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A new parvovirus genotype persistent in human skin.

Parvovirus B19 is the exclusive human pathogen of the Erythrovirus genus. In classical view, the B19 DNA sequence shows little variability, with no disease-specific or tissue type specific associations. We examined skin biopsies from patients with B19-unrelated skin disease or from constitutionally healthy adults by polymerase chain reaction assays for four different genomic regions of the B19 virus. Sequencing showed that the skin-derived viral DNA differed within the protein-coding region from the B19 reference sequences by 10.8% and from the V9 variant by 8.6% and within the noncoding region (covering nucleotides 189-435 of the promoter region) by 26.5 and 17.2%, respectively. Despite this sequence difference, the promoter region was shown by a luciferase gene expression assay to be biologically active. We have detected a new B19 virus genotype, K71, which differs extensively from the known B19-virus genotypes and is persistently carried in human skin.

Biopsy↗

Intracellular localization of parvovirus B19 nucleic acid at the ultrastructural level by in situ hybridization with digoxigenin-labelled probes.

Conditions suitable for immunogold detection of digoxigenin-labelled DNA probes hybridized to parvovirus B19-infected erythroid cells embedded in Lowicryl K4M and LR White acrylic resins were established at the electron microscope level. The protocol was initially optimized using a positive control probe for whole human DNA which produced signal over the heterochromatin of all nucleated cells. In cultures harvested 2 days postinfection, B19 nucleic acid was detected mainly within the centrinuclear region of erythroid cells exhibiting characteristic margination of the chromatin. The B19 hybridization signal was largely unaffected by denaturation and was resistant to RNase digestion but sensitive to DNase digestion, indicating that it was mainly single-stranded B19 DNA. Relatively few gold particles were found over crystalline arrays of viral capsids, consistent with the observation that they are composed of mainly 'empty' capsids. B19 nucleic acid was detected in apparent transit from nucleus to cytoplasm through pores in the nuclear membrane. While the sensitivity of this system is limited by the fact that hybridization occurs only at the surface of the section, it is a rapid and specific means of localizing viral nucleic acids with a high degree of resolution.

Cells, Cultured↗

Combined immunocytochemistry and non-isotopic in situ hybridization for the ultrastructural investigation of human parvovirus B19 infection.

Parvovirus B19 is a single-stranded DNA virus with a specific tropism for human erythroid precursor cells. The virus codes for two overlapping structural (capsid) proteins and one non-structural protein which is thought to perform essential functions in viral replication, transcription and packaging. The ultrastructural localization of these proteins was achieved in cultured haemopoietic cells derived from fetal liver which had been infected in vitro and subsequently embedded in LR White acrylic resin. Postembedding immunogold detection of B19 structural and non-structural proteins was combined with localization of viral nucleic acid by in situ hybridization using a digoxigenin-labelled probe and different sized gold labels. The majority of the B19 capsid protein and DNA present in cells harvested 48 hours post-infection co-localized within the centri-nuclear region of erythroid cells demonstrating characteristic chromatin margination. Relatively little DNA hybridization signal was present over paracrystalline inclusions strongly labelled with anti-capsid protein monoclonal antibody R92F6. Viral DNA and capsid protein were co-localized in apparent egress from the nucleus through nuclear pores. B19 non-structural protein was detected in association with both nuclear and cytoplasmic arrays of capsids, supporting the view that this protein plays an important role in viral packaging and remains associated with the complete viral particle until its release from the cell. Co-localization of viral nucleic acid and proteins at the ultrastructural level is a flexible, rapid and highly specific tool for examination of viral life-cycles within cells.

Capsid↗

Prevalence of antihuman parvovirus B19 IgG antibodies in patients with refractory rheumatoid arthritis and polyarticular juvenile rheumatoid arthritis.

We investigated the prevalence of antihuman parvovirus B19 immunoglobulin G (IgG) antibody in 108 Japanese patients with rheumatoid arthritis (RA) and 11 patients with polyarticular juvenile rheumatoid arthritis (JRA). Seropositivity of anti-B19 was significantly higher in patients with refractory RA (57.6%, 38/66) compared with patients with remittent RA (19.0%, 8/42; P < 0.001) or age-matched controls (24.3%, 19/78; P < 0.001). Patients with refractory polyarticular JRA had a significantly higher frequency of anti-B19 seropositivity (71.4%, 5/7) than age-matched controls (8.3%, 5/60; P < 0.001), while none of the remittent group was positive for the antibody (0/4).

