Lack of evidence for the transmission of hepatitis E virus by coagulation factor concentrates based on seroprevalence data.

BACKGROUND: Whether hepatitis E virus (HEV) infection can be transmitted by coagulation factor concentrates remains unclear. OBJECTIVES: The HEV seroprevalence in blood donors and recipients of coagulation factor concentrates was compared to obtain evidence of whether a transmission of HEV by coagulation factor concentrates could occur. METHODS: Archived samples from whole blood donors and patients who had received coagulation factor concentrates were investigated for the presence of anti-HEV IgG by ELISA. Western blotting was used to confirm the positive samples that showed reactivity in the ELISA. RESULTS: Of 357 blood donors, 68 (19%) presented IgG antibodies against HEV. Two of 92 patients who had received coagulation factor concentrates (2·2%) and 1 of the 69 patients who had received plasma-derived products (1·5%) tested positive for anti-HEV IgG. The seroprevalence of HEV in the patient group was significantly lower (P = 0·038) than that in the donor group. The two positive patients were a 72-year-old man treated with plasma-derived products and a 5-year-old girl treated with a recombinant coagulation factor concentrate. CONCLUSION: HEV seroprevalence was significantly higher in the blood donors than in the patients with a history of coagulation factor concentrate administration. In one of two patients with detectable anti-HEV IgG antibodies, the coagulation factor concentrate was not the probable source of infection. Our data suggest that HEV is efficiently inactivated during the manufacturing process of coagulation factor concentrates. Thus, testing for the presence of HEV RNA in plasma donated for the preparation of coagulation factor concentrates may not be necessary.

Look-back study on recipients of Parvovirus B19 (B19V) DNA-positive blood components.

BACKGROUND AND OBJECTIVES: To assess the relevance of Parvovirus B19 (B19V) DNA at low to intermediate concentrations in blood donors for the recipients of their blood components. MATERIAL AND METHODS: We studied recipients of B19V DNA-positive blood components [red blood cell concentrates (RBCs), pooled platelet concentrates and fresh frozen plasma]. This included archived pretransfusion samples as well as follow-up samples investigated by ELISA or NAT and genome sequence analysis. RESULTS: In 132 out of 424 recipients, we could detect no anti-B19V IgG before transfusion. In 67 out of 132 sero-negative recipients, a follow-up sample was available. Sixty-five of these received blood components from donors with <10(4) IU B19V DNA/ml plasma and had no evidence of transfusion-transmitted (TT)-B19V infection. Homology in genome sequences in donor and recipient provided evidence for a TT-B19V infection in two recipients. Both patients received RBC containing 3.4 × 10(6) and 1.8 × 10(4) IU B19V DNA/ml plasma, respectively. The anti-B19V IgG titres in the donors were 2 and 76 IU/ml plasma, respectively. The antibodies in the second donor were directed against capsid proteins and are thus considered as potential neutralizing antibodies. CONCLUSIONS: TT-B19V infections through blood components with low (<10(4) IU/ml plasma) B19V DNA concentrations did not occur in our study. One of the TT-B19V infections occurred from RBC with intermediate B19V DNA concentration despite the presence of potential neutralizing antibodies in the donor, but its clinical significance was low.

Persistence of Parvovirus B19 (B19V) DNA and humoral immune response in B19V-infected blood donors.

BACKGROUND AND OBJECTIVES: Parvovirus B19 (B19V) DNA seems to persist in the plasma of B19V-infected blood donors. The relevance of this for recipients of single-donor blood components is yet unclear. MATERIAL AND METHODS: We studied serial archive and follow-up samples from 75 B19V-infected blood donors to obtain more data about the duration and degree of viraemia and the presence of IgG and IgM anti-B19V. IgG antibodies were further characterized by Western blot analysis in 29 donors. RESULTS: In 411 B19V DNA-positive samples collected, we found high concentrations (>10(6) IU B19V DNA/ml plasma) in five. B19V DNA persisted for a mean of 21·5 months (range: 2·3-52·4; 95% confidence interval, 19·1-23·9 months) in all donors. Only 15 such samples had either no or low-titre IgG anti-B19V. IgG antibodies were predominantly directed against epitopes on the minor capsid protein VP1, thus probably of neutralizing type with high avidity. IgM anti-B19V was detectable in 9/13 samples with high DNA concentrations. CONCLUSIONS: The vast majority of single-donor blood components with detectable B19V DNA are probably not infectious for their recipients because DNA is at only low levels and the donors also have potentially neutralizing antibodies with high avidity. Anti-B19V IgM testing does not identify every donation with high B19V DNA concentrations, but, in addition to B19V NAT testing, donors with persistent IgG anti-B19V might be considered 'B19V-safe' for single-donor blood components.

