Hepatitis E virus serological testing in kidney transplant recipients with elevated liver enzymes in 2007–2011 in southeastern France

Hepatitis E virus (HEV) is an emerging cause of acute and chronic hepatitis in Europe, particularly in solid organ transplant recipients. Anti-HEV IgG/IgM testing in kidney transplant recipients with liver biological disturbances indicated high HEV exposure in our geographical area and led to diagnose HEV infection in 6% of cases.     

Seroprevalence of hepatitis E virus infection in rural and urban populations, Tunisia

Hepatitis E virus (HEV) is one of the leading agents of acute hepatitis. This study investigated the prevalence and risk factors of HEV infection in the Tunisian adult general population, either in blood donors (n=687) or in patients hospitalized for acute hepatitis (n=202). The mode of transmission differed between these two populations: contact with animals and living in a rural habitat were the main risk factors for being in contact with HEV in asymptomatic blood donors, while HEV was contracted through contaminated water in symptomatic cases. HEV seroprevalence in adult blood donors in Tunisia was relatively low (5.4%) and increased with age.     

Trypanosoma cruzi inactivation in human platelet concentrates and plasma by a psoralen (amotosalen HCl) and long-wavelength UV

Trypanosoma cruzi, the protozoan pathogen that causes Chagas' disease, can be found in the blood of infected individuals for their entire life span. This presents a serious challenge in safeguarding blood products. Transmission of T. cruzi from blood products is a frequent occurrence in Latin America, where Chagas' disease is endemic. This study was designed to determine whether T. cruzi could be inactivated in human platelet concentrates and plasma by a photochemical treatment process with long-wavelength UV A light (UVA, 320 to 400 nm) plus the psoralen amotosalen HCl (Cerus Corporation). Units of platelet concentrates (300 ml) and plasma (300 ml) were intentionally contaminated with approximately 10(6) T. cruzi trypomastigotes, the T. cruzi form found in the bloodstream, per ml. The viability of T. cruzi after photochemical inactivation was determined by their ability to replicate in 3T3 fibroblasts. Controls, including treatment with 150 micro M amotosalen or 3 J/cm(2) UVA alone, did not lead to reduction of the viability of T. cruzi in plasma or platelet concentrates. However, treatment with 150 micro M amotosalen plus 3 J/cm(2) UVA inactivated T. cruzi to undetectable levels in plasma and platelet concentrates. This represented a >5.4-log reduction of T. cruzi in platelet concentrates and >5.0-log reduction of T. cruzi in plasma. We conclude that the amotosalen plus UVA photochemical inactivation technology is effective in inactivating high levels of protozoan pathogens, such as T. cruzi, in platelet concentrates and plasma, as has been previously shown for numerous viruses and bacteria.     

First human case of co-infection with two different subtypes of hepatitis e virus

Hepatitis E virus (HEV), the main etiologic agent of enterically transmitted acute hepatitis in developing countries, is now recognized as an emerging agent of autochthonous disease and chronic hepatitis E in immunocompromised patients in Europe where HEV infection is probably zoonotically acquired. We describe the first human case of acute HEV infection with two genotype 3 viruses in a French kidney transplant recipient, probably acquired through consumption of uncooked pig liver sausage (figatellu). The patient presented two viral sequences nearly identical to sequences recovered from figatelli. Autochthonous co-infections with different genotype 3 HEV strains can occur in our geographical area.     

The role of photochemical treatment with amotosalen and UV-A light in the prevention of transfusion-transmitted cytomegalovirus infections

Primary cytomegalovirus (CMV) infection is usually asymptomatic in immunocompetent patients but can cause serious life-threatening complications in immunocompromised CMV-seronegative patients, including patients receiving a bone marrow or peripheral blood stem cell transplant, recipients of some solid-organ transplants, and low-birth-weight neonates. Current recommendations for preventing transfusion-transmitted CMV (TT-CMV) infection in these patients include exclusive use of CMV-seronegative and/or leukoreduced cellular blood components (red blood cells and platelets) for transfusion. However, breakthrough cases of TT-CMV still occur. Despite improving the safety of blood components, testing remains a reactive approach to blood safety. In contrast, pathogen inactivation technologies offer a proactive approach with the potential to further improve blood safety. To reduce the risks associated with platelet transfusions, a photochemical treatment (PCT) process using a combination of the psoralen amotosalen HCl and long-wavelength UV light has been developed and introduced into clinical practice in Europe. PCT has been shown to result in greater than 5.9-log reductions in infectivity of human CMV in platelet concentrates and to prevent the transfusion transmission of murine CMV in a mouse transfusion model. Thus, PCT pathogen inactivation may play a role in further reducing the incidence of TT-CMV infection in patients who are at risk for serious CMV disease. Because PCT is a technology that targets nucleic acids, it also offers a proactive process for the inactivation of a broad range of viral, bacterial, and protozoan pathogens in addition to CMV.     