Adolescent↗

Persistence of rat parvovirus in athymic rats.

Euthymic (SD or outbred rnu/+) and athymic (rnu/rnu) rats were inoculated oronasally or intraperitoneally with the RV-Y strain of rat virus when they were 2 days or 4 weeks old. Clinical signs of infection in athymic infants were similar to those in euthymic infants, but significantly more athymic infants died. Some infants developed anemia and thrombocytopenia. After inoculation of infants. RV-Y was detected in surviving euthymic rats for 7 weeks and in surviving athymic rats for at least 10 weeks. After oronasal inoculation of 4 week-old rats no clinical illness was observed. RV-Y persisted less than 6 weeks in juvenile euthymic rats but at least 12 weeks in athymic juvenile rats. Intraperitoneal inoculation of juveniles resulted in infection for at least 6 weeks. The antibody response of athymic rats to RV-Y was significantly reduced compared to that of euthymic rats. These studies indicate that T cell deficiency increases the severity and duration of RV infection and imply that T cells are required for the full expression of resistance to RV infection. They also suggest that RV-Y induced anemia could serve as a model for human parvovirus-induced anemia.

Aging↗

Two anti-parvovirus B 19 IgM capture assays incorporating a mouse monoclonal antibody specific for B 19 viral capsid proteins VP 1 and VP 2.

During an outbreak of parvovirus B 19 in 1989 in Northern Ireland, 7580 blood donors were screened for B 19 antigen. Two units screened positive, one of which was obtained for use as viral antigen. A monoclonal antibody (R92F6) made against this antigen was specific for B 19 capsid proteins VP 1 and VP 2. The monoclonal antibody was used in the development of 2 anti-B 19 IgM capture enzyme assays. These used a conventional substrate (O-phenylene diamine) and a chemiluminescent signal reagent. There was excellent concordance between the 2 assays. A total of 403 patients sera were tested and 65 sera were positive in each assay.

Antibodies, Monoclonal↗

Sequence analysis of a parvovirus B19 isolate and baculovirus expression of the non-structural protein.

Serology for parvovirus B19 has been hampered by limited availability of antigen which has often had to be isolated from viraemic blood donations. We have determined the sequence of the genome of one such isolate (Stu). It is 99% similar to the sequences of two other isolates (Wi and Au) except at the far 5'-end, where it is more similar to the terminus of another isolate (Ala/Alb). Recombinant nonstructural protein, NS, was constructed. Antibodies to NS, as well as to the capsid proteins, VP1/2, were detected in patients with B19 infection.

Animals↗

Persistent human parvovirus B19 infection following an acute infection with meningitis in an immunocompetent patient.

A case in which parvovirus B19 infection persisted over a prolonged period of time in the blood of an immunocompetent patient following an acute infection with meningitis is reported. Using a nested polymerase chain reaction assay the viral genome was detected in cerebrospinal fluid as well as in blood at the time of overt disease and in consecutive blood samples collected for up to nine months.

Adult↗

Human parvovirus B19 and rheumatoid arthritis.

Human parvovirus B19 infections have been linked with the development of a short-lived symmetrical polyarthritis and, rarely, a more persistent arthritis. We prospectively looked for serological evidence of recent B19 infection in 25 early synovitis patients presenting within 12 weeks of symptom onset and compared them with 21 controls seen over the same time period. None of the control patients had evidence of recent B19 infection while 3 of the early synovitis patients had raised IgM anti-B19 antibody levels. Two had a transient arthritis and 1 developed persistent seropositive rheumatoid arthritis.

Adult↗

[[Human parvovirus B19 and its clinical significance].

Human parvovirus B19 causes erythema infectiosum (fifth disease) and aplastic crisis. In pregnancy, B19 may be transmitted on to the fetus resulting in hydrops fetalis. B19 is sometimes associated with thrombocytopenia and vasculitis. In immunocompromised patients, B19 may persist causing chronic infection. Several methods for detecting B19 virus or specific antibodies are now available. B19 infection does not usually require therapy. Efficacy of immunoglobulins in chronic B19 infection has been demonstrated.

