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PubMed2000

Parvovirus B19 transmitted by bone marrow.

We describe a case of symptomatic parvovirus B19 infection transmitted by bone marrow (BM). The infection caused prolonged anaemia, thrombocytopenia, arthralgia and erythema infectiosum in a 16-year-old girl with acute myeloid leukaemia receiving a BM transplant (BMT). The BM donor was a 19-year-old asymptomatic brother who had parvovirus B19 viraemia at the time of BM harvest. Sequencing of the VP2 gene from the patient and the donor showed a perfect match of DNA sequences, confirming the mode of transmission. Parvovirus B19 represents a potential complicating factor in patients undergoing BMT, but screening by polymerase chain reaction (PCR) of donor BM may reduce the risk of infection.

Adolescent↗

PubMed2000

Prevalence of human parvovirus B19 and TT virus in a group of young haemophiliacs in South Africa.

A well recognized hazard of transfusion with blood or blood products is the acquisition of a viral infection. Parvovirus B19 and transfusion transmitted virus (TTV) are two of several non-enveloped viruses that may on rare occasions be present in coagulation factor concentrates. The prevalence of these viruses in the South African Haemophilia population has not previously been studied. Thirty-nine Haemophiliac children were investigated for evidence of parvovirus and TTV infection. 26 boys with Haemophilia A had been treated with cryoprecipitate or intermediate purity factor VIII, and 13 boys with Haemophilia B had received prothrombin complex concentrates. All the plasma products were prepared from South African donors and were virally inactivated by heat or solvent/detergent since 1992. A control group of 32 children who had not been transfused were also studied. IgG antibodies to B19 were present in 29 of the 39 patients (74%), 18/26 (69%) with Haemophilia A and 12 of the 13 (85%) with Haemophilia B. None of the patients was IgM antibody positive but two children were PCR positive for B19 DNA. Of the control children, 47% had IgG antibodies to B19, but none were IgM antibody or B19 DNA positive. TTV viral DNA was found in 10.2% of patients and in 9% of the control group. The results indicate that our locally produced plasma products are not a significant source of TTV transmitted infection but may contribute to infection by B19 parvovirus.

Adolescent↗

PubMed2001

Parvovirus B19 DNA in plasma pools and plasma derivatives.

BACKGROUND AND OBJECTIVES: Human parvovirus B19 (B19) has been transmitted by various plasma-derived medicinal products. The aim of this study was to determine the frequency and the level of B19 DNA contamination in plasma pools destined for fractionation and in a broad range of plasma derivatives. In addition, removal of B19 DNA by the manufacturing process was investigated in cases where corresponding samples from plasma pool and product were available. MATERIALS AND METHODS: Plasma pool samples and blood products were tested for B19 DNA by nested polymerase chain reaction (PCR), and the viral DNA content was determined by TaqMan quantitative PCR. RESULTS: Two-hundred and twenty two of 372 plasma pools for fractionation contained B19 DNA at concentrations of 10(2)-10(8) genome equivalents/ml (geq/ml). While approximately 65% of the DNA-positive plasma pools were only moderately contaminated (< 10(5) geq/ml), 35% contained > 10(6) geq/ml. High frequencies of contamination were detected in Factor VIII (79 of 91), prothrombin complex concentrates (38 of 43) and Factor IX (41 of 62), where the concentration of B19 DNA ranged between 102 and 107 geq/ml. A lower level of B19 DNA contamination was found in antithrombin III (five of 26 samples), in anti-D immunoglobulins (three of 37 samples) and in albumin (four of 51 samples), with levels ranging between 10(2) and 10(3) geq/ml. Furthermore, investigation of plasma pools for solvent/detergent plasma (S/D plasma), from two manufacturers, revealed B19 DNA in 15 of 66 batches at concentrations of 10(2)-10(8) geq/ml. Similar concentrations were detected in the corresponding final S/D plasma products. Anti-B19 immunoglobulin G (IgG) was found in plasma pools and S/D plasma at concentrations of approximately 40 IU/ml. CONCLUSION: Although positive PCR results do not necessarily reflect infectivity, these data show that B19 is a common contaminant in plasma pools and in plasma-derived medicinal products. Considering the resistance of animal parvoviruses to inactivation by heat and chemical agents, and the absence of specific information for B19, the risk of B19 transmission by plasma products should be considered. Physicians should be aware of this problem when treating patients of B19-related risk groups. The plasma fractionation industry should continue their efforts to avoid B19 contamination of plasma derivatives and develop methods which are effective in removing/inactivating parvovirus B19.

