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Parvovirus B19 as a possible causative agent of fulminant liver failure and associated aplastic anemia.

We recently observed that more than one third of pediatric patients who presented with non-A, non-B fulminant liver failure (FLF) also developed aplastic anemia (AA) either before or shortly after liver transplantation. Factors involved in the suppression of bone marrow could be the same as those causing hepatic failure. We considered parvovirus B19 a candidate etiologic agent because of the known tropism of B19 for erythroid precursors. Archived liver and serum from six patients undergoing liver transplantation for non-A, non-B, non-C FLF with associated AA were analyzed for the presence of B19 DNA and anti-B19 serology. An age- and gender-matched control group (N = 44) was analyzed in parallel. B19 DNA studies and anti-B19 serology were performed in a blinded fashion. B19 serologies were performed by antibody capture enzyme-linked immunosorbent assay (ELISA). B19 DNA was detected after polymerase chain reaction (PCR) amplification of target B19 DNA sequences in liver and serum. Liver tissue showed evidence of B19 DNA in four of six (66%) patients with FLF and associated AA. Two of 4 patients with cryptogenic FLF but without AA had B19 DNA detected in the liver tissue. Of the 34 remaining controls, only 5 (15%) showed evidence of B19 DNA in liver tissue (66% vs. 15%, P = .016). B19 DNA was not detected in any of the test or control sera. This study provides evidence to support the role of parvovirus B19 in the development of FLF and associated AA.

Anemia, Aplastic↗

Parvovirus-based vectors for human gene therapy.

It is becoming increasingly clear that the parvovirus-based vectors may prove to be a useful alternative to the more commonly used retroviral vectors in human gene therapy. Specifically, the adeno-associated virus 2 (AAV), a human parvovirus, has gained particular attention in view of its nonpathogenic nature as well as its remarkable site-specificity of integration into the human chromosome. Using the recombinant AAV vector system, it is feasible to obtain high-efficiency transduction of slow- or non-cycling primary hematopoietic stem and progenitor cells, without the need for prestimulation with cytokines, which could potentially lead to differentiation of these cells before transplantation.

Antigens, CD↗

A nonlethal rat parvovirus infection suppresses rat T lymphocyte effector functions.

Inoculation of the UMass strain of rat virus (RV-UMass) into adult immunocompetent rats results in a prolonged subclinical infection that is resolved in 4 to 8 wk. Co-labeling studies, using in situ hybridization (ISH) and immunohistochemistry (IHC), confirmed that RV-UMass was lymphocytotropic and capable of infecting CD4+ and CD8+ T cells as well as B cells. ISH studies also revealed that virus replication was restricted in unstimulated cells but was productive in concanavalin A-stimulated lymphocytes. A corollary of productive infection of lymphocytes was the suppression of lymphocyte functions. Although RV-UMass did not appear to induce phenotypic changes during the course of infection, cells from infected rats had diminished proliferation and cytolytic responses. Both peripheral and mesenteric lymph node cells exhibited only partial recovery of their proliferative and cytolytic capacities one month after infection. Furthermore, RV-UMass-infected tissue culture maintained alloreactive CD4+ T cells in vitro, and a nonlethal infection of this T cell line inhibited Ag- and IL-2-induced proliferation. Because parvoviruses are widespread among laboratory rodents, these findings emphasize the importance of identifying and excluding parvovirus infection in rodents and in cultures of rat T lymphocytes.

Animals↗

[Parvovirus B19 infection: the general aspects and gestational problems].

Parvovirus B19 is a small, heat-stable, single-strained DNA virus (5.5 kb), with 23 nm icosahedral capsid discovered in 1975. Since its discovery, the virus has been shown to be a causative agent of erythema infectiosum (fifth disease). Under specific circumstances it can cause transient aplastic crisis, chronic anemia, arthritis and fetal death. Laboratory diagnosis of recent or past B19 infection usually relies on the demonstration of virus-specific IgM or IgG antibodies in patient's serum. The aim of this study was to evaluate the prevalence of Parvovirus B19 infection among 87 pregnant patients. Our study on anti-B19 antibody prevalence indicates that about 64% of pregnant women in our country lack specific antibodies and are at risk of B19 infection. One intrauterine death was encountered in a patient with specific IgM antibodies. The authors suggest the detection of IgG and IgM anti-B19 antibodies as a routine screening practice during pregnancy.

Adolescent↗

Isolation, identification, and plaque titration of parvovirus from Muscovy ducks in Japan.

