with an unequivocal test result) and 38.5% for anti-HAV. In 6 cases (3.4%), anti-HEV-positive persons had never travelled abroad. In contrast to HAV seroprevalence data, there was no significant difference in HEV seroprevalence between young adults (18-40 years) and older patients (p<0.0001 and p=0.0967, respectively). Anti-HEV were found in 21.3% of HIV-infected individuals. CONCLUSIONS HEV infection may occur in Poland. Anti-HAV seropositivity among Polish patients is significantly higher than anti-HEV. In contrast to HAV, HEV seroprevalence is similar in younger and older patients. The clinical course of HEV infection in Polish citizens seems to be largely asymptomatic. Polish HIV patients may be more commonly exposed to HEV than similar individuals from other countries.

Contribution of genes for killer cell immunoglobulin-like receptors (KIR) to the susceptibility to chronic hepatitis C virus infection and to viremia.

BACKGROUND Natural killer (NK) cells are an important element of innate immunity against viruses, although their numbers decrease in the liver during chronic HCV infection. NK cells express a large panel of inhibitory and activating receptors. The most polymorphic of these are killer cell immunoglobulin-like receptors (KIRs) which are encoded by multiple genes that may be present or absent in given individuals depending on their genotype. This variability results in differential susceptibility to viral infections and diseases, including HCV infection and its consequences. AIMS AND METHODS The aim of this study was to test whether chronical infection with HCV and the viremia levels are associated with any KIR gene in the Polish population. We typed 301 chronically HCV-infected patients and 425 non-infected healthy individuals for the presence or absence of KIR genes and their ligands, HLA-C C1 and C2 groups as well as HLA-B and HLA-A Bw4-positive alleles. RESULTS We found that males, but not females, possessing KIR2DS2 and KIR2DL2 genes had a 1.7 higher probability to become chronically HCV-infected than males negative for these genes (p=0.0213). In accord with this, centromeric B region, containing KIR2DS2 and KIR2DL2 genes, was also associated with chronic HCV infection in males. In addition, patients of both genders possessing KIR2DS3 but not KIR2DS5 gene exhibited, on average, 2.6 lower level of viremia than HCV-infected individuals with other genotypes (p=0.00282). This was evident in those infected at a young age. KIR2DS3-positive patients also had lower mean levels of bilirubin than KIR2DS3-negative ones (p=0.02862). CONCLUSION Our results suggest a contribution of the KIR2DS2 and KIR2DL2 genes (cenB haplotype) to the susceptibility to chronic HCV infection, and an association of the KIR2DS3 gene in the absence of KIR2DS5 with low viremia levels.

Hepatitis E virus IgG seroprevalence in HIV patients and blood donors, west-central Poland

OBJECTIVE To assess hepatitis E virus (HEV) seroprevalence in HIV patients and blood donors from one region in Poland. METHODS A group of 490 persons (244 HIV patients and 246 blood donors) aged 18-55 years were examined using the anti-HEV IgG assay (Wantai Biological Pharmacy Enterprise, Beijing, China). An analysis of the association between certain factors and the presence of this HEV exposure marker was conducted in both groups. RESULTS An HEV seropositivity rate of 50.2% was found. There was no difference in HEV seroprevalence between blood donors (49.6%, 122/246) and HIV patients (50.8%, 124/244) (p=0.569). The anti-HEV IgG positivity rate increased with age as follows: 36.2% (59/163) in persons aged 18-30 years, 52.0% (92/177) in individuals aged 31-40 years and 63.3% (95/150) in those aged 41-55 years. HEV infection occurred in 56.4% (31/55) of people who had never travelled abroad. CONCLUSIONS Wielkopolska Region in west-central Poland is an area hyperendemic for HEV infection. In this part of Poland, the exposure of HIV-positive persons to this virus is not greater than that of healthy blood donors.

Hepatitis E virus antibodies in HIV-infected patients and blood donors from western Poland: A preliminary report