Functional characteristics of photochemically treated platelets

BACKGROUND: A photochemical treatment (PCT) process using the psoralen compound amotosalen HCL (S59) and long wavelength UVA light was developed for inactivation of infectious pathogens and WBCs. In this study the effect of PCT on functional characteristics of the platelets was evaluated in vitro. STUDY DESIGN AND METHODS: Platelet concentrates were treated photochemically using the experimental clinical processing system T-bag S59 Reduction Device (SRD) (n = 4) or the commercially available integral processing system Wafer SRD (n = 4) and compared with control platelet concentrates in plasma/PAS III alone (n = 4). The evaluation included variables with respect to the overall quality of the product (e.g., HSR, pH), the function (aggregation and activation tests), apoptosis (annexin V and caspase 3), and lysis. RESULTS: No differences were found in the product quality variables, in P-selectin expression, and the apoptosis variables. PCT using the T-bag SRD led to a significant decrease in aggregation capacity with collagen and thrombin and a significant increase in plasma LDH, whereas no differences for the Wafer SRD were found. CONCLUSION: PCT using the experimental T-bag SRD led to a significant decrease in platelet function. However, the commercially available Wafer SRD had only minor in vitro effects on the quality of the platelets.     

Effects of intercept pathogen inactivation on platelet function as analysed by free oscillation rheometry.

INTRODUCTION: The Intercept Blood System, using InterSol as additive solution, is used for inactivation of contaminating pathogens in PCs, thus reducing the risk for transfusion transmitted infection and making it possible to prolong the storage period. This study aimed at investigating the ability of Intercept treated platelets to induce clot formation, as measured by coagulation time using free oscillation rheometry (FOR), and to compare with that of platelets in concentrates with the additive solution T-Sol or plasma. METHODS: Seventy-four single-donor platelet units were diluted in InterSol (n=27) or T-Sol (n=47) to a mean plasma concentration of 38%. The Intercept treatment was performed by addition of amotosalen HCl to the InterSol PCs followed by UVA irradiation and treatment with a compound adsorption device (CAD). Forty-six units were collected and stored in 100% plasma for comparison. Clotting time was measured by FOR in fresh PCs (within 26h after collection) after stimulation by a platelet activator. Soluble P-selectin was analysed as a marker of platelet activation in the Intercept and T-Sol PCs. RESULTS: The clotting time was shorter for Intercept treated platelets compared to platelets in T-Sol and plasma (p<0.05). There was no difference in clotting time between T-Sol and plasma PCs. Soluble P-selectin was higher for Intercept platelets than platelets in T-Sol (p<0.05). CONCLUSIONS: The platelets treated with the Intercept procedure had good clot promoting capacity.     

Photochemical treatment of platelet concentrates with amotosalen hydrochloride and ultraviolet A light inactivates free and latent cytomegalovirus in a murine transfusion model

BACKGROUND: A photochemical treatment (PCT) process utilizing amotosalen hydrochloride and long wavelength UVA light has been developed to inactivate pathogens in PLTs. This study investigated the effects of amotosalen/UVA treatment on free and latent murine CMV (MCMV) in PLT preparations using a murine model of transfusion-transmitted CMV (TT-CMV). STUDY DESIGN AND METHODS: In a model of latent MCMV infection, "donor" mice received 1 x 10(6) plaque-forming units (PFUs) MCMV and were rested 14 days. Subsequently harvested, pooled, and washed WBCs were PCR positive for MCMV. Murine WBC doses of 1 x 10(4), 1 x 10(5), and 1 x 10(6) were added to human apheresis PLTs in 35 percent autologous plasma and 65 percent PLT AS (PAS). The WBC-PLT products were treated with 150 micro mol/L amotosalen and 0.6 J per cm2 UVA and transfused via tail vein injection into recipient mice. Recipients were killed on Day 14. Blood and spleens were collected and assayed for MCMV by PCR. In a parallel model of active infection with free virus, human PLT in 35 percent autologous plasma and 65 percent PAS were dosed with 1 x 10(5) and 1 x 10(6) PFUs of MCMV. All other procedures were as described above. RESULTS: In the absence of amotosalen/UVA-pretreatment, transfusion of PLT latently or actively infected with MCMV produced TT-CMV in a dose-dependent fashion. In contrast, all transfusion recipients of identical PLT preparations pretreated with amotosalen/UVA were uniformly PCR negative for MCMV (abrogation of TT-CMV; p < 0.05). CONCLUSIONS: PCT of PLT preparations with the specified doses of amotosalen hydrochloride and UVA light prevents transfusion transmission of free and latent MCMV in a murine model. These results suggest that PCT of human PLTs with amotosalen/UVA should also effectively abrogate TT-CMV in the clinical setting.     