Muscovy ducks (Cairina moschata) showed abnormal feathering, leg weakness, and high mortality. A virus was isolated from these ducks after several blind passages in embryonating Muscovy duck eggs. The isolate was resistant to chloroform, to pH 3.2, and to 65 C for 30 min. Electron microscopy showed that the isolate was an icosahedral and nonenveloped virus 20-22 nm in diameter. The isolate reacted with an antiserum against a goose parvovirus in agar gel precipitation tests. After 15 passages of the isolate in embryonating eggs, the isolate was adapted to Muscovy duck embryo fibroblasts. The adapted virus developed cytopathic effects and made clear plaques on sheets of the fibroblasts. When 5-iodo-2-deoxyuridine was added to the culture medium, virus growth was inhibited. From the data shown above, the isolate was identified as a goose parvovirus.

Animals↗

[Angiocentric lymphoma associated with anemia secondary to parvovirus B19 infection].

A 50-year-old woman was admitted to our hospital in May 1993 because of papules, cervical lymphadenopathy and interstitial pneumonia. Oxygen inhalation and pulse therapy of corticosteroid were started. A biopsy of the left inguinal lymphnode showed T zone enlargement and marked vascularization with polymorphous atypical lymphoblasts, consistent with angiocentric lymphoma (immunohistochemically T cell type). In situ hybridization with EBER-1 did not show association of EB virus. She was subsequently treated with ProMACE-CytaBOM successfully, but anemia had progressed before that treatment showing a reticulocyte count 0/1000, LDH 870 U/l, positive direct Coombs test. Bone marrow aspiration revealed red cell aplasia and the existence of giant pronormoblasts. The hemoglobin dropped to the level of 4.7 g/dl, requiring anabolic steroid and frequent blood transfusion. Parvovirus B19-specific antibodies were negative initially by western blot assay, but IgM antibody appeared after 35 days and IgG after 71 days. In August anemia improved following a reticulocyte burst and recovery of bone marrow erythroblasts. This patient is the first reported case of angiocentric lymphoma complicated with severe anemia, perhaps resulting from autoimmune hemolytic process and parvovirus B19 infection.

Anemia↗

Parvovirus B19 associated monoarthritis in a 5-year-old boy.

We describe the case of a 5-year-old boy who presented with parvovirus B19 associated arthritis of the left knee lasting for 6 weeks. Other features included flu-like symptoms, a mild "slapped cheek" sign, and a macular, erythematous, lace-like rash over the chest and limbs. The analysis of the synovial fluid showed a high white cell count with a predominance of polymorphonuclear cells. The characteristic features related to parvovirus B19 associated arthropathy in children are reviewed.

Arthritis, Infectious↗

[Arthropathy associated with infection by parvovirus B19. Description of 4 cases].

Four cases of human parvovirus infection in which the main clinical manifestation was a polyarthritis are described. Four females with ages ranging from 30 to 32 years presented with acute symmetrical polyarthralgias involving hands and knees. In addition, evidence of synovitis in the ankles and tenosynovitis of the fingers was found in two and three cases respectively. Half of the patients noticed an erythematous rash in the preceding days. Laboratory studies were normal in all cases. Antinuclear antibodies and rheumatoid factor were not detected in any case. All patients had significant levels of IgG and IgM antibodies to parvovirus B19 at the time of presentation and a rise in IgG and a fall in IgM levels were seen at two months. All cases cleared up within two weeks without treatment.

Adult↗

Cytopenia caused by parvovirus in an adult ALL patient.

A young woman in maintenance therapy for acute lymphoblastic leukemia in second complete remission developed fever and a skin rash associated with severe anemia, neutropenia and erythroblastopenia. A complete recovery was obtained in 4 weeks' time after red cell transfusion, i.v. immunoglobulin and withdrawal of the maintenance chemotherapy. Parvovirus B19 infection was demonstrated by detection of B19 DNA in the patient's serum using a dot-blot hybridization assay and a nested polymerase chain reaction. Serological tests were positive for anti-B19 IgG but not for IgM. Erythroblastopenia due to parvovirus infection has already been reported in ALL patients. B19 infection should be suspected in leukemic patients if unexplained cytopenia (mainly anemia) follows an acute febrile illness. Very sensitive methods are often needed to confirm the diagnosis, since routine serological tests may be unreliable in immunocompromised patients.

Adult↗

[Parvovirus B19 and pediatric pathology].