BACKGROUND Hepatitis E virus (HEV) infection is an emerging problem in industrialized countries, including Europe. Little data exists on HEV seroprevalence in Poland. OBJECTIVES The aim of this study was to assess the prevalence of anti-HEV IgG antibodies in Polish patients infected with the human immunodeficiency virus (HIV) and blood donors. MATERIAL AND METHODS Two hundred and ten individuals (n = 105 of HIV-infected patients and n = 105 of ageand sex-matched blood donors from the same area; 178 men and 32 women), aged 18-50 (median age: 38 years), were tested for the presence of anti-HEV IgG antibodies with the EUROIMMUN Anti-Hepatitis E Virus (HEV) ELISA (IgG) tests (Lübeck, Germany). Additionally, some simple clinical and laboratory data was collected. RESULTS The overall anti-HEV IgG prevalence was 2.4% (5/210). One HIV-positive patient (0.95%) and 4 blood donors (3.8%) were seropositive (p = 0.1745). All the HEV-exposed individuals were men with a history of travel abroad and no icteric disease in the past. CONCLUSIONS Exposure to HEV infection among Polish HIV patients and blood donors seems to be uncommon. Data on this issue is scarce and conflicting for HIV-infected individuals. Further investigations applying different serological tests and concomitant HEV RNA testing are needed to reliably assess the risk and practical impact of HEV infection in Poland.

Results of antiviral treatment of patients with chronic hepatitis C: experience of Poznan centre.

INTRODUCTION Hepatitis C virus (HCV) infection in Poland affects approximately 750 thousand persons. The prevention of cirrhosis and hepatocellular carcinoma, of which approximately 20% of patients with chronic hepatitis C virus are at risk, aims at eradication of the virus by applying antiviral treatment with pegylated interferon alpha with ribavirin. MATERIAL/METHODS In this paper the results of the standard treatment of chronic hepatitis C in a population of 169 adult patients in whom it was started in the period of 01.01.2007-30.06.2008 are analyzed. Moreover, the influence of various clinical, biochemical and viral factors on achieving therapeutic success in the form of the sustained virological response (SVR) was studied. RESULTS In the group of 128 patients who received the full course of antiviral treatment, the SVR was achieved by 67.2% of patients (86 persons), whereas regarding all 169 patients who started the therapy, the sustained disappearance of viremia was found in 53.2% of patients (90 persons). Regarding 155 persons in whom the treatment was not interrupted for reasons others than virology, this value was 55.5%. For the sustained disappearance of viremia the following was favorable: genotype 3 virus, age under 40 years, body mass up to 75 kg, correct value of body mass index (BMI), low gamma-glutamyl transpeptidase (GGTP) activity before the treatment, minimum advancement of liver fibrosis in a liver biopsy (S1), complete early biochemical response (cEBR), and moreover, the achievement of negation of viremia after 12 weeks of the treatment in a group of patients infected with genotype 1 (complete early virological response, cEVR). These factors were strongly correlated with each other and that is why an analysis by the method of logistic multiple regression was impossible. Adverse reactions to the treatment and other health problems were the reasons for earlier discontinuation of the standard therapeutic scheme in 14 patients, whereby the lack of an SVR occurred in 10 of them (71.5% which is 5.9% of the studied population).

Viral Hepatitis A in 108 Adult Patients During an Eight-Year Observation at a Single Center in Poland

BACKGROUND Hepatitis A is related to significant morbidity and occasional mortality. Based on data from the Polish National Institute of Hygiene, from 2000 to 2013 a mean of 213 hepatitis A cases were reported yearly. OBJECTIVES The aim of the study was to assess selected data in adults hospitalized for symptomatic hepatitis A during an eight-year period in a single center in the Wielkopolska Region of Poland. MATERIAL AND METHODS All the hepatitis A patients hospitalized in the center from 2005 to 2013 were analyzed retrospectively. Data were extracted from the medical records of these individuals. The disease was confirmed by anti-HAV IgM testing. RESULTS In total, 108 patients (71 men and 37 women), aged 18-65 years, were identified. All but 1 patient recovered (99.1%) and in 6 cases (5.6%) a relapse occurred. Risk factors for hepatitis A were identified in 56 patients (52%), with travel abroad being the most common one (32 patients); 19 cases were secondary and 5 patients were men who have sex with men. One hepatitis A outbreak was noted in the region during the study period. Acalculous cholecystitis was found in 33.3% of the patients who underwent abdominal ultrasound. This tended to be more common among older individuals (47.8% in patients over 40 vs. 22.6% in patients aged 18-40, p=0.0521). Patients with this finding had significantly higher mean peak ALT in comparison to those with no gallbladder abnormalities. CONCLUSIONS Although hepatitis A in adults is typically a benign, self-limited disease, it can occasionally have a fatal course. In a significant proportion of patients with an evident risk factor for hepatitis A, the possibility of active prophylaxis was not used. Hepatitis A should be regarded as a sexually transmitted infection. Acalculous cholecystitis is a frequent finding among adults with symptomatic hepatitis A.