实时荧光逆转录PCR检测戊型肝炎病毒RNA的临床应用

目的 探讨实时荧光逆转录(RT)-PCR方法检测急性戊型肝炎患者血清戊型肝炎病毒(HEV)RNA的意义.方法 采用实时荧光RT-PCR方法,对HEV ORF3基因保守区进行扩增和检测.收集北京佑安医院住院和门诊434例急性戊型肝炎患者血清;甲型肝炎患者40例、乙型肝炎患者100例、健康献血员110名作为对照组;提取HEV RNA进行荧光RT-PCR检测.结果 434例急性戊型肝炎患者血清中232例(53.5%)为HEV RNA阳性.对照组血清HEV RNA检测均为阴性.与血清抗-HEV IgM检测比较,434例急性戊型肝炎患者HEV RNA检测的总符合率为67.1%,两种检测方法差异有统计学意义(Kappa=0.308,P=0.000).5例患者的首份血清检测为HEV RNA阳性,但抗-HEV IgM为阴性,系列追踪检测,相继出现抗-HEV IgM阳性.急性戊型肝炎患者的血清HEVRNA的检出多在发病的2～10 d内.结论 荧光RT-PCR方法有较高的特异性,应用实时荧光RT-PCR方法能对Ⅰ和Ⅳ戊型肝炎病毒感染的患者血清中的RNA进行定性检测.在临床使用可以提高对HEV早期诊断的水平.     

Leishmania inactivation in human pheresis platelets by a psoralen (amotosalen HCl) and long-wavelength ultraviolet irradiation

BACKGROUND: Leishmania spp. are protozoans that cause skin and visceral diseases. Leishmania are obligate intracellular parasites of mononuclear phagocytes and have been documented to be transmitted by blood transfusion. STUDY DESIGN AND METHODS: This study examines whether Leishmania can be inactivated in human platelet (PLT) concentrates by a photochemical treatment process that is applicable to blood bank use. Human PLT concentrates were contaminated with Leishmania mexicana metacyclic promastigotes or mouse-derived Leishmania major amastigotes and were exposed to long-wavelength ultraviolet (UV) A light (320-400 nm) plus the psoralen amotosalen HCl. RESULTS: Neither treatment with amotosalen nor UVA alone had an effect on Leishmania viability; however, treatment with 150 micromol per L amotosalen plus 3 J per cm(2) UVA inactivated both metacyclic promastigotes and amastigotes to undetectable levels, more than a 10,000-fold reduction in viability. CONCLUSIONS: This study demonstrates the effectiveness of photochemical treatment to inactivate Leishmania in PLT concentrates intended for transfusion. Both metacylic promastigotes, which represent the infectious form from the sand fly vector, and amastigotes, which represent the form that grows in mononuclear phagocytes, were extremely susceptible to photochemical inactivation by this process. Thus, the photochemical treatment of PLT concentrates inactivates both forms of Leishmania that would be expected to circulate in blood products collected from infected donors.     

The epidemiology of hepatitis E virus infections in developed countries and among immunocompromised patients

Hepatitis E virus (HEV) is an important cause of acute hepatitis in humans worldwide, both as epidemic and sporadic disease. Since the virus was identified in 1983, epidemics have occurred regularly in many countries across South and Southeast Asia when seasonal floods have contaminated drinking water supplies and in Africa during humanitarian crises among refugee populations without access to clean water. In addition, sporadic cases and small clusters of HEV infections have been recognized throughout the world in developed countries over the past couple of decades. This review will focus on emerging evidence of HEV infection as an under-recognized pathogen in Europe, the USA and other industrialized countries. We will discuss some of the issues associated with the recognition, diagnosis and treatment of these sporadic cases. We will also summarize the recent literature on autochthonous HEV infection among populations in developed countries in industrialized Europe, the USA, Japan and other industrialized Asian countries. We will review recent reports of acute and chronic HEV infections among transplant recipients and other immunocompromised individuals including HIV/AIDS patients.     

