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Human parvovirus-associated arthritis: a clinical and laboratory description.

An outbreak of human parvovirus-associated erythema infectiosum in the Grampian region has enabled us to study the association between this small DNA virus and arthritis. This report deals with 42 patients with joint pains, in whom serological evidence of recent human parvovirus (HPV) infection was obtained. The clinical description of the disorder is based on 17 of the patients who were seen in the clinic and a further 13 patients for whom information was obtained by questionnaire. Detailed clinical features were not available for the remaining 12 patients. A rash was present in all 3 affected children but only 13 of the 27 adults. Viral prodromata were present in only 13 adults. 7 adults had neither rash nor viral prodromata. The arthritis was more common in adults than children and affected principally the female sex. In adults the arthritis was symmetrical, affecting the small joints of the hands, wrists, and knees most commonly. In all cases it was self-limiting, usually resolving within 4 weeks, although 1 adult had more persistent disease lasting almost 6 months. The 3 children reported in detail had less symmetrical and more persistent disease, which in 1 case has lasted over 7 months.

Adolescent↗

Human parvovirus and thalassaemia.

The human parvovirus (HPV) is responsible for aplastic crises in patients with chronic haemolytic anaemia. We describe the cases of four children with aplastic crises in various types of thalassaemia (alpha and beta thalassaemias, major and intermediate forms). In all four patients, specific anti-human parvovirus IgM was detected in their serum, thereby indicating recent infection.

Adolescent↗

Infection by human parvovirus B19: "gloves and socks" papular purpuric syndrome.

Human parvovirus B19 is the cause of erythema infectiosum a benign and self-limited infection, but sometimes the virus causes an acute and self-limiting dermatosis. It consists of a edema and erythema of the hands and feet in a gloves and sock distribution and is associated with oral lesions and fever. We report a case of a "gloves and socks" infection by human parvovirus B19.

Adult↗

Expression of the non-structural proteins of parvovirus MVMp from recombinant retroviruses: predominant role of the parvoviral NS-1 product in host cell disturbance.

Stable Psi-2 cell transformants were selected for their resistance to neomycin after transfection with a retroviral pZipNeo-SVX vector carrying sequences encoding for the non-structural proteins of parvovirus minute virus of mice (prototype strain, MVMp). Cells producing both NS-1 and NS-2 proteins (PsiNS) or only the NS-2 polypeptide (PsiNS2) were obtained. PsiNS cells exhibited morphological abnormalities and had a reduced clone-forming ability, whereas PsiNS2 cells were indistinguishable from the parental line. These cellular systems produced recombinant retroviral particles which transduced the NS gene(s) into mouse A9 cells. As in the case of Psi-2 cells, A9 transformants expressing both NS-1 and NS-2 proteins were impaired in their cloning efficiency. These results provided a direct confirmation of the predominant role of protein NS-1 in the cytopathic effect of parvoviruses.

Animals↗

Functional implications of the structure of the murine parvovirus, minute virus of mice.

BACKGROUND: Minute virus of mice (MVM) is a single-stranded (ss) DNA-containing, murine parvovirus with a capsid built up of 60 icosahedrally related polypeptide chains, each of which consists of the C-terminal region common to two structural proteins, VP1 and VP2. In infectious virions, most VP2 molecules are cleaved to VP3 by the removal of about 20 amino acids from the N terminus. Of the 587 amino acids in VP2, approximately half are identical to those in the analogous capsid protein of the antigenically distinct canine parvovirus (CPV), the crystal structure of which has previously been determined. The three-dimensional structure determination of MVMi (the immunosuppressive strain of MVM) was previously reported to 3.5 A resolution. RESULTS: We report here an analysis of the MVMi virus structure and provide insights into tissue tropism, antigenicity and DNA packaging. Amino acids determining MVM tissue tropism were found to cluster on, or near, the viral surface. A conserved, glycine-rich, N-terminal peptide was seen to run through a cylindrical channel along each fivefold axis and may have implications for antigenicity. Density within the virion was interpreted as 29 ssDNA nucleotides per icosahedral asymmetric unit, and accounts for over one-third of the viral genome. CONCLUSIONS: The presence of the glycine-rich sequence in the fivefold channels of MVMi provides a possible mechanism to explain how the unique N-terminal region of VP1 becomes externalized in infectious parvovirions. Residues that determine tropism may form an attachment recognition site for a secondary host-cell factor that modulates tissue specificity. The ordering of nucleotides in a similar region of the interior surface in the CPV and MVMi capsids suggests the existence of a genomic DNA-recognition site within the parvoviral capsid.

