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Further studies on thermal resistance of bovine parvovirus against moist and dry heat.

To supplement the results of thermal resistance of bovine parvovirus (Haden strain, BPV) published previously, we carried out assays at 60 degrees C (moist heat) to compare the thermal resistance of BPV with that of hepatitis B-virus (HBV). What we know about the resistance of HBV at a temperature of 60 degrees C is mainly based on data collected within the context of blood product pasteurization. The results suggest that at a temperature of 60 degrees C, BPV shows thermal resistance comparable to HBV. Thus, BPV--which is easier to handle--can be considered a good test virus to verify the efficacy of thermal disinfection techniques against HBV. BPV is very resistant against dry heat of 100 degrees C, the inactivation largely depending upon the residual moisture of the lyophilisate. Reducing the residual moisture from 2% to less than 1%, the exposure time has to be prolonged by ca. 2.5 times to achieve the same virucidal effect.

Animals↗

Human parvovirus B19 non-structural protein (NS1) induces apoptosis through mitochondria cell death pathway in COS-7 cells.

Human parvovirus B19 has been found in various tissues in addition to erythroid lineage cells, and non-structural protein (NS1) is reported to induce cytotoxicity and apoptosis in erythroid lineage cells, but the mechanism in non-permissive cells is still unclear. To address this issue, we have constructed the NS1 gene in a cytomegalovirus episomal vector, pEGFP-C1 and transfected it into monkey epithelial cells, COS-7. EGFP-NS1 expression in transfected cells was monitored and assessed by fluorescence microscopy, RT-PCR and Western blot. The flow cytometric analysis showed that the NS1-transfected cells were arrested at G1 phase by paclitaxel treatment and there was increased apoptosis. The expression of p53, an important molecule in apoptosis and cell cycle regulation, and its downstream cell cycle kinase inhibitors p16(INK4) and p21(WAF1/CIP1) were up-regulated in the NS1-transfected cells. Also, increased expression of the pro-apoptotic Bcl-2 members Bax, Bad and activation of caspase 3 and caspase 9, but not the activation of caspase 8 or Fas were detected in the NS1-transfected cells. p53-induced Bax expression and subsequent activation of caspase 9 is probably the apoptotic pathway in NS1-transfected cells since activation of the caspase 9 was suppressed by the p53 inhibitor and apoptosis was significantly inhibited by the caspase 9 inhibitor. Our results suggest that the cell death of the NS1-transfected cells is associated with mitochondria related apoptosis. These findings might provide alternative information for further study and characterization of B19 NS1 protein in B19 non-permissive cells.

Animals↗

Prevalence of parvovirus B19 DNA in bone marrow of patients with haematological disorders.

Patients with haematological disorders (n = 100) were examined for prevalence of parvovirus B19 DNA in the bone marrow and serum, irrespective of B19-related symptoms. B19 DNA was studied using 2 nested PCRs and the serum samples were further analysed with B19-specific IgG, IgM and avidity as well as seroreactivity against linear and conformational epitopes of the B19 VP2 antigen. The latter assays specify whether the IgG antibody response represents acute or past B19 infection. B19 DNA was detected in 4 of the 100 bone marrow samples, whereas all the serum samples were B19 DNA negative. None of the 4 B19 DNA positive patients had symptoms typical of B19 infection and serology showed past infection. Furthermore, 2 were still B19 DNA positive in bone marrow more than 1 y after the first sample indicating virus persistence. The seroprevalence for B19 IgG was 59% and 2 patients were B19 IgM positive. Thus, presence of B19 DNA in bone marrow from patients with haematological disorders is not a general finding in seropositive patients. B19 DNA can persist in bone marrow, but in our material this finding showed no clear correlation with symptomatic B19 infection.

Adult↗

Phylogenetic analysis of Hungarian goose parvovirus isolates and vaccine strains.