Blood↗

PubMed2000

Apoptosis in feline panleukopenia and canine parvovirus enteritis.

Tissue samples of cats and dogs with panleukopenia and parvovirus enteritis, respectively, were examined for the presence of viral antigen-positive cells and apoptotic cells by immunohistochemistry and by TUNEL assay (Terminal Transferase-Mediated dUTP Nick End Labelling). Compared to control animals, infected cats and dogs generally had more TUNEL-positive cells. Cell types positive for parvovirus antigen, for example digestive tract epithelial and mesenchymal cells, and lymphocytes and macrophages in lymphoid tissues were also positive for TUNEL signals. Occasionally, TUNEL signal and viral antigen were present in the same tissue areas, suggesting a direct viral trigger of apoptosis. More frequently, however, there was no complete overlap of antigen and TUNEL-positive areas. The results of this study indicate that apoptotic cell death contributes significantly to the widespread tissue damage of parvovirus infection in cats and dogs.

Animals↗

PubMed2002

Removal of neutralized model parvoviruses and enteroviruses in human IgG solutions by nanofiltration.

BACKGROUND: Nanofiltration has proven to be an effective technology for virus removal. The small nonenveloped viruses, such as HAV and parvovirus B19 (B19), are evidently more difficult to remove than the larger enveloped viruses. However, since antibodies against these viruses, HAV and B19, are constantly present in large plasma pools, it is conceivable that antibody binding will lead to an increase in size of the viruses due to a corona of antibodies on the virus surfaces. These enlarged viruses should be easier to remove by nanofiltration than naked viruses. In this study, antibody-coated viruses and free virions were subjected to nanofiltration to determine whether the antibody-mediated increase in size contributes to the elimination of viruses which are otherwise too small to be retained by filters of a given nominal pore diameter. STUDY DESIGN AND METHODS: One-percent IgG or 1-percent albumin solutions were spiked with bovine enterovirus (BEV) and filtered through 50-nm filters. This virus cross-reacts with antibodies in pooled human IgG solutions but persists as free virions in albumin solutions. Similarly, two parvoviruses, bovine parvovirus (BPV) and minute virus of mice (MVM), in 1-percent IgG solutions were filtered through 20-nm filters. BPV cross-reacts with antibodies in human IgG, whereas MVM remains unaffected. Virus removal was assessed by in vitro infectivity assays. Two methods were developed to displace the antibodies from BEV and BPV and thereby render them accessible for titration. RESULTS: Antibody-coated BEV was eliminated to below detection limit by the 50-nm filters (logarithmic reduction factors [LRF] > or = 5), while free BEV virions passed without restraint through the same filters. Similarly, antibody-coated BPV was completely retained by the 20-nm filters (LRF > or = 5). MVM virions were strongly retained by the 20-nm filters; however, the virus was still detectable in the filtrate. CONCLUSION: These results indicate that viruses with bound antibodies are efficiently eliminated by nanofiltration by use of filters having nominal pore sizes larger than the diameter of the respective free virions.

Enterovirus↗

PubMed2001

Fatal parvovirus myocarditis in a 5-year-old girl.

Infection with parvovirus B19 is common in children and typically causes mild illness. We report here the case of a 5-year-old girl who died suddenly, 2 weeks after the clinical diagnosis of a parvoviral infection (erythema infectiosum). Microscopic examination of the heart showed severe myocarditis with extensive T-cell and macrophage infiltration. Cultures, serology, and molecular analyses of serum for enteroviridae, adenovirus, influenza, varicella zoster, cytomegalovirus, and herpes simplex viruses were negative. Quantitative polymerase chain reaction (PCR) analysis for parvovirus B19 in peripheral blood, however, showed active infection (91,000 genomes/mL serum; 2.4 genomes/mononuclear cell). Despite the presence of myocarditis, immunostaining for parvoviral surface antigens was negative in the heart. Quantitative PCR analysis of paraffin sections showed that myocardial parvoviral content was significantly less than that of the normal appearing kidney and within the range predicted simply by tissue blood content. Thus, parvovirus B19 infection can be complicated by fatal myocarditis. Because the virus does not appear to have infected the heart, per se, we speculate that myocarditis arose from immunological cross-reaction to epitopes shared between the virus and the myocardium. HUM PATHOL 32:342-345.

Antigens, Viral↗

PubMed1999

Hydranencephaly and cerebellar hypoplasia in two kittens attributed to intrauterine parvovirus infection.