B 19 parvovirus is a widespread virus with primary infestation generally occurring in childhood through family and community outbreaks. Its most typical manifestation is transient erythroblastopenia with aplastic crisis, often profound, mostly affecting patients with chronic hemolytic anemia, and eventually patients with defective erythropoiesis (chronic hypoplastic anemia, iron deficiency anemia). In normal individuals the primary infestation is usually asymptomatic but may give transient hematological signs for few days: moderate reticulocytopenia, thrombopenia and neutropenia. Clinically two phases of the infection are described: 1.) a first phase of viremia of 2 to 3 days which may be accompanied by fever and myalgias; 2.) a second phase which may last for several weeks with dermatological signs, the most typical being erythema infectiosum, vasculitis, arthralgias or arthritis. In pregnant women, the primary infestation with B 19 parvovirus may lead to fetal anemia and hydrops fetalis with uneven outcomes: fetal death, chronic erythroblastopenia after birth, spontaneous resolution. Although the incidence of fetal infestation in non immunized pregnant women is still unknown, the question is raised of the recognition and protection of non immunized pregnant women at high risk of exposition to infested subjects. Long term persistence of the virus in the organism may be responsible for chronic manifestation, essentially but not exclusively in immunodeficient-patients: prolonged erythroblastopenia and chronic rheumatologic manifestations. It may be also responsible for cases of juvenile arthritis, thrombocytopenic purpura and chronic neutropenia of childhood. The diagnosis of the viral infestation is mainly based upon the detection of specific IgM, then IgG, antibodies by Elisa technique.

Child↗

Childhood idiopathic thrombocytopenic purpura: association with human parvovirus B19 infection.

PURPOSE: Infection with human parvovirus B19 is the most common cause of transient aplastic crisis in patients with chronic hemolytic anemia. Multiple reports of children with simultaneous B19 infection and thrombocytopenia as well as the known association between experimental B19 infection and thrombocytopenia prompted us to hypothesize that B19 may be associated with childhood idiopathic, or immune, thrombocytopenic purpura (ITP). Because there is a paucity of evidence regarding a viral etiology for ITP, we performed a comprehensive study to explore its possible relationship to B19 infection. PATIENTS AND METHODS: Thirty-five previously healthy children with ITP were studied prospectively. Bone marrow and peripheral blood were analyzed for B19 DNA using the polymerase chain reaction (PCR). Serum was analyzed for anti-B19 immunoglobulin (Ig) M and IgG antibodies using a B19 VP1 antigen-based enzyme-linked immunosorbent assay. Fourteen healthy children served as controls for peripheral blood PCR and serologic analyses. RESULTS: The presenting clinical and laboratory features of the study population were typical of classic ITP. Seventeen of the 35 patients (49%) had evidence of B19 DNA in the peripheral blood, bone marrow, or both. Six of 35 (17%) had anti-B19 IgM antibodies. Eight of 35 (23%) were anti-B19 IgG seropositive. The control group had no positive PCR or anti-B19 IgM specimens. CONCLUSIONS: Our results suggest that infection with human parvovirus B19 may be associated with childhood ITP. More investigation is warranted regarding the role of PCR methodology and serologic detection methods in defining B19 pathobiology as it relates to ITP.

Adolescent↗

Serologic response of pups to the low-passage, modified-live canine parvovirus-2 component in a combination vaccine.

Thirty pups from a general pet population were vaccinated for canine parvovirus-2 (CPV-2), using a low-passage, modified-live CPV-2 in a combination vaccine, every 3 weeks until the pups were between 11 and 18 weeks old. Canine parvovirus-2 antibody titers were measured by means of serum neutralization (SN) immediately before each vaccination and > or = 2 weeks after final vaccination. Thirteen pups that were initially seronegative (SN titer < 2 for CPV-2) developed protective titers (SN titer > or = 16) after 1 to 3 doses of the vaccine, administered when the pups were between 8 and 17 weeks old; 11 of the 13 developed protective titers after the first dose. Seventeen pups were initially seropositive for CPV-2 (SN titer > or = 2). Of these, 8 initially had titers < 16 and 3 others had titers that fell to < 16 during the study. Nine (82%) of these 11 pups developed protective titers. Results indicated that pups at high risk for CPV-2 disease should be vaccinated until 18 to 20 weeks old, even when the low-passage vaccine is used. Pups at lower risk of CPV-2 disease should be vaccinated periodically until 16 weeks old.

Animals↗

Erythema infectiosum and other parvovirus B19 infections.

The human parvovirus B19 has been identified as the causative agent in several diseases, including the benign childhood condition erythema infectiosum (fifth disease), a polyarthropathy that occurs in adults, aplastic crisis and hydrops fetalis. The virus has also been associated with numerous other hematologic, neurologic and rheumatic conditions. Except for the potential of neonatal death due to fetal hydrops, the prognosis of patients with parvovirus infection is generally good. The diagnosis of B19 infection may be established on clinical grounds but often requires confirmation with serum antibody testing. Treatment strategies may include supportive care, analgesic medications, transfusions with red blood cells or administration of intravenous immunoglobulin, depending on the clinical circumstances.