Polymerase chain reaction and sequencing were used to analyse goose parvovirus field isolates and vaccine strains. Two fragments of the genome were amplified. Fragment "A" represents a region of VP3 gene, while fragment "B" represents a region upstream of the VP3 gene, encompassing part of the VP1 gene. In the region of fragment "A" the deduced amino acid sequence of the strains was identical, therefore differentiation among strains could be done only at the nucleotide level, which resulted in the formation of three groups: Hungarian, West-European and Asian strains. In the region of fragment "B", separation of groups could be done by both nucleotide and deduced amino acid sequence level. The nucleotide sequences resulted in the same groups as for fragment "A" but with a different clustering pattern among the Hungarian strains. Within the "Hungarian" group most of the recent field isolates fell into one cluster, very closely related or identical to each other, indicating a very slow evolutionary change. The attenuated strains and field isolates from 1979/80 formed a separate cluster. When vaccine strains and field isolates were compared, two specific amino acid differences were found that can be considered as possible markers for vaccinal strains. Sequence analysis of fragment "B" seems to be a suitable method for differentiation of attenuated vaccine strains from virulent strains.

Amino Acid Sequence↗

Survival of a preterm neonate with late onset hydrops fetalis due to parvovirus B19 infection.

This paper presents a case of non-immune hydrops (NIH) which presented late at 28 weeks' gestation during an otherwise asymptomatic pregnancy. A severely hydropic female infant was born with significant cardiorespiratory compromise; however, aggressive resuscitation and postnatal treatment resulted in a favourable outcome. Parvovirus B19 DNA was subsequently detected in postnatal blood samples by nested polymerase chain reaction. This case emphasizes that survival is achievable even in those cases where intrauterine transfusion is not performed. It highlights the importance of prompt delivery and intensive neonatal support in such situations.

Adult↗

Parvovirus B19 infection reminiscent of myelodysplastic syndrome in three children with chronic hemolytic anemia.

The authors have seen transient pancytopenia with erythroid hypoplasia and striking trilineage myelodysplasia reminiscent of true myelodysplastic syndrome (MDS) in 3 children, 1 with thalassemia intermedia and the other 2 with previously undiagnosed hereditary spherocytosis. In these 3 children transient pancytopenia and myelodysplasia coincided with serological evidence of acute parvovirus-B19 (PV-B19) infection, strongly suggesting their relevance. It is of interest that these 3 cases were encountered within a period of 6 months. This might be an incidental event, but it might also be concluded that acute PV-B19 infection associated transient pancytopenia with morphological appearance of MDS may occur more frequently than reported in the literature. So, PV-B19-associated nonclonal MDS should be considered in the differential diagnosis of true clonal MDS.

Child↗

Parvovirus B19 infection associated with severe aplastic anemia in an immunocompetent patient.

Human parvovirus B19 (PVB19) infection may cause mild pancytopenia characterized by transient and spontaneous recovery in healthy subjects. Severe aplastic anemia associated with PVB19 infection in patients without an underlying disease has been described in a number of reports. Here, a previous healthy, 10-year-old girl with severe aplastic anemia associated with PVB19 infection is described. The patient underwent bone marrow transplantation from her HLA-identical sibling resulting in complete recovery. PVB19 infection should be considered as one of the causes of aplastic anemia in patients without an underlying disease.

Anemia, Aplastic↗

Human parvovirus B19 associated with idiopathic thrombocytopenic purpura.

Although human parvovirus B19 (PVB19) is mainly known to be the causative agent of fifth disease, it is also important in the etiopathogenesis of various hematologic disorders. Recently, its role in the occurrence of chronic anemia and pancytopenia in immunocompromised patients has been revealed. In this study, the authors searched for the presence of PVB19 in a group of patients with idiopathic thrombocytopenic purpura (ITP). The PVB19 DNA was determined by nested PCR, and serologic responses to the virus were evaluated by ELISA. Viral genome was detected by PCR in the sera of 9 of the 19 patients. The IgM and IgG positivities were 57 and 73% in the patients, respectively. These results suggest that PVB19 is associated with ITP nearly in half of the patients and its pathogenetic role in the cases of ITP needs further consideration.

Adolescent↗

Parvovirus B19-associated transient red cell aplasia in children: the role of bone marrow examination in unusual presentations.