Mutation in the Hepatitis E Virus Polymerase and Outcome of Ribavirin Therapy

Hepatitis E virus (HEV) can lead to chronic infection in solid-organ transplant patients. Ribavirin is efficient for treatment of chronically infected patients. Recently, the1634R mutation in the HEV polymerase has been associated with treatment failure. However, it is unclear if this mutation can be used as a prognostic marker of treatment outcome. We studied the prevalence of the 1634R mutation in the HEV polymerase of patients starting ribavirin therapy, the influence of the 1634R variants on the viral response, the frequency of the 1634R mutation in patients whose treatment failed, and its impact on ribavirin retreatment. We analyzed pretreatment samples from 63 solid-organ transplant patients with chronic hepatitis E using deep sequencing; 42 patients had a sustained virologic response (SVR), and 21 were non-SVR patients. We detected the 1634R variant by deep sequencing in 36.5% (23/63) of the patients (proportions, 1.3 to 100%). The 1634R variant was detected in 31.0% (13/42) of baseline plasma samples from patients with SVR and in 47.6% (10/21) in the other patients (P = 0.2). The presence of this mutation did not influence the initial decrease in viral RNA. Lastly, a second prolonged ribavirin treatment led to SVR in 70% of the patients who initially did not have SVR, despite the presence of the 1634R variant. We conclude that the presence of the 1634R variant at ribavirin initiation does not lead to absolute ribavirin resistance. Although its proportion increased in patients whose treatment failed, the presence of the 1634R variant did not compromise the response to a second ribavirin treatment.     

The epidemiology of hepatitis E virus infections in developed countries and among immunocompromised patients

Hepatitis E virus (HEV) is an important cause of acute hepatitis in humans worldwide, both as epidemic and sporadic disease. Since the virus was identified in 1983, epidemics have occurred regularly in many countries across South and Southeast Asia when seasonal floods have contaminated drinking water supplies and in Africa during humanitarian crises among refugee populations without access to clean water. In addition, sporadic cases and small clusters of HEV infections have been recognized throughout the world in developed countries over the past couple of decades. This review will focus on emerging evidence of HEV infection as an under-recognized pathogen in Europe, the USA and other industrialized countries. We will discuss some of the issues associated with the recognition, diagnosis and treatment of these sporadic cases. We will also summarize the recent literature on autochthonous HEV infection among populations in developed countries in industrialized Europe, the USA, Japan and other industrialized Asian countries. We will review recent reports of acute and chronic HEV infections among transplant recipients and other immunocompromised individuals including HIV/AIDS patients.     

Inactivation of parvovirus B19 in human platelet concentrates by treatment with amotosalen and ultraviolet A illumination

BACKGROUND: The human erythrovirus B19 (B19) is a small (18- to 26-nm) nonenveloped virus with a single-stranded DNA genome of 5.6 kb. B19 is clinically significant and is also generally resistant to pathogen inactivation methods. Photochemical treatment (PCT) with amotosalen and ultraviolet A (UVA) inactivates viruses, bacteria, and protozoa in platelets (PLTs) and plasma prepared for transfusion. In this study, the capacity of PCT to inactivate B19 in human PLT concentrates was evaluated. STUDY DESIGN AND METHODS: B19 inactivation was measured by a novel enzyme-linked immunosorbent spot (ELISPOT) erythroid progenitor cell infectivity assay and by inhibition of long-range (up to 4.3 kb) polymerase chain reaction (PCR), under conditions where the whole coding region of the viral genome was amplified. B19-infected plasma was used to test whether incubation of amotosalen with virus before PCT enhanced inactivation compared to immediate PCT. RESULTS: Inactivation of up to 5.8 log of B19 as measured by the infectivity assay, or up to 6 logs as measured by PCR inhibition can be achieved under non-limiting conditions. Inactivation efficacy was found to increase with incubation prior to UVA illumination. Without incubation prior to illumination 2.1 +0.4 log was inactivated as determined by infectivity assay. When measured by PCR inhibition, inactivation varied inversely with amplicon size. When primers that spanned the entire coding region of the B19 genome were used, maximum inhibition of PCR amplification was demonstrated. CONCLUSION: Under defined conditions, PCT with amotosalen combined with UVA light can be used to inactivate B19, a clinically significant virus that can be transmitted through blood transfusion, and heretofore has been demonstrated to be refractory to inactivation.     