Amino Acid Sequence↗

Diagnostic assays with monoclonal antibodies for the human serum parvovirus-like virus (SPLV).

Monoclonal antibodies to the serum parvovirus-like virus (SPLV) were prepared by the hybridoma technique. They provided an antibody reagent which was used to develop solid phase antibody-capture assays for anti-SPLV IgM and IgG and for SPLV antigen. These assays were more sensitive than those based on human convalescent antibody as a reagent, and were more economical in the use of SPLV antigen. Their use enabled the serological responses to SPLV to be studied more fully and their sensitivity revealed the extent of SPLV infection. SPLV antigen was detected in four patients by both counter-immuno electrophoresis (CIE) and radioimmunoassay (RIA) and in two others by RIA alone. Parvovirus particles were seen in all six by electron microscopy. The anti-SPLV IgM response was measured in patients infected by SPLV. It was strong 5-18 days after the onset of illness, then declined and was only detectable in trace amounts after 6 months. Anti-SPLV IgG was also formed early, and persisted for at least 6 months. In a survey of 310 blood donors anti-SPLV was detected in 134 (43%) by CIE, but in 190 (61%) by IgG antibody capture RIA.

Antibodies, Monoclonal↗

Similarities in nucleotide sequence between serum and faecal human parvovirus DNA.

Two DNA clones were obtained from faecal specimens containing a parvovirus-like small round virus from a 1977 outbreak of gastroenteritis, and their nucleotide sequences were determined and found to be essentially identical with parts of the published sequence of serum parvovirus B19 and with a B19 isolate (JB) partially sequenced in this study. The clones corresponded mainly to genome regions coding for non-structural proteins, but also include a sequence of some 160 bp coding for structural proteins. Southern blotting experiments with a full-length B19 probe revealed a virion-sized 5 x 5 kbp DNA band in specimens from gastroenteritis cases in both 1977 and 1986. Thus the nucleotide sequence and hybridization results suggest that the virus seen in these studies is very similar to B19. Further work is necessary to clarify the antigenic relationship of these viruses.

Base Sequence↗

Incidental detection of parvovirus myocarditis at time of resection of discrete subaortic stenosis.

We report an asymptomatic 18-month-old boy who underwent surgical resection of the discrete subaortic stenosis. Histopathologic examination of the muscle demonstrated diffuse lymphocytic infiltration of the myocardium. Polymerase chain reaction analysis of blood demonstrated parvovirus. This report highlights that asymptomatic myocarditis may be more prevalent than we realise, and that involvement of the myocardium by parvovirus may be entirely asymptomatic.

Aortic Valve Stenosis↗

Expansion of tropism of a feline parvovirus to target a human tumor cell line by display of an alpha(v) integrin binding peptide on the capsid.

The autonomous parvoviruses are small, non-enveloped, single strand DNA viruses. They occur in many species and they have oncolytic properties. We are modifying the capsid of feline panleukopenia virus (FPV), a parvovirus which normally infects feline cells, with the goal of targeting human tumor cells for potential cancer therapy. Using recombinant viruses transducing a luciferase reporter, we show that insertion of a cyclically constrained, integrin-binding peptide at an exposed position on the FPV capsid enables transduction of an alpha(v) integrin-expressing human rhabdomyosarcoma cell line (Rh18A). These cells were not transduced by virus with the unmodified FPV capsid. Transduction of Rh18A was specifically inhibited by an alpha(v) integrin blocking antibody. However, other human tumor lines expressing alpha(v) integrins were not transduced by virus with either the modified or unmodified capsid. We conclude that modification of the FPV capsid to bind alpha(v) integrins can contribute to, but is not generally sufficient for, redirecting infection to human tumor cells. The permissiveness of Rh18A cells presumably involves additional factors unique to this line among various human cell lines tested.

Antigens, CD↗

Parvovirus-induced pancytopenia in a child with acquired haemolytic anaemia.

A 14-year-old boy, with acquired haemolytic anaemia secondary to paravalvular leak from a prosthetic aortic valve, presented with severe pancytopenia. Subsequent investigation showed serological evidence of recent parvovirus B19 infection. The patient required transfusion and subsequently improved rapidly, with maintenance of pre-infective haemoglobin levels. To our knowledge this is the first reported case in which infection with parvovirus has caused pancytopenia in an acquired chronic mechanical haemolytic anaemia.

Adolescent↗

Proteins tightly associated with the termini of replicative form DNA of Kilham rat virus, an autonomous parvovirus.