Human parvovirus B19 (PV B19) infection in children commonly presents as fifth disease. Transient red cell crisis, the other manifestation of PV B19 infection, is usually reported in children with chronic hemolytic anemia, with a worsening of the anemia. However, this condition may pass unrecognized in children without an underlying hemolytic disorder, since the anemia may be of a short duration and self-limiting. The authors report 3 cases of PV B19-induced transient aplastic in different clinical settings--pancytopenia in one child, during induction phase for acute lymphoblastic leukemia in the second, and fever with joint pains in the third. Treatment for PV B19-induced transient aplastic crisis is essentially supportive. There may be a dilemma in patients on immunosuppressive therapy, since initially it is difficult to distinguish between chronic pure red cell aplasia (a condition where intravenous immunoglobulin therapy is beneficial) and transient aplastic crisis, where supportive red cell transfusions suffice. The patient with leukemia also recovered spontaneously despite being on steroids. In all the 3 patients, the pure red cell aplasia recovered spontaneously without administration of intravenous gammaglobulins.

Bone Marrow Examination↗

Red cell aplasia due to erythrovirus parvovirus B19 infection in a Jehovah's witness with post-transplant EBV-induced aggressive lymphoma: management considerations.

The refusal of Jehovah's Witnesses to accept blood and blood products often poses a clinical dilemma to present day medicine. We present a case of a Jehovah's Witness who had undergone renal transplantation, only to develop an Epstein-Barr virus associated aggressive lymphoma post-transplant. His condition was further complicated by erythrovirus (parvovirus) B 19 infection resulting in red cell aplasia and severe anemia. The management of this difficult clinical situation is discussed together with a review of recommendations for chemotherapy treatment in Jehovah's Witnesses.

Adult↗

Parvovirus B-19 infection during pregnancy.

The development of an acute parvovirus B-19 infection during pregnancy can cause pregnancy complications ranging from early pregnancy loss to nonimmune hydrops. There is no treatment, but preventive measures can be used to decrease perinatal mortality. The diagnosis is made on the basis of clinical suspicion and serology. If the fetus exhibits hydrops in the latter part of pregnancy, the main treatment options include either correcting the associated anemia with intrauterine blood transfusion or birth with extrauterine management. Although the serious problems associated with this virus during pregnancy are uncommon, they can be fatal. In view of this, a pregnant woman who is antibody negative should try to avoid contact with large groups of young children in order to decrease contact with potential vectors.

Adolescent↗

Human parvovirus B19 VP2 empty capsids bind to human villous trophoblast cells in vitro via the globoside receptor.

BACKGROUND: Pregnant women acutely infected with human parvovirus B19 (B19) may transmit the virus to the developing fetus. The mechanism whereby the virus interacts with the placenta is unknown. It is known that globoside receptor is required for successful infection of the target cells, which are the highly undifferentiated, actively dividing colony and burst-form units of the erythroid series. Globoside is present on trophoblast cells which have intimate contact with maternal blood, and may therefore serve as a potential route for B19 transmission into the fetal compartment. OBJECTIVES: The purpose of this study was to determine whether B19 VP2 capsids could bind to villous trophoblast cells in vitro and whether globoside was involved. METHODS: Binding of B19 VP2 empty capsid to first-trimester villous trophoblast cells was assessed by multiple approaches, including ICC using either biotinylated B19 VP2 empty capsid or unlabeled B19 VP2 empty capsid. Quantification of viral binding involved I125-labeled B19 VP2 empty capsid. Competition studies included excess unlabeled empty capsids or pretreatment with globoside-specific IgM antibody. RESULTS: Linear binding of B19 VP2 capsid to purified villous trophoblast cells in vitro was clearly demonstrated (R2= 0.9524). Competition studies revealed specificity of I125-labeled B19 VP2 capsid binding to villous trophoblast cells when pretreatment with either 60-fold excess unlabeled B19 capsid or globoside-specific IgM antibody took place. The results illustrated B19's ability to bind in a specific manner to globoside-containing villous trophoblast cells. CONCLUSION: We speculate that the globoside present on trophoblast cells may play a role in viral binding in vivo, which may facilitate B19 transmission across the maternal-fetal interface.

Binding, Competitive↗

Middle cerebral artery thrombosis in course of parvovirus B19 infection in a young adult: A new risk factor for stroke?

