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PubMed2005

Chronic parvovirus infection and G6PD deficiency masquerading as Diamond-Blackfan anemia.

Bone marrow aplasia due to parvovirus B19 infections is usually mild and self-limited. In patients with hereditary or acquired hemolytic anemias, B19 infection can cause a severe and life-threatening anemia due to the shortened half-life of red cells, but here, too, the transient nature of the infection soon remits the symptoms. Chronic infections with parvovirus are more characteristically associated with immunodeficiency states. We report here a case of B19-induced anemia in a patient with G6PD deficiency and hypogammaglobulinemia which was mislabeled as Diamond-Blackfan anemia until the use of modern laboratory techniques allowed a correct diagnosis to be made.

Adult↗
PubMed1990

Rheumatologic manifestations of human parvovirus B19 infection in adults. Initial two-year clinical experience.

During 1987 and 1988, we identified 9 adults at the Medical and Rheumatology Services of the University of Iowa Hospitals and Clinics who had a clinical diagnosis of fifth disease; 8 of the 9 had symptoms of joint involvement. Another 12 adults with serologic positivity for anti-parvovirus B19 IgM antibody presented with polyarthralgia/polyarthritis. Patients were usually found to be seronegative for rheumatoid factor, and none developed nodules or erosive disease. Many patients with chronic disease met criteria for a diagnosis of rheumatoid arthritis. A diagnosis of parvovirus B19 infection should be considered during the initial visit of patients with polyarthralgia/polyarthritis.

Adult↗
PubMed2002

Enhanced kinetic extraction of parvovirus B19 structural proteins.

Recombinant structural proteins (VP1 and VP2) of the human parvovirus B19 have been expressed simultaneously using the baculovirus expression system to form virus-like particles (VLPs) that have potential use as vaccines. In this study, we report optimization of extraction conditions to recover these VLPs from cell paste. Under hypotonic conditions with neutral pH these VLPs were poorly extracted (up to 3% extraction). Addition of reducing agents, detergents, salts, and sonication did not improve the extractability. While screening for conditions to improve the extractability of the VLPs, we discovered that a combination of higher pH and elevated processing temperature significantly increased the extraction. Whereas increasing pH alone increased extractability from 3% to 6% (pH increased from 8.0 to 9.5), the effect of elevated temperature was much more substantial. At 50 degrees C, we observed the extraction to be more than fivefold higher than that at room temperature (up to 25% extracted at pH 9.0). The kinetics of extraction at elevated temperatures showed a rapid initial rate of extraction (on the order of minutes) followed by a plateau. In addition, we compared the extraction of VP1 expressed alone. VP1 expressed alone is incapable of forming VLPs. We observed that non-VLP VP1 was easily extractable (up to 60% extracted) under conditions in which the VP1 + VP2 VLPs were not extractable. From these studies we conclude that parvovirus B19 structural proteins expressed to form VLPs have a hindered extractability as compared with non-VLP protein. This hindrance to extraction can be significantly reduced by processing at elevated temperatures and an increased pH, possibly due to the enhanced rates of solubilization and diffusion.

Animals↗
PubMed1985

Detection of human parvovirus using a molecularly cloned probe.

Half of the genomic DNA of the human parvovirus (B19) was cloned in the plasmid pBR322. The cloned DNA was used as a molecular probe for the detection of parvovirus in serum by means of a dot hybridization test. In an assay of 26 samples, the dot hybridization test was found to be of comparable sensitivity and to be as rapid as radioimmunoassay for viral antigen detection; it is potentially useful as a diagnostic test.

Cloning, Molecular↗
PubMed1987

Detection of parvovirus B19 DNA, antigen, and particles in the human fetus.

Human parvovirus B19 commonly infects children, causing erythema infectiosum (fifth disease). However, there is a significant adult population which has not been exposed to the virus and, consequently, does not have protective antibody. Recent reports have associated B19 infection during pregnancy with fetal death, although normal outcome of pregnancy is more common. To characterise further the role of B19 infection in fetal deaths, a series of laboratory investigations has been undertaken on tissues obtained at autopsy. These have demonstrated the presence of virion-sized DNA by Southern blotting, viral antigen by radioimmunoassay, and viral particles by electron microscopy, all from tissues of hydrops fetalis. These data confirm that the human parvovirus B19 can cross the placenta and replicate in fetal tissues.