Antibodies, Viral↗

Three cases of myocarditis in childhood associated with human parvovirus (B19 virus).

We report three childhood cases of myocarditis associated with human parvovirus (B19 virus). All three children presented with significant cardiac decompensation, with one requiring extracorporeal membrane oxygenation support. Left ventricular function was severely impaired in all three. Myocardial biopsy confirmed histological myocarditis and was positive for B19 virus by nested polymerase chain reaction. Serum was positive for IgG B19 virus but negative for IgM in all three cases. All three children were treated with diuretics, ACE inhibitors, and immunosuppression. Prednisone and cyclosporin were continued until there was echocardiographic and histological improvement. All made a full clinical and echocardiographic recovery.

Child↗

Severe aplastic anemia associated with human parvovirus B19 infection in a patient without underlying disease.

Human parvovirus B19 (B19 virus) infection is known to induce aplastic crisis in patients with hemolytic anemia. In healthy subjects, B19 infection may sometimes cause mild pancytopenia, but these changes are transient and recovery is spontaneous. We report the first case of aplastic anemia in a previously healthy boy without any underlying diseases, following asymptomatic infection with the B19 virus. Laboratory examination initially showed thrombocytopenia, mild leukopenia in the peripheral blood, and severe hypoplastic bone marrow. Since pancytopenia developed and worsened progressively, immunosuppressive therapy was given, resulting in a complete remission. Despite the lack of an infectious prodrome, serological and histological analysis revealed an underlying infection with the B19 virus. Thus, B19 virus infection must be considered one of the causes of aplastic anemia in patients without underlying hemolytic anemia and an apparent episode of the viral infection.

Adolescent↗

Parvovirus B19 associated neutropenia. Treatment with Rh G-CSF.

In a 39-year-old patient under follow-up for metastatic adenocarcinoma of the colon, a post-operative fever and a profound neutropenia occurred during hospitalization. A diagnosis of primary infection with parvovirus B-19 was made in view of the association of reticulocytopenia, together with anti-B19 IgM antibodies and positive serum results for viral DNA. The granulocytic lineage appeared to be normal on bone-marrow smears, and anti PMN autoantibodies were not found. Rh G-CSF therapy resulted in a sharp increase in the PMN count. Although the pathophysiology of B19-associated neutropenia remains unclear, Rh G-CSF seems to be effective in this disorder.

Adenocarcinoma↗

Parvovirus [correction of Parovirus] B19-induced red cell aplasia in solid-organ transplant recipients. Two case reports and review of the literature.

Two solid-organ transplant recipients (one heart and one lung) developed severe anemia with reticulocytopenia. Both were heavily immunosuppressed. Bone marrow aspiration revealed almost complete absence of erythroid precursors. A few giant megaloblastic proerythroblasts with cytoplasmic vacuolisation and intranuclear inclusions were seen. Human parvovirus B19 (B19V)-DNA genome was found by nested-PCR assays in blood and bone marrow samples in both cases. Twelve similar cases are described in the literature. When looked for, B19V DNA was positive either in serum or bone marrow or both. Twelve of the fourteen patients were successfully treated by high dose i.v. immunoglobulin (IVIG). One patient recovered spontaneously and another after treatment with recombinant human erythropoietin (rHu-EPO) only. Transplant patients should be considered at risk for severe erythroblastopenic anemia due to B19V infection. Diagnosis is based on bone marrow examination and detection of B19V DNA by PCR in serum and/or marrow. IVIG is an effective and safe treatment. The role of erythropoietin in this indication needs further study.

Adult↗

The relationship between arthritis and human parvovirus B19 infection.

