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PubMed1992

In vitro culture for the detection of infectious human parvovirus B19 and B19-specific antibodies using foetal haematopoietic precursor cells.

The inability to culture human parvovirus B19 in standard cell lines has rendered investigation of clinical samples for the presence of infectious virus problematic. Using haematopoietic precursors derived from first trimester foetal liver as targets for infection, and non-isotopic in situ hybridization to detect intracellular viral DNA, we have assessed infectivity in stored serum samples taken from nine volunteers at different stages following intranasal inoculation with parvovirus B19. Infectious virus was detected as early as 3 days after inoculation, the cessation of infectivity correlating with the rise in specific IgM. In all but two samples, infectivity correlated with the detection of B19 DNA by dot-blot hybridization, although in vitro culture was 10-fold more sensitive than dot-blot hybridization. B19 DNA was detected by the polymerase chain reaction in serum from one volunteer up to 36 days after inoculation, although samples containing specific antibody were non-infectious. Infection of erythroid precursors was completely inhibited by preincubation of virus with serum containing high titre B19-specific IgM and IgG. Unexpectedly, this was associated with a strong B19 DNA hybridization signal within the cytoplasm of phagocytic macrophages. This culture and detection system is a rapid and sensitive means of detecting infectious virus in serum samples, and of assessing the neutralizing ability of B19-specific antibodies.

Antibodies, Viral↗
PubMed1992

Non-permissiveness of synovial membrane cells to human parvovirus B19 in vitro.

The ability of cultured human synovial cells derived from synovial membrane and cartilage to support the replication of human parvovirus B19 was assessed. No viral DNA synthesis nor viral antigens were detected suggesting that B19 virus is not capable of replicating in synovial cells. The significance of this finding in relationship to the pathogenesis of parvovirus arthritis is discussed.

Bone Marrow↗
PubMed1992

Haemagglutination by parvovirus B19.

Human parvovirus B19 is a member of the autonomous parvoviridae but in contrast to other members of the genus has not been shown to agglutinate red blood cells. We now report that the virus agglutinates red cells of primate origin, though with plasma-derived virus this activity is masked by the presence of an IgM-like inhibitor. This observation is consistent with the presence on the erythroid precursor target cell of a specific receptor for parvovirus B19.

Animals↗
PubMed1991

Chemiluminescence dot blot hybridization assay for detection of B19 parvovirus DNA in human sera.

A chemiluminescence dot blot hybridization assay was used for the detection of B19 parvovirus DNA in human sera by using digoxigenin-labeled probes. The probes were revealed immunoenzymatically by use of anti-digoxigenin Fab fragments conjugated with alkaline phosphatase. The chemiluminescence signal was obtained by reacting the labeled probe-target complex with an enzyme-triggerable dioxetane substrate. The emitted photons were detected with instant photographic films. In the search for B19 parvovirus DNA, 2,808 serum samples were analyzed.

DNA Probes↗
PubMed1992

Prokaryotic expression of a VP1 polypeptide antigen for diagnosis by a human parvovirus B19 antibody enzyme immunoassay.

To produce parvovirus B19 antigen for diagnostic purposes, partially overlapping segments covering the genes encoding the viral structural proteins VP1 and VP2 were cloned into expression vectors. The constructs were induced in Escherichia coli, resulting in the expression of beta-galactosidase fusion proteins. In immunoblotting experiments with sera from patients with erythema infectiosum, immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide of 235 amino acids at the N terminus of VP1. The DNA fragment encoding this polypeptide was amplified by the polymerase chain reaction and cloned into an expression vector. The viral capsid antigen expressed in E. coli was purified by preparative agarose gel electrophoresis and used in IgG and IgM solid-phase enzyme immunoassays. Comparison with reference gamma- and mu-capture radioimmunoassays using whole virus antigen showed that these antibody tests are suitable for the serodiagnosis of human infections caused by parvovirus B19.

Antibodies, Viral↗
PubMed1992

Possible induction of systemic lupus erythematosus by human parvovirus.

A 59 year old woman presented with an influenza-like illness preceding signs and symptoms strongly suggestive of systemic lupus erythematosus (SLE), which progressed over several months. Owing to these influenza-like symptoms, a viral cause of her illness was sought. Human parvovirus B19 serology was positive and antibodies to DNA were detected by two different methods. This patient is believed to be the first report of human parvovirus B19 infection coinciding with the onset of SLE. The evidence for B19 virus and the part it plays in autoimmunity and arthritis is discussed.