Ribavirin Inhibits In Vitro Hepatitis E Virus Replication through Depletion of Cellular GTP Pools and Is Moderately Synergistic with Alpha Interferon

Hepatitis E virus (HEV) is a common cause of acute hepatitis that results in high mortality in pregnant women and may establish chronic infections in immunocompromised patients. We demonstrate for the first time that alpha interferon (IFN-α) and ribavirin inhibit in vitro HEV replication in both a subgenomic replicon and an infectious culture system based on a genotype 3 strain. IFN-α showed a moderate but significant synergism with ribavirin. These findings corroborate the reported clinical effectiveness of both drugs. In addition, the antiviral activity of ribavirin against wild-type genotype 1, 2, and 3 strains was confirmed by immunofluorescence staining. Furthermore, the in vitro activity of ribavirin depends on depletion of intracellular GTP pools, which is evident from the facts that (i) other GTP-depleting agents (5-ethynyl-1-β-d-ribofuranosylimidazole-4-carboxamide [EICAR] and mycophenolic acid) inhibit viral replication, (ii) exogenously added guanosine reverses the antiviral effects, and (iii) a strong correlation (R² = 0.9998) exists between the antiviral activity and GTP depletion of ribavirin and other GTP-depleting agents.     

In vitro photochemical inactivation of cell-associated human T-cell leukemia virus Type I and II in human platelet concentrates and plasma by use of amotosalen

BACKGROUND: Human T-cell leukemia virus Types I and II (HTLV-I and HTLV-II), blood-borne retroviruses found worldwide, can cause leukemia, immunosuppression, and severe neurologic diseases. In most countries, HTLV-I and -II screening is not performed systematically for blood donations. A new photochemical treatment (PCT) with a synthetic psoralen was developed to inactivate most pathogens in platelet (PLT) concentrates or plasma and to improve the safety of blood donations. STUDY DESIGN AND METHODS: Cell-associated HTLV-I or -II (10(6)/mL) was inoculated in full-size fresh PLT concentrates or fresh frozen plasma and treated with 150 micromol per L amotosalen (S-59) and different doses of long-wavelength ultraviolet A (UVA) light. The residual viral titer in the treated samples was assessed by a cocultivation assay on indicator cells. RESULTS: The inactivation obtained at a 3.0 J per cm2 UVA dose was greater than 5.2 log foci-forming units (FFUs) per mL for HTLV-I and 4.6 log FFUs per mL for HTLV-II in presence of human PLT concentrates and greater than 4.5 log FFUs per mL for HTLV-I and 5.7 log FFUs per mL for HTLV-II in the presence of human plasma. The residual infectivity was very low and shown as the limit of detection of the cocultivation assay. CONCLUSION: In human plasma or PLT concentrates, the retroviruses HTLV-I and -II were strongly sensitive to the PCT with 150 micromol per L amotosalen (S-59) and a 3.0 J per cm2 UVA dose. This high efficiency for photoinactivation of these retroviruses opens a possibility of improving the safety of PLTs or plasma transfusion in the future.     

Management of chronic viral hepatitis in the hematological patient

Introduction: Infection with HBV and HCV represents a growing challenge in the management of patients with hematological malignancies. Recently, hepatitis E (HEV) was recognized as an endemic infection in developed countries and as an emerging health problem in immunocompromised patients.

Areas covered: We reviewed the current knowledge on the impact of chronic viral hepatitis in the hematological setting. Epidemiological features, screening strategies and indications for treatment and monitoring have been explored and commented.

Expert commentary: Knowing patient’s complete HBV serostatus is mandatory in order to choose between treatment, prophylaxis or a pre-emptive approach. Recent guidelines favor treatment with high barrier molecules in all patients with chronic HBV infection and long lasting prophylaxis with those with inactive or resolved one. With regard to HCV, the new direct-acting antiviral agents have been safely administered in the hematological setting. Their use as first-line single treatment in indolent lymphomas, and combined with chemotherapy in aggressive ones, should be considered. Due to the existing risk of chronic HEV infection in the immunocompromised, screening with serum HEV-RNA should be performed in case of signs and symptoms indicative of hepatitis. In the event of HEV infection, reduction of immunosuppression and, if not feasible or unsuccessful, ribavirin treatment should be prescribed.