In order to evaluate the role of human parvovirus B19 in the etiopathogenesis of autoimmune diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), synovial fluid and blood specimens were collected at 1-month intervals from 20 patients with early synovitis (ES) and 31 with RA. Blood specimens were also collected from 25 patients with SLE, 25 with osteoarthritis (OA) as the diseased control group, and 50 healthy blood donors (HBD) as the healthy control group. Detection of B19 IgM and B19 IgG were performed by enzyme-linked immunosorbent assay from serum specimens, and B19 DNA was detected by polymerase chain reaction from synovial fluid samples. B19 IgM, B19 IgG, and B19 DNA were found in the three patients of the ES group. Subsequently, two of them were diagnosed with RA and one with SLE. B19 DNA was also detected in the synovial fluid of eight patients in the RA group. Of them, all were positive for B19 IgG and half were positive for B19 IgM. B19 IgM was not detected in either of the control groups. To define the role of B19 in the etiopathogenesis and prognosis of undiagnosed arthritis and other chronic inflammatory diseases such as RA and SLE, we need broader serial and prospective studies based on clinical and laboratory collaboration. In conjunction with case reports, these studies would also serve to detect other possible factors in the etiopathogenesis of chronic inflammatory diseases.

Adult↗

A case of pure red cell aplasia and systemic lupus erythematosus caused by human parvovirus B19 infection.

Human parvovirus B19 (B19) rarely induces pure red cell aplasia (PRCA) in healthy hosts. Meanwhile B19 infection is often clinically similar to systemic lupus erythematosus (SLE), and several cases have been reported wherein B19 actually stimulated SLE exacerbation in an immunocompetent subject. An 82-year-old healthy woman was diagnosed to have complicated with B19 infection and PRCA. Four weeks later, she had high fever, polyarthritis, and oral ulcers, additionally diagnosed with SLE, and subsequently, 15 mg of prednisone was started. This is the first case wherein B19 infection caused both PRCA and SLE in a healthy patient as far as our investigations are concerned.

Aged, 80 and over↗

A high prevalence of parvovirus B19 DNA in patients with psoriasis.

Psoriasis is a common inflammatory skin disease. Infectious models are considered to be of pathophysiological importance in psoriasis. The immunological profile of stable psoriasis plaques suggests that viral antigens may be important. Human parvovirus B19 (PVB19) is a single-stranded DNA virus that causes various clinical symptoms. Several case reports have suggested associations between PVB19 infection and various chronic autoimmune and dermatologic diseases. There has so far been no information regarding the role of PVB19 in psoriasis, except psoriatic arthritis. In this report, to investigate the role of PVB19 in psoriasis, we analyzed PVB19 DNA of peripheral blood from psoriatic patients (n = 47) in comparison with blood donors (n = 20). We also determined the presence of anti-PVB19 IgG and IgM antibodies by using enzyme-linked immunosorbent assay (ELISA). We found that the presence of PVB19 DNA in patients with psoriasis (38%) was significantly higher than in controls (0%, P < 0.01). Anti-PVB19 IgG antibodies were detected in 79% of the cases while only 6% had anti-PVB19 IgM antibodies. PVB19 DNA presence was associated with seropositivity for anti-PVB19 IgG (P < 0.05) but not with IgM antibodies, indicating subclinical activation of latent infection. No correlation was found between the presence of PVB19 DNA and a patient's age, sex, type of psoriasis, or psoriasis area and severity index. The data demonstrated a statistically significant association between psoriasis and PVB19. Therefore, we suggest that PVB19 infection may be of pathophysiological importance in psoriasis.

Adolescent↗

Parvovirus B19: a pathogen responsible for more than hematologic disorders.

The clinical and pathomorphological patterns of parvovirus B19 (PVB19)-associated diseases is the result of a balance between virus, host target cells and immune response. It is a characteristic feature of PVB19 that in patients with various other preexisting diseases, e.g., many hemolytic anemias, immune complex-mediated vasculitic disorders, and primary or secondary immunodeficiencies, the underlying diseases can be triggered, aggravated or complicated by severe organ manifestations. Identification of PVB19 by means of routine histology and immunohistology is only given in lytic infections occurring in transient aplastic anemia or nonimmune hydrops fetalis by the detection of viral inclusion bodies in erythroid precursor cells. In all other PVB19-associated diseases, molecular pathological methods must be applied. In this report, quantitative real-time polymerase chain reaction was used to determine the viral load in formalin-fixed and paraffin-embedded tissues derived from various organs. Using in situ hybridization it was demonstrated that endothelial cells of the microcirculatory periphery of the heart and hepatobiliar system in lytic infections are PVB19-specific target cells in children and adults. Because treatment of lytic PVB19 infection has been successfully applied, the pathologist should be alerted to include PVB19 into the diagnostic spectrum of viral disease, especially in immunocompromised patients.

Adult↗