Six weeks after vaccination with modified live feline parvovirus vaccine, a cat gave birth to five kittens, three of which died soon afterwards. The remaining two kittens (A and B) survived, but at 8 weeks of age were unable to walk and showed abnormal behaviour, with lack of menace and oculovestibular responses, and severe dysmetria. These signs suggested multifocal disease associated with the cerebrum and cerebellum. Magnetic resonance imaging demonstrated severe bilateral (kitten A) or unilateral (kitten B) hydrocephalus or hydranencephaly, combined with cerebellar agenesis (kitten A) or severe hypoplasia (kitten B). Hydranencephaly was confirmed histopathologically in both kittens. Parvovirus was isolated from the kidney of one kitten. Parvoviral DNA was amplified by the polymerase chain reaction (PCR) from paraffin wax-embedded brain of both kittens. The severe malformations observed in these kittens presumably resulted from an in-utero parvovirus infection, possibly due to vaccination, that occurred late in the first, or early in the second, trimester of pregnancy.

Animals↗

PubMed2002

[Parvovirus B19 myocarditis in a young man with previous non-bacterial meningitis].

HISTORY AND CLINICAL FINDINGS: A 22-year old man presented with fatigue, dyspnea NYHA III and presyncopes that had persisted since a non-bacterial meningitis 3 months before. INVESTIGATIONS: Transthoracic echocardiography revealed a dilated left ventricle with an ejection fraction (EF) reduced to 35-40 % due to global hypokinesia. No pericardial effusion was seen; ECG and lung function test were normal. Serological, immunological and microbiological tests as well as nested PCR analysis of blood leucocytes for detection of cardiotropic pathogens were inconclusive. In endomyocardial biopsies retrieved from the left ventricular posterolateral wall, a chronic macro-phage-rich myocarditis was shown by histopathology and, in addition, Parvovirus B19 was identified as specific pathogen by use of nested PCR analysis. TREATMENT AND COURSE: At physical rest and with ACE inhibitor therapy (2.5 mg ramipril/day), heart failure decreased steadily. Follow-up echocardiography 1 month later revealed a left ventricle that was only slightly dilated with an EF of 50 %. 3 months later, the patient was markedly more load-bearing; the EF amounted to 55-60 %. CONCLUSIONS: Parvovirus B19 should be regarded as potential pathogen in case of suspected myocarditis in adulthood. Whether the previous non-bacterial meningitis was also attributable to this specific pathogen, remains open. Of note, however, the present case report by demonstrating a localized myocardial Parvovirus B19 infection without detectable systemic infection underscores the importance of molecular tests for diagnostic accuracy in manifest organ failure.

Adult↗

PubMed2004

The structure of human parvovirus B19.

Human parvovirus B19 is the only parvovirus known to be a human pathogen. The structure of recombinant B19-like particles has been determined to approximately 3.5-A resolution by x-ray crystallography and, to our knowledge, represents the first near-atomic structure of an Erythrovirus. The polypeptide fold of the major capsid protein VP2 is a "jelly roll" with a beta-barrel motif similar to that found in many icosahedral viruses. The large loops connecting the strands of the beta-barrel form surface features that differentiate B19 from other parvoviruses. Although B19 VP2 has only 26% sequence identity to VP3 of adeno-associated virus, 72% of the C(alpha) atoms can be aligned structurally with a rms deviation of 1.8 A. Both viruses require an integrin as a coreceptor, and conserved surface features suggest a common receptor-binding region.

Capsid Proteins↗

PubMed1998

Parvovirus B19 infection associated with myocarditis following adult cardiac transplantation.

A 56-year-old woman underwent an uneventful cardiac transplantation due to dilated cardiomyopathy. One week later the patient developed clinical and histological signs of myocarditis. We report for the first time a case of myocarditis in an adult heart transplant recipient, possibly induced by parvovirus B19, as evidenced by the finding of specific IgM in serum and specific DNA in the myocardial cells. Furthermore, this is the first time parvovirus B19 DNA has been observed in the myocardium of an adult. In conclusion, parvovirus B19 should be recognized as a potential pathogen causing myocarditis in heart transplant recipients. In order to establish a definite and rapid diagnosis, a search for specific IgM should be supplemented with PCR investigations of serum and myocardial biopsies when available.

Antibodies, Viral↗

PubMed1996

Seroprevalence of human parvovirus B19 in children of a desert region.