Comparison of the two fully automated anti-HCMV IgG assays: Abbott Architect CMV IgG assay and Biotest anti-HCMV recombinant IgG ELISA.

OBJECTIVE AND BACKGROUND: To assess the performance characteristics of two fully automated human cytomegalovirus (HCMV) antibody tests. MATERIALS AND METHODS: Samples from negatively or not pre-screened blood donors were tested by the Biotest anti-HCMV recombinant IgG enzyme-linked immunosorbent assay (ELISA) in comparison to the Abbott Architect CMV IgG assay [chemiluminescent microparticle immunoassay (CMIA)]. For clarification, samples with discordant results between both assays were subjected to supplemental testing for anti-HCMV IgG, IgM and HCMV DNA in plasma. RESULTS: From 4938 samples tested, 362 delivered positive results in both assays (7.3%). 91 (1.8%) samples were discordant. Of 43 (two not further tested) samples positive only by ELISA, 41 were false positive, one true positive and one indeterminate. Of 45 (one not further tested) samples positive only by CMIA, 20 were false positive, 9 indeterminate and 16 true positive. Anti-HCMV IgM and HCMV DNA testing from the plasma were negative in indeterminate samples. Considering the results of supplemental testing, the CMIA achieved altogether better results concerning resolved sensitivity, resolved specificity as well as negative predictive value. Both assays had an inferior positive predictive value, with a better result for CMIA. CONCLUSION: Overall, the performance characteristics of the CMIA were better than those of the ELISA. Owing to the inferior positive predictive value, positive test results require confirmation if blood products from donors with remote HCMV infection should be administered.

Pancreatic carcinoma cell lines reflect frequency and variability of cancer stem cell markers in clinical tissue.

BACKGROUND: Pancreatic cancer is one of the most deadly malignancies with insufficient therapeutic options and poor outcome. Cancer stem cells (CSCs) are thought to be responsible for progression and therapy resistance. We investigated the potential of pancreatic cell lines for CSC research by analyzing to what extent they contain CSC populations and how representative these are compared to clinical tissue. METHODS: Six pancreatic cancer cell lines were analyzed by flow cytometry for CD326, CD133, CD44, CD24, CXCR4 and ABCG2. Subsequently, 70 primary pancreatic tissues were evaluated for CD326, CD133 and CD44 by immunohistochemistry. RESULTS: All the cell lines but one showed a stable expression pattern throughout biological replicates. Marker expression in clinical tissue of CD44 distinguished normal patients from pancreatic carcinoma patients with a sensitivity of 50% at 80% specificity and metastasized from nonmetastasized carcinomas with 69% sensitivity at 100% specificity. CONCLUSIONS: Our results indicate a link between elevated CD44 expression, malignancy and metastasis of pancreatic tissue. Furthermore, individual pancreatic cell lines show a substantial amount of cells with CSC properties which is comparable with interpatient variability detected in primary tissue. These pancreatic cancer cell lines could thus serve for urgently needed pharmacological CSC in vitro research.

Preanalytical stability of HIV-1 and HCV RNA: impact of storage and plasma separation from cells on blood donation testing by NAT.

