Fetal brain infection with human parvovirus B19.

Intrauterine parvovirus B19 infection is known to be one of the causes of hydrops fetalis. However, there are few reports of the pathologic changes in the central nervous system. Postmortem examination of a fetus revealed multinucleated giant cells of macrophage/microglia lineage and many small calcifications around the vessels, predominantly in the cerebral white matter. Parvovirus B19 genome DNA was detected in the nucleus of the multinucleated giant cells and solitary endothelial cells by polymerase chain reaction amplification and in situ polymerase chain reaction methods. Capsid antigen was also demonstrated in the cytoplasm of the endothelial cells by immunofluorescent assay. Thus, intrauterine B19 parvovirus infection could be associated with marked neuropathologic changes in the fetal brain at the midembryonal period. Neurologic follow-up of complications may be necessary for children who survive the intrauterine infection.

Acute cerebellar ataxia with human parvovirus B19 infection.

A 2 year old boy developed acute cerebellar ataxia in association with erythema infectiosum. During the disease, genomic DNA and antibodies against human parvovirus B19 were detected in serum but not in cerebrospinal fluid. Parvovirus B19 associated acute cerebellar ataxia might occur due to transient vascular reaction in the cerebellum during infection.

A case of prolonged human parvovirus B19 DNA-emia associated with polyclonal B cell activation.

The current paper reports an 8 year old girl with arthralgia and polyclonal B cell activation induced by human parvovirus B19 infection (HPV B19). The infection was diagnosed by the presence of the virus genome in sera. The patient presented with transient arthritis in the wrist, ankle joint and neck and elevation of immunoglobulin IgM antibodies to HPV B19 and rubella, antibodies to Mycoplasma and antistreptolysin O but without the typical clinical features of erythema infectiosum. The polyclonal B cell activation was paralleled by the presence of the virus genome of HPV B19 in sera. In some children with arthralgia, it is important to examine the genomes of viruses that may cause arthritis as well as the antibody titers to the viruses.

A new genome type of human parvovirus B19 present in sera of patients with encephalopathy.

The possible involvement of human parvovirus B19 infection in encephalopathy has been reported. To determine the characteristics of B19 viruses involved in such cases, we molecularly cloned a part of the B19 DNAs derived from the sera of three patients with encephalopathy (B19 strains N80, N81 and N82), following amplification by PCR. The nucleotide (nt) sequences of the cloned DNAs (nt 3147-3405) were then determined. The nucleotide sequence of N80 was similar to that of the known genome type of group II. The nucleotide sequences of N81 and N82 were similar to each other, but distinctly different from those of other B19 strains reported previously. Almost the entire genome of strain N81 (nt 229-4812) was molecularly cloned following amplification by PCR. Analyses of the restriction site polymorphisms of the cloned N81 DNAs showed that N81 is distantly related to other B19 strains. Therefore, the two strains of N81 and N82 were defined as belonging to a new genome type, group V. Group V B19 virus has so far been isolated only from patients with encephalopathy.

Genetic diversity in the non-structural gene of parvovirus B19 detected by single-stranded conformational polymorphism assay (SSCP) and partial nucleotide sequencing.

A homologous region in the parvovirus B19 non-structural gene (B19 nt 1399-1682) was examined in 50 samples from patients with a wide variety of B19-related disease from various countries by PCR amplification, single-stranded conformational polymorphism (SSCP) assay and nucleotide sequencing. Five SSCP types were confirmed by nucleotide sequence analysis. Of a total of 6 mutations, all were silent. Types 3 and 4 accounted for 92% of strains. There was no correlation between genome type and either clinical illness or patient age. However, there was a correlation between SSCP type and country of origin. Type 3 strains predominated in Japan (18/26) and the UK (6/8), whereas type 4 predominated in the USA (9/12). Notably, type 3 strains also predominated among females (14/18), whereas there were approximately equal numbers of strain types 3 (7/17) and 4 (8/17) among males; an observation which remains unexplained. Within the Japanese group, although type 3 strains predominated overall, strains isolated from 1981 to 1987 consisted of types 1 (2/15), 2 (1/15), 3 (8/15), and 4 (4/15), whereas strains isolated from 1990 to 1994 consisted almost entirely of type 3 (10/11).

[Responsiveness of a new parvovirus B19 antigen produced in baculovirus expression system to B19-specific IgG and IgM antibodies in every epidemic, 1968, 1980, 1987 and 1992 in Japan].