Previous infection, both of bacterial and viral origin, is reported to represent an independent risk factor for ischemic stroke in children and young adults. The authors describe the case of an immunocompetent young woman who developed a middle cerebral artery thrombosis and stroke in course of a recurrence of human parvovirus B19 (PVB19) infection. A previously healthy 25-year-old woman developed right ataxic hemiparesis, 5 days after the onset of a flulike syndrome. Magnetic resonance imaging of the brain revealed acute multiple left frontal-parietal ischemic lesions. Conventional and magnetic resonance angiograms revealed a stenosis in the left middle cerebral artery. Nested polymerase chain reaction detected PVB19-specific DNA sequences in the cerebrospinal fluid and blood, and serology showed high titers of high avidity immunoglobulin G against PVB19. After 10 days, the patient's recovery was nearly complete. One month later, PVB19 disappeared from the serum, whereas it persisted in the peripheral blood mononuclear cells. This case report suggests that PVB19 infection may play a trigger role in the development of ischemic stroke, and that it should be considered in the screening of infectious risk factors for cerebrovascular diseases in young adults.

Adult↗

Viral arthritis: the lessons of parvovirus B19.

Viruses are among the most common and least recognized causes of joint symptoms--including syndromes misclassified as seronegative rheumatoid arthritis. Parvovirus B19 is notably underappreciated in this role. Many cases resolve spontaneously. The challenge is to exclude nonviral causes of joint disease.

Adult↗

Human parvovirus B19 infection in infancy associated with acute and chronic lymphocytic myocarditis and high cytokine levels: report of 3 cases and review.

Human parvovirus B19 infection is occasionally associated with acute lymphocytic myocarditis (ALM). Three infants with B19 virus-associated ALM were followed up clinically, histologically, and immunovirologically. Each infant had B19 virus DNA in the blood or B19 virus-specific IgM antibodies. Two infants with postnatal infection recovered after immunosuppressive therapy. The third infant with possible prenatal infection developed chronic persistent myocarditis associated with persistent B19 virus DNA in the blood. All 3 infants had increased levels of interferon-gamma, tumor necrosis factor-alpha, and interleukins -6 and -8. Four newborns with congenital B19 virus infection and 4 infants and children who had postnatally acquired B19 virus infection without myocarditis all had normal levels of these cytokines. These observations suggest that B19 virus infection in infancy causes ALM in some infants and children.

Acute Disease↗

Prevalence of antibodies to human parvovirus B19 nonstructural protein in persons with various clinical outcomes following B19 infection.

Persistent infections with human parvovirus B19 (B19) associated with debilitating chronic disease have been described, although evidence linking B19 to these more unusual clinical outcomes has been inconclusive. Recent reports have suggested that the development of antibodies to the B19 nonstructural protein (NS1) following B19 infection might be linked to development of severe arthropathy and chronic infection. To confirm these findings, the C-terminal region of the NS1 protein was expressed for use in Western blot assays for detection of anti-NS1 IgG antibodies in human serum. Among 91 persons tested, 0 of 20 not previously infected with B19, 9(36%) of 25 with past B19 infection, and 5 (12.5%) of 40 with recent B19 infection, had detectable anti-NS1 antibodies. Of 6 persons with chronic B19 infection, 2 had detectable antibodies to NS1. The presence of anti-NS1 antibodies did not appear to correlate with unusual clinical outcomes or chronic B19 infection.

Animals↗

T helper cell-mediated in vitro responses of recently and remotely infected subjects to a candidate recombinant vaccine for human parvovirus b19.

T cell proliferation to human parvovirus B19 antigen was measured in 6 patients with recent B19 infection (1 with pneumonia and pleuritis), 1 patient with symptoms persisting >180 days after onset, 18 nonsymptomatic subjects with remote B19 immunity, and 12 B19-seronegative control subjects. Recombinantly expressed virus-like particles (VP1/2 capsids), a candidate B19 vaccine, were used as antigen. Virus-specific T helper cell proliferation was detectable in all the recently infected patients and in most (17/18) of the remotely infected subjects but not in the seronegative control subjects. The B19-specific T cell responses, in general, were most vigorous among the recently infected patients. However, such strong B19-specific proliferation was not confined within the acute phase, as 28% (5/18) of the remotely infected healthy individuals had B19-specific reactivity persisting at acute-phase levels, apparently for years or decades. These data indicate that B cells recognizing the VP1/2 capsids receive class II-restricted help from CD4(+) T lymphocytes.

Acute-Phase Reaction↗