BACKGROUND: The aim of this study was to evaluate the optimal preanalytical conditions prior to nucleic acid amplification technology (NAT) for human immunodeficiency virus-1 (HIV-1) or Hepatitis C virus (HCV) RNA in pools of 96 plasma specimens with regard to storage temperature, time and plasma separation in a blood donation environment. STUDY DESIGN AND METHODS: Changes in viral nucleic acid concentration of HIV-1 and HCV were observed for 5 days according to the Paul-Ehrlich-Institute's (PEI) guidelines that demand 95%-detection limit of at least 10 000 IU mL(-1) for HIV-1 RNA and 5000 IU mL(-1) for HCV RNA within a single donor blood specimen. Ninety-five per cent detection limits of HIV-1 RNA over 3 days after storage at either 5 or 21 °C were evaluated by using standardised HIV-1 RNA-positive plasma. RESULTS: HCV RNA in whole blood samples proved to be more stable than HIV-1 RNA. Whole blood storage at 21 °C was shown to decrease the detectability of HIV-1 RNA even after only 18 h. Plasma samples once used for NAT at time 18 h did not alter viral stability up to 48 h after donation. Ninety-five per cent detection limits of HIV-1 RNA were securely below 10 000 IU mL(-1) for 24 h after whole blood storage at 5 °C. CONCLUSIONS: These results may lead to a discussion around the most suitable preanalytical conditions in blood donation environments. Contrary to the current PEI guidelines that allow storage of whole blood specimens up to 18 h at 21 °C, these results suggest that immediate storage in a 5 °C container after blood donation is more suitable and would permit storage of whole blood up to 24 h prior to the separation of plasma from cells.

The natural course of primary cytomegalovirus infection in blood donors.

BACKGROUND AND OBJECTIVES: As cytomegalovirus (CMV) DNA is frequently detectable in the plasma of recently infected sero-positive blood donors, information concerning primary CMV infection is important for the identification of possibly infectious donors. MATERIALS AND METHODS: Monitoring of 17 982 donors for CMV antibodies and DNA in plasma identified 14 subjects with ongoing primary CMV infection. Thirteen donors were interrogated for possible sources of infection and CMV-related symptoms, and monitored for CMV antigens, CMV DNA in plasma, leucocytes and urine, course of IgG and IgM antibodies as well as markers of systemic infection and parameters of organ function. RESULTS: CMV antigens and DNA were detectable in peripheral blood for up to 54 and 269 days respectively. Clearance of CMV DNA from blood correlated with clearance of IgM antibodies, development of IgG antibodies against the membrane glycoprotein gB and development of high avidity IgG antibodies. Eighty-five percent of subjects with primary CMV infection, but even 69% of matched controls reported possibly CMV-related symptoms. Sixty-two and 23%, respectively, had contact with possible sources of infection. One donor developed a febrile illness accompanied by increased levels of CMV DNA in peripheral blood 2 to 3 weeks after seroconversion. In other donors, neither markers of systemic infection nor parameters of organ function correlated with the course of CMV DNA and antigens. CONCLUSION: Potentially infectious donors can be identified by measuring CMV DNA, IgM antibodies or avidity of IgG antibodies. Alternatively, blood products donated during the first year after seroconversion should not be used for immunocompromised patients.

[Intestinal vasculitis and glomerulonephritis in hepatitis C- associated cryoglobulinemia]. / Intestinale Vaskulitis und Glomerulonephritis bei Hepatitis-C-assoziierter Kryoglobulinämie.

In a 53-year-old female patient with recurrent, sometimes bloody diarrhea, the long standing diagnosis of an ANA-negative lupus erythematosus with membranoproliferative glomerulonephritis, leucocytoclastic vasculitis and chronic hepatitis was ruled out and the diagnosis of a hepatitis C associated cryoglobulinaemia was established. The origin of the diarrhea was due to intestinal vasculitis as a result of cold food or beverages.

Expression of sCD23 in atopic and nonatopic blood donors: correlation with age, total serum IgE, and allergic symptoms.