A seroepidemiological study was conducted in Kuwait to evaluate the pattern of acquisition of human parvovirus B19 by children in Kuwait and to compare it with patterns described in other regions in different climatic zones. A total of 218 serum samples from children less than 16 years of age were tested for the presence of anti-B19 IgG by enzyme immunoassay. The overall seroprevalence was 17.4%. Infants in Kuwait had low levels of maternally-acquired anti-B19 IgG (8.6%). The age of peak exposure to parvovirus B19 was 10-15 years compared with less than 10 years in England and Wales and more than 20 years in Singapore. The results of this study indicate an influence of geographic differences on transmission of parvovirus B19.

Antibodies, Viral↗

PubMed2005

Expression of capsid proteins and non-structural proteins of waterfowl parvoviruses in Escherichia coli and their use in serological assays.

While there are a number of methods available for detection of antibodies against waterfowl parvoviruses, none is able to differentiate responses against the capsid and non-structural proteins. To enable this, the capsid and non-structural proteins of goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV) were expressed in Escherichia coli. These proteins were purified and used as antigens in western blotting assays of antibodies against GPV and MDPV. The results showed that 94.7% of the goose and 90.0% of the duck sera collected from the field contained antibodies against GPV or MDPV. Moreover, these sera could be classified into distinct groups based on differences in patterns of western blot reactivity. These different patterns might indicate different stages in infection. Western blotting assays of sera collected from experimentally infected ducks showed that antibodies against the non-structural protein appeared first after infection, followed by antibodies against the capsid protein. It was concluded that the recombinant capsid and non-structural proteins might serve as useful antigens for assays for antibodies against GPV and MDPV. Moreover, because these assays could discriminate between antibodies against the non-structural protein and those against the capsid protein, they may be useful in differentiating vaccinated from infected birds when recombinant capsid protein is used as the vaccine.

Animals↗

PubMed1998

IgG response to the immunoreactive region of parvovirus B19 nonstructural protein by immunoblot assay with a recombinant antigen.

A total of 190 serum samples from patients with different clinical manifestations in whom B19 parvovirus infection was suspected and 108 control serum samples were analyzed for the IgG response against parvovirus B19 nonstructural protein. An immunoblot assay was developed using the immunoreactive region of the B19 nonstructural protein as recombinant antigen. Virologic (B19 DNA) and serologic (IgM and IgG anti-VP) markers of B19 infection were also investigated. There was an even distribution of IgG nonstructural antibodies in serum samples of parvovirus B19-infected patients and controls (27.7% and 21.7%, respectively) that were mainly associated with patterns of past B19 infection.

Antibody Formation↗

PubMed2004

Simian parvovirus infection: a potential zoonosis.

INTRODUCTION: Simian parvovirus (SPV) causes severe anemia in immunocompromised macaques. The closely related erythrovirus, parvovirus B19, causes anemia in susceptible humans and can be grown in human bone marrow mononuclear cells in vitro. We hypothesized that SPV may infect humans and replicate in human bone marrow mononuclear cells. METHODS: Serum samples from handlers of an SPV-seropositive macaque colony were tested by Western blot for evidence of antibodies to SPV. SPV capsid protein was expressed in insect cells, and SPV was cultured in human and macaque bone marrow mononuclear cells. RESULTS: Fifty-one percent of exposed handlers (n=65) were found to be SPV seropositive, compared with 35% of nonexposed individuals (n=124). In 17% of exposed handlers, compared with 6% of nonexposed individuals, antibodies were directed to SPV but not to B19. SPV capsid proteins, like those of B19, self-assembled to form parvovirus-like particles, and these capsids, like B19 capsids, bound to globoside, suggesting that globoside is also the receptor for SPV. We demonstrated that SPV could replicate in vitro in both human and macaque bone marrow mononuclear cells and that it was cytotoxic to erythroid progenitor cells. CONCLUSIONS: Our data suggest that SPV may infect human bone marrow mononuclear cells in vitro and in vivo and should be considered a potential zoonosis.

Animals↗

PubMed2000

Highly efficient transduction and expression of cytokine genes in human tumor cells by means of autonomous parvovirus vectors; generation of antitumor responses in recipient mice.