Antigens, Viral↗
PubMed1990

Electron microscopic detection of human parvovirus (B19) in a patient with HIV infection.

We report the findings on an HIV-positive patient found to be infected with human parvovirus B19. In a comparative study of sequential sera, direct electron microscopy (EM) had the same sensitivity as DNA hydridization for the detection of the virus. EM did not require specific reagents and also yielded information on the state of the virus; i.e., whether random or complexed, and the type of antibody involved. The presence of parvovirus complicated the judgement as to drug treatment. This case also highlighted the importance of diagnosing the presence of opportunistic viral infections that have no, or low-grade, pathogenic activity.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Prevalence of human parvovirus (B19) and rubella virus infections in urban and remote rural areas in northern Brazil.

Sera from inhabitants of Belém, Pará (542 sera), Brazil and of members of 3 Brazilian tribes--Tiriyo/Alto Paru (near Surinam) (212 sera), Xicrin (128 sera), and Mekranoiti (121 sera)--of different age and sex groups were tested for the presence of specific antibody against human parvovirus (B19) (RIA) and rubella virus (latex agglutination test). Parvovirus (B19) IgG was found in 42.6% of the population sample from Belém but in only 4.7 to 10.7% of the members of the tribes. Rubella virus antibody was found in 72.7% of the sera from Belém but approaching a prevalence of 85-90% in age groups above 20 years. In the tribes rubella virus antibody was detected in 36.9 to 72.6% of all sera. There were remarkable sex differences of antibody prevalence in several age groups of the population from Belém and of the tribal populations. About a quarter of the skin rashes in Belém that were not attributable to infections with rubella, measles, or arboviruses were caused by recent B19 infections.

Adolescent↗
PubMed1989

Embryonic malformations in a case of intrauterine parvovirus B19 infection.

An incomplete embryo of 9 weeks development from a woman infected by human parvovirus B19 during early pregnancy was histologically examined. B19-DNA was detected in both embryonic and placental tissue by dot-blot hybridization. Focal vascular endothelial damage was found throughout the entire embryo and placenta together with mononuclear infiltrations around the vessels. In the placenta these mononuclear cells belonged for the greater part to the cytotoxic and/or suppressor T-cell group. One eye showed lens abnormalities and retinal folds. The other eye was microphthalmic and aphakic and showed dysplasia of the sclera, anterior segment, and retina. The skeletal muscle cells displayed a general eosinophilic degeneration. Focally, similar changes were found in heart muscle and smooth muscle tissue. In several tissues pathologic effects at a cellular level were noted, as intranuclear vacuole-like inclusions and nuclear ballooning. On the basis of this study it is concluded that human parvovirus B19 may affect several fetal tissues and may even have teratological effects on a developing human embryo.

Abortion, Legal↗
PubMed1993

The canine parvovirus empty capsid structure.

The structure of empty canine parvovirus capsids shows that residues 37 to the carboxy-terminal residue 584 (VP2 numbering) are ordered in each of the 60 subunits. The central structural motif of each subunit is the eight-stranded antiparallel beta-barrel that has been found in many other virus structures. Five beta-hairpin turns form a beta-cylindrical structure at each icosahedral 5-fold axis. The N-terminal glycine-rich sequence can be accommodated within this cylinder without excessive steric hindrance, consistent with the electron density distribution. By far the largest conformational differences between the full and empty virus were found in the region where some ordered DNA has been observed to bind in canine parvovirus full particles. Extensive interactions among 3-fold related subunits indicate that a trimeric subunit might be a viral assembly intermediate.

Amino Acid Sequence↗
PubMed1993

Mutational analysis of conserved tyrosines in the NS-1 protein of the parvovirus minute virus of mice.