Although structure, biologic activities, and expression of the low-affinity IgE receptor (FceRII, CD23) have been investigated, the diagnostic value for allergies of this molecule and its soluble circulating fragment (sCD23) remains unclear. Therefore, serum sCD23 levels were measured in 203 blood donors. They were divided into atopic and nonatopic subjects by allergy history, physical findings of allergic symptoms, and corresponding specific circulating IgE antibodies. The group consisting of nonatopic subjects was divided into four age categories in order to exclude age-dependent variations in the expression of the low-affinity IgE receptor. In our study population, sCD23 serum levels were not influenced by age. Furthermore, no significant differences, especially no decrease in serum sCD23 levels, between the four nonatopic age groups were detected. There was no significant increase of sCD23 serum levels in atopic subjects in comparison with nonatopic blood donors. In addition, no correlation between total IgE levels and sCD23 serum levels could be detected, in either the group of atopic donors or the group of nonatopics. Our data suggest that the circulating low-affinity IgE receptor does not appear to be an additional general marker for the diagnosis of allergies, as previously suggested.

[Sex and age-dependent gaps in tetanus immunization]. / Geschlechts- und altersabhängige Lücken im Tetanusimmunschutz.

To test tetanus immunity, tetanus antitoxin titres were measured in the serum of 692 subjects (354 males, 338 females), aged one day (newborns) to 92 years (mean age 29 years). Those aged 18 to 65 years were first-time blood donors, the remainder were healthy newborns, while the children and those over 65 years were patients without immune-compromising disease. An inadequate protection (titre < 0.1 IU/ml) was found in 107 (15.5%), of whom 75 (70%) were females. Women aged 20 years and above also had significantly lower average antitoxin titres than men (1.7 vs. 3.5 IU/ml); P < 0.0001). The inadequate immunization protection of many young women is reflected in the lack of protective antibodies in 10 of the 49 examined newborns. In addition, 18% of children aged between 1 and 15 years had inadequate immunity against tetanus. In the whole group the titre level decreased with age, while the proportion of unprotected persons increased. Apart from the obvious age and sex dependency of the demonstrated inadequacy of immunological protection against tetanus, attention should also be paid to the lack of protective antibodies in newborns and the marked gaps of immunity among children.

Immediate-type hypersensitivity reactions after platelet transfusion.

Non-febrile non-haemolytic transfusion reactions (NHTR) after platelet substitution vary from severe (anaphylactic shock) to mild (urticaria) forms. Although the symptoms of these immediate-type hypersensitivity reactions are well documented, little is known about their pathophysiology. We therefore examined sera from patients suffering from different forms of NHTR regarding total serum IgE, specific IgE antibodies and complement activity. Our results show that specific IgE antibodies in the recipients' sera were clearly associated with immediate-type hypersensitivity NHTR and conclude that atopic patients have a higher risk of suffering from these forms of transfusion reactions than non-atopic patients. Allergy diagnostic for patients who require platelet transfusions should be considered in case of multiple platelet substitution.

[Interferon alpha antibodies show no cross reactions with typical autoantibodies]. / Interferon-alpha-antikörper zeigen keine Kreuzreaktionen mit typischen Autoimmunantikörpern.

Patients treated with natural human interferon alpha develop anti-interferon antibodies (IFN-AB) only in very rare cases. By contrast, patients with autoimmune disorders are able to generate high-titered IFN-AB against endogenous interferon alpha. One explanation for the development of auto-IFN-AB could be cross-reactivity with typical autoimmune antigens. We investigated the cross-reactivity of 3 high-titered IgG IFN-AB of female autoimmune patients (aged 32, 36, 74 years; two severe cases of SLE, one case of autoimmune thyroiditis) as well as 25 low-titered natural IgM IFN-AB of healthy blood donors (aged 19-48 years). Typical autoimmune antigens including dsDNA, ENA, as well as natural interferon beta and recombinant interferon gamma are not able to inhibit binding of IFN-AB to interferon alpha in an ELISA test system. Preincubation of sera containing either dsDNA antibodies (dsDNA-AB) (24 patients), thyroid peroxidase (TPO-AB) (9 patients) or thyroglobulin (TG-AB) (12 patients) with interferon alpha resulted in no change in the respective autoantibody titer. These data suggest that there is no cross-reactivity between IFN-alpha-AB and dsDNA-AB, TPO-AB or TG-AB. Thus, an explanation for the occurrence of IFN-AB in autoimmune disorders cannot be found in a cross-reaction between interferon alpha with typical autoimmune antigens.