Antibodies, Antinuclear↗
PubMed1992

Non-isotopic in situ hybridisation and immunophenotyping of infected cells in the investigation of human fetal parvovirus infection.

AIMS: To compare the use of biotinylated and digoxigenin labelled probes for diagnosis of human fetal parvovirus B19 infection in formalin fixed, paraffin wax embedded tissues; and to assess the cellular distribution of the virus in positive cases. METHODS: Sections of lung tissue from 23 cases of anatomically normal non-immune fetal hydrops presenting between 1984 and 1989, and from 13 control cases of hydrops due to chromosomal abnormality were probed for B19 DNA by in situ hybridisation using both biotinylated and digoxigenin labelled probes. The distribution of the virus was then investigated in all cases of fetal B19 infection confirmed in this laboratory to date (n = 11) by combining in situ hybridisation for viral DNA (using the digoxigenin system) with immunohistological labelling for a range of cellular antigens. RESULTS: Five unequivocal cases of B19 infection were identified among the 23 fetuses with unexplained hydrops using both probe labels. When combined with data from previous studies of the period 1974-1983, the results indicate that B19 infection was responsible for 27% of cases of anatomically normal non-immune hydrops and 8% of all cases, of non-immune hydrops presenting to this hospital over 15 years. False positive signal was seen in an additional three cases, using biotinylated probes. Digoxigenin labelled probes gave greater specificity and permitted detailed investigation of tissues high in endogenous biotin. Though most cells containing B19 DNA colabelled as erythroid precursors, viral DNA was frequently detected within mononuclear-phagocytic cells. In three cases viral signal was also found within occasional myocardial cells labelled by antibody to desmin. CONCLUSIONS: A relatively high proportion of cases of anatomically normal, non-immune hydrops are caused by B19 infection. Digoxigenin is a more reliable probe label than biotin for in situ hybridisation in archival fetal tissues. Double labelling for cellular antigens and viral nucleic acid is a powerful technique for investigating virus-host cell interactions, and provides evidence that cell types other than those of erythroid lineage may have a role in human fetal parvovirus infection.

DNA Probes↗
PubMed1991

Parvovirus B19 infection can induce histiocytic necrotizing lymphadenitis (Kikuchi's disease) associated with systemic lupus erythematosus.

We report three cases of systemic lupus erythematosus (SLE) associated with necrotizing histiocytic lymphadenitis (Kikuchi's disease) and immunologically proven human parvovirus B19 infection. Simultaneous occurrence of SLE and Kikuchi's disease was a characteristic of the three cases. Kikuchi's disease is an uncommon disease that usually affects young women and is characterized by painless unilateral cervical lymph-node enlargement. T-cell regions of affected lymph nodes are exclusively involved with patchy paracortical necrosis surrounded by a polymorphous cell population of histiocytes and macrophages. However, lymphadenopathy in patients with SLE may be histologically indistinguishable from Kikuchi's necrotizing lymphadenitis. The cause of Kikuchi's disease remains uncertain, although infectious agents have been proposed. A positive IgM-specific anti-human parvovirus B19 antibody test in our three cases suggests that B19 can induce a necrotizing histiocytic lymphadenitis and possibly a clinical SLE flare. High-dose (1 mg/kg/day) and medium-dose (0.5 mg/kg/day) oral prednisone was an effective treatment for constitutional and visceral symptoms of Kikuchi's and SLE diseases.

Adolescent↗
PubMed1992

Polymerase chain reaction with double primer pairs for detection of human parvovirus B19 induced aplastic crises in family outbreaks.

Parvovirus B19 DNA can be detected by polymerase chain reaction with double primer pairs (nested PCR). Recent infection was documented by a retrospective serological study using Parvoscan-B19 enzyme linked immunosorbent assay (EIA) for detection of B19 human parvovirus IgM and IgG antibodies in serum or plasma specimens. In 3 families B19 outbreaks caused aplastic crises necessitating blood transfusion in 5 children and 1 adult with hereditary sphaerocytosis. Four members from 2 of the families had clinically overt haemolytic anaemia prior to the event. Two members in another family presented with an aplastic crisis disclosing the underlying chronic haemolytic disease. All 7 patients were identified as PCR positive in serum samples taken 3-14 days after the onset of symptoms. Comparison with dot blot hybridization revealed detectable DNA in only 2/3 PCR positive patients. Thus, nested PCR is more sensitive than the dot blot hybridization method and is therefore a suitable complement to the antibody assay for identifying recent B19 infection.