IFN-λ4 gene polymorphisms, circulating IFN-λ3, and clinical variables in hemodialysis patients exposed to hepatitis E virus

Introduction Factors associated with hepatitis E virus (HEV) infection are rarely recognized in patients on renal replacement therapy (RRT), and the results of studies are inconsistent. Objectives We aimed to search for determinants of HEV seroprevalence among polymorphisms of the interferon‑λ4 gene (IFNL4) associated with seroclearance of hepatotropic viruses (IFNL4 rs12979860, rs8099917 near IFNL4), circulating interferon λ3 (IFN‑λ3), and clinical variables of patients treated with hemodialysis (HD) in a HEV‑endemic region. Patients and methods The study was carried out in 90 HD patients. HEV open reading frame 2 antigen (HEV Ag), immunoglobulin M and G antibodies to HEV (anti‑HEV IgM and anti‑HEV IgG, respectively) and IFN‑λ3 were tested, and IFNL4 polymorphic variants (rs8099917, rs12979860) were genotyped. Survival analysis was conducted concerning anti‑HEV IgG positivity. Results In the study group, there were 37.8% anti‑HEV IgG‑positive subjects. None was HEV Ag or anti‑HEV IgM positive. HD modalities utilizing high‑flux dialyzers (adjusted odds ratio [OR], 3.586; 95% confidence interval [CI], 1.142-11.263; P = 0.03) as well as major homozygosity in rs8099917 (adjusted OR, 4.933; 95% CI, 1.516-16.054; P = 0.008) and rs12979860 (adjusted OR, 3.537; 95% CI, 1.136-11.014, P = 0.03), but not circulating IFN‑λ3 levels, were positive determinants of anti‑HEV IgG positivity. Liver enzyme activities and C‑reactive protein levels tested as response variables to HEV exposure, as well as survival probability, were not different between patients stratified by anti‑HEV IgG positivity. Conclusions Among HD patients, IFNL4 polymorphisms and treatment with high‑flux HD are explanatory variables for isolated anti‑HEV IgG positivity indicating spontaneous HEV resolution.

Comparative Seroprevalence of Hepatitis A And E Viruses in Blood Donors from Wielkopolska Region, West-Central Poland

The objective of the present study was to investigate the seroprevalence of HAV and HEV in Polish blood donors (BDs). One hundred and ten randomly selected healthy BDs, living in Wielkopolska Region were tested for anti-HAV IgG and anti-HEV IgG with commercial assays. The seroprevalence of anti-HAV was 11.8%; anti-HEV were detected in 60.9% of BDs (p < 0.0001). Consumption of risky food was more common in anti-HEV-positive BDs (59.1% vs. 33.3%; p = 0.01). Twelve out of 20 BDs (60%) with no history of travel abroad were exposed to HEV. Wielkopolska Region, Poland should be regarded as a new HEV infection-hyperendemic area in Europe.