Revie et al. [Revie, D., Tseng, B. Y., Grafstrom, R. H. & Goulian, M. (1979) Proc. Natl. Acad. Sci. USA 76, 5539-5543] have proposed that the double-stranded replicative form (RF) DNA of the autonomous rodent parvovirus H-1 has protein of 60 kDa covalently bound at its 5' termini. We present evidence that the RF DNA of a similar rodent parvovirus, Kilham rat virus (KRV), also has covalently bound protein. NaDodSO4/polyacrylamide gel electrophoresis of purified, 125I-labeled RF DNA shows that proteins of 68-72, 66, 64, and 55 kDa copurify with the DNA during velocity and equilibrium sedimentation in the presence of detergents and 4 M guanidine HCl. Phenol extraction in the presence of 2-mercaptoethanol removes the 68- to 72-kDa proteins, but the 66-, 64-, and 55-kDa proteins remain tightly, but noncovalently, bound. The latter polypeptides also appear to associate with protease-treated RF DNA when mixed with uninfected cell extract. Following removal of these proteins by electrophoresis in NaDodSO4/agarose gels, two proteins (called RF TP-90 and RF TP-40), of about 90 and 40 kDa, become evident. These remain bound to the DNA and are released only after nuclease digestion of the DNA. These two proteins, apparently not of viral origin, are associated with terminal restriction fragments of the RF DNA and appear to be covalently bound to the 5' termini of both strands.

DNA Replication↗

Construction of a recombinant human parvovirus B19: adeno-associated virus 2 (AAV) DNA inverted terminal repeats are functional in an AAV-B19 hybrid virus.

To facilitate genetic analysis of the human pathogenic parvovirus B19, we constructed a hybrid B19 viral genome in which the defective B19 inverted terminal repeats were replaced with the full-length inverted terminal repeats from a nonpathogenic human parvovirus, the adeno-associated virus 2 (AAV). The hybrid AAV-B19 genome was rescued from a recombinant plasmid and then the DNA was replicated upon transfection into adenovirus 2-infected human KB cells in the presence of AAV genes coding for proteins required for AAV DNA replication (AAV-Rep proteins). In addition, in the presence of AAV genes coding for the viral capsid proteins (AAV-Cap proteins), the rescued/replicated hybrid AAV-B19 genomes were packed into mature AAV progeny virions, which were subsequently released into culture supernatants. The recombinant AAV-B19 progeny virions were infectious for normal human bone marrow cells and strongly suppressed erythropoiesis in vitro. The availability of an infectious recombinant B19 virus should facilitate the mutational analysis of the viral genome, which, in turn, may yield information on individual viral gene functions in B19-induced pathogenesis. The hybrid AAV-B19 genome may also prove to be a useful vector for gene transfer in human bone marrow cells.

Blotting, Southern↗

A model for tumor suppression using H-1 parvovirus.

A model system is proposed to investigate, at the molecular level, the pathways of tumor suppression. As a tool for the selection of cells with a suppressed phenotype, we used the H-1 parvovirus that preferentially kills various neoplastic cells. From the human K562 leukemia cells, we isolated a clone, KS, that is resistant to the cytopathic effect of the H-1 virus and displays a suppressed malignant phenotype. The suppressed malignancy and the cellular resistance to H-1 killing appear to depend on the activity of wild-type p53. Whereas the KS cells express wild-type p53, the protein is undetectable in the parental K562 cells. Experiments with p53 mutants suggest that wild-type p53, in its functionally intact state, contributes to the resistance against the cytopathic effect of H-1 parvovirus.

Antigens, CD↗

Canine parvovirus in New Zealand: epidemiological features and diagnostic methods.

Data from 763 cases of clinical canine parvovirus disease confirmed at the Ruakura Animal Health Laboratory were examined. The largest number of cases were seen in spring and summer months with the peak incidence in February 1981. The morbidity and mortality rates were highest in young dogs. Sixty-nine percent of all cases occurred in dogs less than six months of age, and 63 percent of dogs seven weeks of age or younger died. The laboratory methods used to diagnose canine parvovirus disease are compareh and discussed.

Journal Article↗

Porcine Parvovirus infection: review and diagnosis in a sow herd with reproductive failure.

In a commercial swine herd a rise was noted during the summer of 1981 in the number of repeat breeders, mostly four to eight weeks after serving. During the autumn there was a decrease in the litter size at birth and an increase in the number of stillborn and mummified piglets. Several gilts and sows showed a seroconversion against Porcine Parvovirus (PPV), determined by the Haemagglutination Inhibition test (HI-test). Characteristic pathological findings were seen in some maturely stillborn and neonatally decreased piglets (up to an age of 28 days); hepatic congestion and necrosis, accummulation of fluid in body cavities, myocarditis, and encephalitis were the most prominent features. Serological tests for antibodies in blood samples of one sow and body fluids of two stillborn piglets were suggestive of Porcine Parvovirus as the aetiological agent.