A new recombinant parvovirus B19 antigen was tested whether it was responsive to human serum antibodies in every epidemic year of erythema infectiosum for 25 years, because wild strains of B19 parvovirus were changeable genetically. The antigen was empty particles of both B19-VP1 and VP2 produced in baculovirus expression system. Specimens were 21 sera in 1968, 19 in 1980, 44 in 1987 and 33 in 1992, derived from 67 patients with erythema infectiosum, fever and/or non-specific exanthem and aplastic crisis in persons with hereditary spherocytosis. Each patient had been confirmed of B19 parvovirus infection by other methods as radio immunoassay and/or enzyme-linked immunosolvent assay for B19-IgG and IgM using other antigens and by detection of B19-genome DNA using the polymerase chain reaction. Days of the illness of every serum were confirmed including before infection to 216 days after onset. Sera from 23 patients with measles, Kawasaki disease and rubella were selected for controls, and those patients who had not been infected with B19 parvovirus. Tests were carried out by enzyme immunoassay, indirect method for IgG and IgM capture method. In a total of 103 specimens after onset of symptoms B19-IgG was positive in yearly specimens, and B19-IgM was also positive in all acute phase sera. B19-IgG in most of all sera was kept in peak level up to 216 days after onset. B19-IgM increased rapidly in acute phase and seemed to disappear within one to 5 months after onset. Thirty-seven specimens including 14 obtained at state before infection and 23 controls were completely negative for both B19-IgG and IgM.(ABSTRACT TRUNCATED AT 250 WORDS)

Partial nucleotide sequencing and characterization of human parvovirus B19 genome DNAs from damaged human fetuses and from patients with leukemia.

While human parvovirus B19 is associated with fetal damage and chronic suppression of bone marrow in patients with leukemia, much less is known of the genomic characteristics of B19 isolated from damaged human fetuses and leukemia patients. B19 genome DNAs from human fetal organs and fluids and sera from leukemia patients were amplified by the polymerase chain reaction. The nucleotide sequences of amplified products in the region between nucleotide (nt) 3141 and nt3411 were determined following molecular cloning in M13 phage. The genome types of B19 from the fetuses and leukemia patients were similar to the types from patients with aplastic crisis and an asymptomatic individual. Wide diversity of the nucleotide sequence, i.e., six or more (6-11) substitutions, were evident in ten M13 phage clones of each DNA source from fetal materials, while substitutions in sera from patients with leukemia and aplastic crisis and of an asymptomatic individual numbered four or fewer (0-4). This wide diversity of B19 viruses in the fetus, revealed after many rounds of DNA replication, probably depends on a persistent state of infection.

Genetic diversity of human parvovirus B19 determined using a set of restriction endonucleases recognizing four or five base pairs and partial nucleotide sequencing: use of sequence variability in virus classification.

Analysis of the restriction site polymorphism (RSP) of human parvovirus B19 using 12 restriction endonucleases (REs) recognizing four or five bp sequences (4- or 5-bp REs) revealed a significant difference between strains previously classified as being of the same genome type, and a relationship between two strains of different genome types, thereby indicating a global spread of B19 virus strains. These findings demonstrate the advantage of this set of 4- and 5-bp REs for the calculation of the degree of genetic diversity and clearly it is necessary to amend the taxonomy of B19 virus strains using these REs. We examined the nucleotide (nt) sequence between nt 3141 and 3411, at the N terminus of the VP2 protein coding region, in 12 B19 virus strains. The pattern of distribution of nucleotide differences between the strains confirmed the classification by RSP analysis. Between nt 3293 and nt 3364, a region in which an antigenic epitope may be encoded, there was no evidence of a nucleotide change causing an amino acid change. Thus, the amino acid sequence in this potential epitope is probably conserved.

The genome type of human parvovirus B19 strains isolated in Japan during 1981 differs from types detected in 1986 to 1987: a correlation between genome type and prevalence.

The genome DNAs of 12 strains of human parvovirus B19, isolated in Japan at two different times, 1981 and 1986 to 1987, were molecularly cloned in the plasmid pUC18. The cloned B19 DNAs were analysed by cleaving with restriction endonucleases, and were classified into several groups by genome type. The restriction endonuclease cutting patterns of B19 strains isolated during 1981 were similar to that of the group IV genome type, and the patterns of those isolated later were similar to that of group II, suggesting a correlation between the genome type and the prevalence. We conclude that the prevalences of B19 infection in Japan during 1981 and in 1986 to 1987 were caused by viruses differing in genome type, and that B19 viruses with similar genome types disseminated widely in Japan during each prevalence.

[Aplastic crisis due to human parvovirus B19 infection in glucose-6-phosphate dehydrogenase deficiency].