HEV seroprevalence can significantly change after re-assessment

Dear Editor, We read with interest the paper recently published in the Journal of Medical Virology by Baptista-González et al discussing the prevalence of anti-HEV IgG antibodies in persons living in Mexico City including women with a high-risk pregnancy and healthy blood donors (BDs). The Authors reported very low overall HEV seroprevalence (2.34%, 10/428). They found no seropositive blood donor (out of 110 tested) and only one pregnant women with reactivity to HEV out of 127 tested. Data on the seroprevalence of HEV infection among pregnant women in non-classic endemic countries (ie, areas other than Asia or Africa) are scarce. Therefore, every new report discussing this issue is valuable. However, we think that the level of exposure to HEV found by Baptista-González et al can be significantly underestimated. A few HEV seroprevalence studies used the Euroimmun assay. In a preliminary assessment of HEV IgG seroprevalence in Greater Poland region (central Europe, west-central Poland) we found antiHEV antibodies in 2.4% (5/210) of ageand sex-matched BDs and HIV patients with the use of the same version of the Euroimmun assay as Baptista-González et al. Finally, we tested 306 individuals (153 healthy BDs and 153 HIV-infected patients) aged 18-55 for the presence of anti-HEV IgG antibodies. Two assays were used, that is, the Euroimmun assay (Euroimmun Medizinische Labordiagnostika AG, Lübeck, Germany) in all BDs and in 104 patients and the Wantai assay (Wantai Biological Pharmacy Enterprise, Beijing, China) in all study participants. All tests were performed and interpreted according to the recommendations of both manufacturers (borderline results were considered negative). Additionally, for the Euroimmun assay, apart from using the original cut-off value (OCO; results were considered positive when the quantitative analysis found anti-HEV IgG antibodies to be ≥2.2 IU/mL) a lowered cut-off (LCO) value (ie, 0.8 IU/mL) was also applied, as suggested by Norder et al. Of note, this suggestion was partially adopted by the manufacturer for the most current quantitative version of this assay (presently, Euroimmun recommends interpreting results as follows: <0.8 IU/mL − negative; 0.8 IU/mL to <1.1 IU/mL − borderline; ≥1.1 IU/mL − positive). The analyzed groupswere comparedwith the test for proportions. All tests were considered significant at P < 0.05. The analysis was carried out using statistical package Statistica PL12 (StatSoft, Inc). All BDs and patients gave their written informed consent. The study was approved by the local Bioethics Committee. Anti-HEV kits were funded by Poznan University of Medical Sciences (number of funds: 502-01-02205314-04519) and Regional Blood Center in Poznan. The results of anti-HEV IgG prevalence assessment are shown in Table 1. First, these results clearly shows that HEV seroprevalence can significantly change after re-assessment with a validated assay (or an updated version of the same assay). It is a well-known fact. However, Baptista-González et al did not mention this issue as a potential limitation of their study. Low sensitivity of the older version of the Euroimmun assay in comparison to assays of other manufacturers was also reported by Norder et al and Avellon et al. Moreover, in previous studies from Mexico, the presence of anti-HEV IgG antibodies was found in 0.4-40.7% of the tested persons. HEV seroprevalence varied depending on age, region of Mexico, a studied population and, of course, an assay used. It should be stressed that our knowledge on human HEV seroprevalence in Mexico is based on studies using assays of unknown performance (no comparative studies, no validation in HEV RNA-positive patients). However, a recent paper has shown that the exposure of Mexican domestic pigs to the hepevirus in question (tested with the Wantai assay) is very common. Due to zoonotic spread of HEV it supports the hypothesis that HEV infection can be significantly more common in inhabitants of Mexico City than found by BaptistaGonzález et al. Second, after matching this study participants for age and sex the use of more sensitive assays revealed a trend toward higher HEV seroprevalence in BDs versus HIV patients. It can be result of a weaker humoral response in the latter group. Third, these results suggest that exposure to HEV in west-central Poland is unusually high. It can be due to the fact that Poland is among 10 top producers of pork in the world (www.fao.org/faostat/) and that its consumption in this country is significant. Moreover, environmental contamination with porcine excreta could influence the level of exposure to HEV in the region. It should also be mentioned that the Wantai assay is a highly sensitive diagnostic tool. Although some falsepositive results of anti-HEV IgG testing are possible there is much evidence that its specificity is acceptably good. Last, it can also be hypothesized that an undefined but locally prevalent HEV-like

Exposure to hepatitis E virus, hepatitis A virus and Borrelia spp. infections in forest rangers from a single forest district in western Poland

BACKGROUND Hepatitis E virus (HEV) infection is an emerging problem in developed countries. At least 2 zoonotic genotypes of the virus (HEV-3 and HEV-4) infect human beings. There are some data suggesting that forest rangers (FRs) can be at a higher risk of contact with HEV. OBJECTIVES The aim of this study was to assess the prevalence of HEV exposure markers in FRs from a single forest district in Greater Poland in relation to anti-HAV (hepatitis A virus) IgG, and anti-Borrelia spp. IgM and IgG antibodies. MATERIAL AND METHODS In total, 138 participants (48 FRs and 90 blood donors - BDs) were tested for anti-HEV IgM and IgG (EUROIMMUN Medizinische Labordiagnostika AG, Luebeck, Germany) and 96 individuals (48 FRs and 48 BDs) were tested for anti-HAV IgG (ARCHITECT immunoassays, Abbott Laboratories, Wiesbaden, Germany); anti-Borrelia IgM and IgG (EUROIMMUN kits) were assessed in FRs only. RESULTS Anti-HEV markers were detected in 3 participants (2.2%; IgM in 1 FR, IgG in 2 BDs), less frequently than anti-HAV (16 out of 96 individuals, about 17%; FRs 19% vs BDs 15%) or anti-Borrelia antibodies (18 out of 48 individuals, 37.5%) (p < 0.0001 for both). Older study participants (≥45 years of age) were more frequently HAV-seropositive (29% vs 4% of the younger individuals; p = 0.0012). CONCLUSIONS We failed to unequivocally prove HEV exposure in FRs. The HAV seroprevalence in this study paralleled the situation in the general population. Exposure to Borrelia spp. in FRs was common.