The NS-1 gene of the parvovirus minute virus of mice encodes a multifunctional protein essential for viral DNA replication and gene expression. In addition to possessing DNA helicase and ATPase activities, NS-1 forms a covalent linkage with the 5' ends of viral DNA and is a strong candidate for the site-specific nicking-closing enzyme postulated to be involved in the resolution of concatemers and terminal hairpin structures that arise during parvoviral DNA replication. Since the covalent linkage between NS-1 and the 5' terminus of MVM DNA resists alkali and mild acid treatment, a tyrosine phosphodiester is likely to be involved. To map domains responsible for this activity, mutations converting tyrosine to phenylalanine were introduced into the NS-1 gene using oligonucleotide-directed mutagenesis and their effect on the DNA replication and transcriptional activation functions of NS-1 was examined in transient in vivo transfection assays. Replacement of Tyr-188, Tyr-197, Tyr-210, Tyr-310, Tyr-422, or Tyr-550 with phenylalanine greatly reduced the ability of NS-1 to complement the replication of the target genome ins 20B in COS-7 cells. However, a Ser-545 to Thr-545 substitution in the Phe-550 mutant restored DNA replication activity. Replacement of 5 other tyrosines in NS-1 with phenylalanine either enhanced (Phe-6), had a moderate inhibitory effect (Phe-209) or had no effect (Phe-47, Phe 227 and Phe-543) on its DNA replication activity. Two of the 11 phenylalanine substitution mutations, Phe-188 and Phe-197, also greatly reduced the ability of NS-1 to transactivate the p38 promoter and displayed a dominant negative phenotype with respect to transactivation. Since the remaining tyrosines in MVM NS-1, Tyr-152, Tyr-252, Tyr-374, and Tyr-595, are not conserved among the NS-1 proteins encoded by porcine and feline parvoviruses, they are presumed to be nonessential for the normal functioning of NS-1. The results point to a role for either Tyr-188, Tyr-197, Tyr-210, Tyr-310, or Tyr-422 in forming a covalent linkage with viral DNA and further suggest a regulatory role for several tyrosines in other DNA replication and transcriptional activation functions of NS-1.

Amino Acid Sequence↗
PubMed1993

Construction of an infectious DNA clone of the Y1 strain of canine parvovirus and characterization of the virus derived from the clone.

We have cloned genome fragments of canine parvovirus strain Y1 from replicative-form DNA and double-stranded DNA synthesized from virion DNA in vitro, and constructed a recombinant plasmid containing a full-length Y1 genome (pCPVY 1). When this recombinant plasmid was transfected into cell cultures, an infectious virus could be recovered. To characterize this pCPVY 1-derived virus, its biological properties were compared with those of the parental strain. No difference was observed between them in antigen expression, viral DNA replication, hemagglutination ability, and virus multiplication, indicating that the virus derived from the infectious plasmid inherited the biological properties of the authentic Y1 strain. Therefore, this recombinant plasmid appears to be useful for reverse genetics of canine parvovirus.

Animals↗
PubMed1980

Astrovirus-like, coronavirus-like, and parvovirus-like particles detected in the diarrheal stools of beagle pups.

Astrovirus-like, coronavirus-like, and parvovirus-like particles were detected through electron microscopic (EM) examination of loose and diarrheal stools from a litter of beagle pups. Banding patterns obtained from equilibrium centrifugations in CsCl supported the EM identification. Densities associated with the identified particles were: 1.34 g/ml for astrovirus, 1.39 g/ml for "full" parvovirus and 1.24-1.26 g/ml for "typical" coronavirus. Convalescent sera from the pups aggregated these three particle types as observed by immunoelectron microscopy (IEM). Only coronavirus-like particles were later detected in formed stools from these same pups. Coronavirus and parvo-like viruses are recognized agents of canine viral enteritis, however, astrovirus has not been previously reported in dogs.

Animals↗
PubMed1992

A monoclonal antibody which recognizes cell surface antigen and inhibits porcine parvovirus replication.

Monoclonal antibody technologies were applied to the study of early events in porcine parvovirus (PPV) infections in vitro. Balb/c mice were immunized with whole swine testicle cells and hybridomas were produced following fusion with myeloma cells. Resultant clones were screened firstly in an ELISA system, to detect monoclonal antibody recognition of swine testicle cells, and secondly, in a fluorescent antibody test to detect monoclonal antibody which inhibited production of PPV antigen. One clone, 1H11, which satisfied these screening requirements, recognized proteins present in cell lines both permissive and non-permissive for porcine parvovirus replication and inhibited the production of virus progeny of several PPV isolates. A linear staining pattern of cross-linked plasma membranes, indicative of monoclonal antibody binding at the cell membrane, was demonstrated by indirect immunofluorescence assays. In immunoblotting experiments, 1H11 recognized a polypeptide of approximately 40 kDa in size, present in both permissive and non-permissive cell lines.