Adolescent↗
PubMed1992

Bone marrow necrosis and human parvovirus associated infection preceding an Ph1+ acute lymphoblastic leukemia.

A case of bone marrow necrosis associated with a serologically documented recent Parvovirus B 19 infection which preceded the development of PH1+ acute lymphoblastic leukemia is reported. No conclusions can be drawn on the basis of a single case but the question of the role of human Parvovirus B19 in the pathogenesis of bone marrow necrosis is discussed. It is suggested that the virus may act as a co-factor for the induction of bone marrow necrosis, in some cases.

Adult↗
PubMed1991

[Human parvovirus B19 as the cause of aplastic crisis in hereditary spherocytosis].

A seven months old boy was admitted to hospital for investigation and treatment of severe anaemia. The final diagnosis was hereditary spherocytosis. When, six weeks later, he developed an aplastic crisis, serological criteria provided evidence of recent human parvovirus B19 infection. The same disease was detected in the family. Human parvovirus B19 is shown to be of particular interest in aplastic crisis.

Anemia, Aplastic↗
PubMed1992

First continuous propagation of B19 parvovirus in a cell line.

The pathogenic human parvovirus B19 has extreme tropism for human erythroid progenitor cells and has resisted cultivation in conventional cell lines. We report first propagation of this virus in an erythropoietin-dependent strain of a megakaryoblastic leukemia cell line called UT-7. Virus protein was present in about 5% of cells after 1 week of culture. Appropriate ratios of major and minor capsid proteins were determined by immunoblot, and newly synthesized capsid protein was detected by immunoprecipitation of radioactively labeled cell lysates. High molecular weight monomer and dimer intermediates were detected by Southern analysis, indicating active viral replication. Approximately 1,000 genome copies were present per infected cell, and at the optimal multiplicity of infection 20- to 50-fold more virus was produced than inoculated. Virus propagation only occurred in UT-7 cells that were adapted to growth in erythropoietin; virus signal was not detected in UT-7 cells adapted for growth in granulocyte-macrophage colony-stimulating factor or interleukin-3, even with exposure to erythropoietin for several days. Infectious virus was detected in cultures as long as 3 months after inoculation. Despite persistence, there was no evidence of viral integration on Southern analysis. This cell line may prove useful for the production of infectious virus and in the analysis of B19 parvovirus persistence, cytotoxicity, and permissivity.

Blotting, Southern↗
PubMed1991

[Human Parvovirus B19--really only fifth disease? Unusual disease course in children and adolescents].

The human parvovirus B19 agent causes infectious erythema (fifth disease). However, a wide range of other pathological manifestations may also be seen: atypical exanthema, ARD (also obstructive forms, e.g. bronchiolitis), acute gastroenteritis, chronic anemia or aplastic crises (in constitutional or malignant hematological diseases or immunological deficiency), arthralgia/arthritis (e.g. rheumatoid arthritis, jcA), diseases of the central nervous systems (e.g. febrile convulsions in young children), lymphadenopathies (e.g. lymphadenitis mesenterialis or pseudoappendicitis); prenatal infection can lead to fetal death (not malformations!). Infection occurring concomitantly with vaccination may suggest complications of the latter. To clarify the true etiological situation, modern laboratory investigations are then required. Vaccination against parvovirus B19 (initially indicated in the case of non-immune girls and women wanting children) is a desirable future development.

Adolescent↗
PubMed1978

H-1 and X14 parvovirus antibodies in women with abortions or still-births.

Antibodies against H-1 and X14 parvoviruses were found 13.71 and 9.14%, respectively, of 350 sera from women affected by repeated abortions or still-births. On the contrary, only 2.66 and 1.66% of 300 control sera were positive to the two viruses. The difference in the incidence of antibodies appears highly significant (p greater than 0.001). Therefore a possible role of the parvoviruses in inducing abortions or still-births in humans is suggested.

Abortion, Spontaneous↗
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↗
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↗