Animals↗

Production of recombinant H1 parvovirus stocks devoid of replication-competent viruses.

Vector and helper plasmids for the production of recombinant H1 (rH1) parvovirus, an oncolytic virus and candidate vector for cancer gene therapy, were constructed with the aim of reducing the contamination of these preparations with replication-competent viruses (RCV). Split-helper plasmids were constructed by manipulating the splicing signals for the capsid proteins such that VP1 and VP2 were expressed from separate plasmids. H1 vectors with similarly mutated splice sites were packaged, using the split-helper plasmids, and the resulting recombinant H1 viruses were completely free of RCV because the generation of recombinants expressing both capsid proteins was prevented. Vector yields of rH1 produced with split-helper plasmids in combination with splice site-modified vectors were similar (in the range of 10(7) replication units/ml) to yields of rH1 produced with the standard vector/helper pair, in which case significant levels of RCV were generated (10(4)-10(5) plaque-forming units/ml). To assess the functionality of this approach in vivo, rH1 was produced that contained the human interleukin 2 (IL-2) transgene and that was devoid of RCV. This IL-2-carrying rH1 vector expressed IL-2 efficiently in human tumor cells (HeLa) in vitro and generated antitumor responses in nude mice xenografted with HeLa cells that had been infected ex vivo with this virus. These results should allow the large-scale production of recombinant oncotropic parvoviruses and their assessment for the gene therapy of cancer in a clinical setting.

Animals↗

Recombinant LuIII autonomous parvovirus as a transient transducing vector for human cells.

Recombinants based on the genome of the autonomous parvovirus, LuIII, were constructed by replacing the viral coding sequences in an infectious clone (pGLu883) by a luciferase or beta-galactosidase reporter, which was linked to the viral P4 promoter. In cells cotransfected with either of these constructs, together with a plasmid supplying LuIII nonstructural and capsid proteins, excision and replication of the recombinant genome occurred. Transducing virions accumulated in the culture medium of the cotransfected cells, as assayed by reporter activity in recipient cells exposed to this medium. Transducing activity could be neutralized by antiserum to LuIII. Production of replicative form DNA and transducing virions were observed following cotransfection of HeLa, 293, or NB324K cells, in increasing order of efficiency. When homology existed between the recombinant genome and sequences flanking the viral genes in the helper construct, concomitant production of replication-competent, cytopathic virus was sometimes observed. This could be minimized by removal of the left end homology from the helper; by this means, preparations of luciferase transducing virus were obtained free from replication-competent virus. With such preparations, we observed luciferase expression (declining after 3 days) for up to 7 days in recipient HeLa cells. Hybridization of the recombinant viral DNA with strand-specific luciferase probes indicated packaging of both strands (as reported for LuIII), but with a several-fold excess of the (-) strand. We suggest that transducing-autonomous parvoviruses will be useful in gene transfer applications, possibly including gene therapy when only transient expression is desired.

Blotting, Southern↗

Deficient expression of enhanced reactivation of parvovirus H-1 in ataxia telangiectasia cells irradiated with X-rays or u.v. light.

Cells of patients with ataxia telangiectasia (AT), an inherited disease characterized by a high propensity to cancer, are hypersensitive to ionizing radiation. We investigated whether the hyper-radiosensitivity of AT cells correlated with a defect in their constitutive and/or conditional ability to rescue a damaged exogenous virus. For that purpose, parvovirus H-1, a single-stranded DNA virus whose intranuclear replication mostly relies on host cell functions, was used as a probe. The survival of u.v.- or gamma-irradiated H-1 was measured in X-, u.v.- or mock-irradiated human cells of normal (NB-E) or AT (AT5BIVA) origin. gamma-Irradiated H-1 survived to similar extents in untreated normal and AT cell lines. Both X- and u.v.-irradiation induced normal cells to achieve an enhanced reactivation (ER) of gamma- or u.v.-damaged H-1. In contrast, neither dose-effect curves nor time course revealed significant levels of ER expression after X- or u.v.-irradiation in AT5BIVA cells. Our results suggest that the impairment of ER of damaged parvoviruses may constitute a marker of the AT cell phenotype and be related to the radiosensitivity of AT cells.

Ataxia Telangiectasia↗