The possible use of recombinant autonomous parvoviruses as vectors to efficiently express therapeutic cytokines in human tumor cells was evaluated in vitro and in vivo. The parvovirus H1 was used to generate recombinant viruses (rH1) that carried transgenes encoding either human interleukin 2 (IL-2) or monocyte chemotactic protein 1 (MCP-1), in replacement of part of the capsid genes. Such rH11 viruses have been shown to retain in vitro the intrinsic oncotropic properties of the parental virus. On infection with the recombinant viruses at an input multiplicity of 1 replication unit (RU) per cell, HeLa cultures were induced to release 4-10 microg of cytokine per 10(6) cells over a period of 5 days. The expression of the rH1-transduced human cytokine/chemokine could also be detected in tumor material recovered from nude mice that had been subcutaneously engrafted with in vitro-infected HeLa cells. The formation of tumors from HeLa xenografts was reduced by 90% compared with wild-type or mock-infected cells as a result of cells preinfected with IL2-expressing virus at an input multiplicity as low as 1 RU per cell. Tumors arising from HeLa cells infected with transgene-free or MCP1-expressing vectors or with wild-type H1 virus were not rejected at this virus dose. Tumors infected with rH1/IL-2 virus displayed markers indicative of their infiltration with NK cells in which the cytocidal program was activated, whereas little NK activity was detected in wild-type virus or mock-infected tumors. Altogether, these data show that the IL-2 expressing H1 vector was a more potent antineoplastic agent than the parental virus, and point to the possible application of recombinant autonomous parvoviruses toward therapy of some human tumors.

Animals↗

PubMed1993

Candidate recombinant vaccine for human B19 parvovirus.

Recombinant baculoviruses were used to produce human B19 parvovirus empty capsids composed of only VP2 and VP2 capsids containing 4%, 25%, 35%, or 41% VP1 protein. Immunogenicity of the purified capsids, formulated with or without adjuvant, was evaluated in mice, guinea pigs, and rabbits. Sera were analyzed for total anti-B19 parvovirus antibodies, antibodies specific to the region unique to the VP1 capsid protein, and virus neutralizing antibodies. A relationship was observed between the development of antibodies specific to sequences unique to the VP1 protein and virus neutralization. The polypeptide composition of the empty capsid immunogens appeared to be important for elicitation of potent virus neutralizing activity. VP2 capsid immunogens devoid of VP1 protein, or consisting of only 4% VP1, the composition of naturally occurring virions, were generally poor at eliciting high levels of virus neutralizing activity. Capsids consisting of > or = 25% VP1 protein efficiently and consistently provoked vigorous B19 virus neutralizing responses. Recombinant empty capsids enriched for the VP1 protein should serve as the basis for a human B19 parvovirus vaccine.

Aluminum Hydroxide↗

PubMed1993

Generation of neutralizing anti-B19 parvovirus human monoclonal antibodies from patients infected with human immunodeficiency virus.

The prevalence of IgG antibodies to human B19 parvovirus (anti-B19) is elevated in individuals infected with human immunodeficiency virus (HIV), especially during the later stages of HIV infection. In subjects with high titers of IgG anti-B19, 86% (19 of 22) had circulating B cells producing anti-B19. Immortalization of these cells with Epstein-Barr virus and generation of heterohybridomas by fusion with a mouse X human heteromyeloma resulted in the production of two cell lines producing IgG1 kappa monoclonal antibodies (MAbs). Both of these MAbs were specific for conformational epitopes on the VP2 capsid protein of B19 parvovirus and both were capable of neutralizing 50% of the viral infectivity in a human erythroid colony-forming unit assay at < or = 1 micrograms of MAb/mL. These human MAbs are potentially useful in the treatment of acute B19 parvovirus infection.

Antibodies, Monoclonal↗

PubMed1995

Multiple glycosphingolipids determine the tissue tropism of parvovirus B19.

Infection with human parvovirus B19, the etiologic agent of fifth disease, is associated with numerous hematologic and nonhematologic complications. Recently, the receptor for parvovirus B19 was reported to be globoside (Gb4), a neutral glycosphingolipid (GSL) of red cell membranes. To ascertain if tissue Gb4 expression correlates with B19-associated disease, neutral GSLs from 16 human tissues were isolated and analyzed using high-performance thin-layer chromatography and immunostaining with anti-Gb4 monoclonal antibodies or B19 empty capsids. Gb4 was identified as a major neutral GSL in 11 tissues, especially in those of mesodermal origin. In addition to recognizing Gb4, B19 capsid bound to several tissue-specific GSLs, including two complex globo series GSLs (SSEA-3, SSEA-4) and paragloboside (neolactotetraglycosylceramide), as was demonstrated in red cell, granulocyte, kidney, liver, and bowel tissue. There was good correlation between tissue-neutral GSL expression, B19 capsid binding, and the tissue tropism observed clinically in B19 parvovirus-associated disease.

Blood Cells↗