Animals↗

A modified plaque assay and infected cell hybridization assay for wild-type and recombinant LuIII autonomous parvovirus.

We describe a convenient infected cell hybridization assay for determining the infectious titer of a recombinant, replication-defective parvovirus. We previously generated recombinant derivatives of the autonomous parvovirus LuIII, transducing the luciferase reporter gene. Since luciferase expression is not readily detected at the single cell level, and since the recombinants cannot form plaques, we developed an alternative assay for infectious particles. Co-infection of NB324K cells with wild-type LuIII (multiplicity of infection ca. 5) and the recombinant virions allows amplification of the transducing DNA, which can be detected by hybridization with a probe for the reporter gene. Cell lysis and DNA transfer to a nylon membrane is performed in situ, in the culture dish, and hybridization is performed with a digoxigenin-labeled probe, using immunological detection. During this work we developed a conveniently modified plaque assay for wild-type LuIII that should be applicable to other lytic viruses. The modification employs a reduced volume of agarose overlay that is in turn overlaid with liquid medium, thus avoiding the need to maintain stocks of culture medium at higher than normal concentration.

Hybridization, Genetic↗

[Transient aplasia of the red progenitor cells manifest in a child with double heterozygote Hb SC carriership: the role of human parvovirus B19 (HPV-B19)].

Severe pure red cell anaemia due to B19 parvovirus infection is reported in a child with haemoglobin SC disease. B19 parvovirus infection was demonstrated by specific IgM. Although hypoplastic crisis occurring in SC children is usually less frequent and more benign than in sickle- cell anaemia, it necessitated several blood transfusions in our patient. The child recovered rapidly and was discharged after 12 days. This case again illustrates the role of HPV-B19 in the development of transient red cell aplasia occurring in patients with haemolytic anaemia. However, it is unusual since in the few reported children with HbSC disease and HPV-B19 infection, aregenerative anaemia remained moderate and did not generally require blood transfusions.

Anemia, Aplastic↗

[Cardiac involvement in fetal parvovirus B19 infection].

The review of 15 cases of cardiac involvement demonstrate the cardiac tropism of the human parvovirus B19. Ten of these cases were collected from fetuses during second trimester of maternal-fetal infections. In situ hybridisation detects viral DNA sequences in the nucleus of infected myoblasts. Myocarditis is the most frequent histological damage. Cardiac failure, secondary to myocarditis, may occur in the absence of fetal anaemia. When the fetus is deeply anaemic, like usually in cases of hydrops, damages of the cardiac tissues might hamper the reactional increase of cardiac output; therefore, they might account for the poor prognosis of parvovirus B19 fetal hydrops in the second trimester of pregnancy, despite transfusional therapy attempts in the third trimester.

Erythema Infectiosum↗

[Seroprevalence of IgG antibodies against parvovirus B19 in the population of the Czech Republic].

The human parvovirus B 19 was discovered in 1975 by Cossart in England. Later (1984) evidence was provided that this virus is the etiological agent of erythema infectiosum and hydrops foetalis, and in 1985 it was provided that it is also the etiological agent of some types of arthritis or arthropaties and vasculitis. The diagnosis of the disease caused by this agent is most frequently based on evidence of specific immunoglobulins. The epidemiological and clinical impact of parvoviral infections in the Czech Republic was not known so far. Examination of sera from 562 subjects aged 0-60 years assembled in 1992 in three Czech regions revealed in children, age 0 - 4 years 9.8%, during preschool and school age 27 - 35.7% and in age groups above 15 years a 53.3 - 57.7% seroprevalence of IgG parvovirus B 19, roughly equally distributed among both sexes. The more frequent prevalence of specific immunoglobulins was proved in small groups of female workers in nurseries (66.7%), nursery schools (91.7%) and in blood transfusion stations (77.8%). The seropositivity of the general female population of matched age groups, with the exception of women aged 20 - 24 years, was 53.86%.

Adolescent↗

[Rapid enzymatic test for diagnosis of parvovirus infections in dogs].

In this study 52 canine fecal samples were examined for the presence of canine parvovirus (CPV). The two different test systems used to confirm infection were electron microscopy as standard method and an enzyme-linked immunosorbent assay (ELISA). In comparison, the results conferred in 92.3%. The CPV ELISA had a sensitivity of 89.7% and a specificity of 100%. Therefore, it can be used as a quick, reliable method for diagnosis of canine parvovirus infection in veterinary practice.

Animals↗