Human parvovirus B19 is known to cause aplastic crisis in patients with hemolytic anemias due to cytotoxic effect of the infection to erythroid progenitor cells. We report here the first case of aplastic crisis by B19 in a patient with glucose-6-phosphate dehydrogenase deficiency. A five-year-old boy was admitted to the hospital because of severe anemia, fever and jaundice. Four weeks after admission, he developed erythema infectiosum. B19 infection was confirmed using countercurrent immunoelectrophoresis, Southern blotting and hybridization method, and radioimmunoassay for B19 specific IgM. B19 virus antigen was detected by an indirect immunofluorescent method in both the cytoplasm and nucleus of large mononuclear cells that had no granules in bone marrow. On admission, the hemoglobin was 3.1 g/dl and no reticulocytes were detected in the peripheral blood smear. Bone marrow examination revealed a normocellular marrow with erythroid hypoplasia and M/E ratio of 7.38. Large basophilic erythroblasts containing vacuoles were also noticed. Elevation of indirect bilirubin and hemoglobinuria suggested intravascular hemolysis. Transient mild thrombocytopenia associated with increased PAIgG was observed. It is likely that B19 virus infection caused hemolysis which contributed to severe anemia.

Nonimmunologic hydrops fetalis resulting from intrauterine human parvovirus B-19 infection: report of two cases.

Two cases of nonimmunologic hydrops fetalis associated with intrauterine human parvovirus B-19 infection are reported. In these cases, hydrops fetalis was diagnosed with ultrasound at 21 and 22 weeks' gestation after 10 or more days of maternal flu-like symptoms. The outcome was stillbirth in one case and neonatal death in the other. In both cases, intrauterine infection by human parvovirus B-19 was confirmed based on two findings: maternal serum positive for human parvovirus B-19 immunoglobulin M (IgM) antibody, and human parvovirus B-19 DNA detected in the fetal organs using Southern blotting and hybridization with a 32P-labeled probe. Laboratory tests on cord blood demonstrated a red blood cell count of 163 x 10(4)/microL, and nucleated red cells numbering 1267 per 50 white cells in the live-birth case. Histologic examinations of fetal tissues demonstrated leukoerythroblastic reaction in the liver and spleen, granular hemosiderin deposition in hepatocytes and Kupffer cells, and bilirubin deposition in the intercellular space in the liver. This evidence indicates that, in some fetuses with intrauterine human parvovirus B-19 infection, hydropic changes may be induced by the sudden decrease in oxygen-carrying capacity of the blood due to severe anemia caused by the infection.

Infection with human parvovirus (B19), aplasia of the bone marrow and a rash in hereditary spherocytosis.

Infection with human parvovirus (HPV) B19 was diagnosed in persons with hereditary spherocytosis during an outbreak of erythema infectiosum. A 10-year-old boy had an aplastic crisis of the bone marrow and a maculopapular rash. His mother also had an aplastic crisis, fever, rash and a transiently acellular marrow. Another boy had an aplastic crisis, leucopenia and fever, but did not have a rash. Aplastic crisis and a rash have rarely before been observed together and attributed to infection with HPV.

Human parvovirus (B19) and erythema infectiosum.

An outbreak of erythema infectiosum ("fifth disease") was studied in Fukuoka, Japan, in 1980-1981. Human parvovirus (HPV) antigen was not detected in any patients, but anti-HPV, measured by countercurrent immunoelectrophoresis, was found in 33 of 34 affected children and in 21 (15%) of 141 children of the same ages without the disease. Immunoglobulin M class anti-HPV was present in all 25 children with erythema infectiosum tested. In a survey of hospital patients, the prevalence of anti-HPV detected by CIE was 12% in the cohort 5 to 9 years of age, 19% in the cohort 10 to 14 years, and 32 to 55% in the cohorts greater than or equal to 30 years. The antibody reactions in the cases of erythema infectiosum, which were already well established at the onset of disease, indicate that HPV was the cause of the outbreak.

Clinical observations on varicella-zoster vaccinees treated with immunosuppressants for a malignancy.

Between 1974 and 1983, 60 persons have been immunized at Kyushu University Hospital with a live attenuated varicella-zoster virus vaccine, Oka strain. The recipients were classified into 3 groups: those with a malignancy, those with the nephrotic syndrome and those with diseases not related to immuno-hematologic dyscrasia. The only adverse clinical reactions to the vaccine were skin rash with 3-30 vesicles and a body temperature of 38 C, which were seen in 2/21 (9.5%), 4/16 (25%) and 3/23 (13%) patients in the respective groups within 5 weeks after vaccination. From 6 months to 9 years after the vaccination, exogenous varicella infection occurred in 5/21 (23.8%), 1/16 (6.25%), and 0/23 (0%) patients in the respective groups. It is concluded that for patients with malignancies, revaccination is desirable to ensure the protective effect of the vaccine.