Animals↗
PubMed1980

The association of calf serum with the contamination of BHK21 clone 13 suspension cells by a parvovirus serologically related to the minute virus of mice (MVM).

An investigation of persistent cell deaths of BHK21 suspension cells during subculturing resulted in the isolation of a viral agent. The agent was isolated from samples of dead cells, cell growth media and 2 batches of calf serum. It was established that the agent was associated with the use of certain batches of calf serum. The isolated virus was found to replicate effectively only in rapidly growing BHK cell cultures. In monolayers it caused the formation of large intranuclear inclusion bodies. The isolate was a strong haemagglutinin; it was stable to ether, chloroform, pH 3 and heating at 56 degrees C. It was shown to be a DNA-virus and by electron-microscopy it was evident as unenveloped spherical, small particles (21 nm in diameter). In sucrose density gradients the virus sedimented as 2 peaks at approximately 114S and 85-92S, and by caesium chloride equilibrium centrifugation, two main peaks at densities of 1.39 and 1.31 g/ml were evident with a minor peak at 1.35. The capsid of the complete virion consisted of 3 polypeptides. The agent has therefore been provisionally designated a member of the Parvoviridae family, genus parvovirus. Serologically it was found to be related to MVM. The isolated parvovirus was inactivated by 0.05 per cent peracetic acid and 0.05 per cent acetylethyleneimine.

Animals↗
PubMed1980

Transplacental infection and embryonic death following maternal exposure to porcine parvovirus near the time of conception.

Each of 20 gilts (principals) from a commercial swine herd free of antibody for porcine parvovirus (PPV) was exposed intranasally and orally to PPV at the onset of gestation. The gilts were killed and necropsied 22 +/- 1 days later to determine the effect of the virus on their embryos. An equal number of gilts (controls of the same status, from the same herd, and bred to the same boars, were treated similarly except for exposure to PPV. The following data were obtained at necropsy and from subsequent laboratory tests. Principals had 223 corpora lutea (8 to 16/gilt) and 203 embryos (6 to 16/litter). Porcine parvovirus-infected embryos (1 to 9/litter) were detected in 12 (60 per cent) of the 20 litters. Of the 203 embryos of principals, 169 were alive and 34 were dead and in various stages of decomposition and resorption. Both virus and viral antigen were detected in 7 of the live embryos (1, 1, 2, and 3 in 4 litters) and in 32 of the dead embryos. With 1 exception, infected live embryos were next to infected dead littermates in the uterus suggesting the beginning of intrauterine spread of the virus. Controls had 234 corpora lutea (9 to 15/gilt) and 217 embryos (9 to 14/litter). None of their embryos were infected with PPV and all but 3 were alive.

Animals↗
PubMed1991

[Hydrops fetalis caused by maternal Parvovirus B19 infection].

Maternal infection with human Parvovirus B19 during pregnancy is one of the rare reasons, which can lead to a not immunologically conditioned hydrops fetalis. We report here a case of intrauterine Parvovirus infection associated with hydrops fetalis, diagnosed by intrauterine sonography in the 29 week of gestation, one week after the outbreak of Erythema. The B19 infection of the low birth weight baby, delivered in the 30 week of gestation, was diagnosed serologically with specific IgM and IgG against B19 by ELISA. No malformation could be detected in the liveborn infant, who died on the third day of life.

Antibodies, Viral↗
PubMed1989

Detection of parvovirus in a patient with "reactive arthritis" by in situ hybridization.

We used in situ hybridization to search for the presence of viruses in synovial fluid cell preparations obtained from patients with various forms of knee arthropathies. One patient, presenting with acute reactive arthritis, was found to replicate parvovirus DNA in cells from synovial fluid whereas six other patients with various other forms of arthritis were negative for parvovirus infection. Five patients with osteoarthritis, constituting a control group in which an infectious etiology would not be expected, were consistently negative. In addition, no hybridization was found when synovial cell preparations of all patients were hybridized in situ with DNA probes specific for Epstein-Barr virus, cytomegalovirus, or enteroviruses.

Adult↗