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Published on: November 1, 2011
Characterization of Parvovirus B19 genotype 2 in KU812Ep6 cells
Johannes Blümel1, Anna Maria Eis-Hübinger, Albert Stühler
1Paul-Ehrlich-Institut, Langen, Germany. blujo@pei.de
Abstract:
An infectious parvovirus B19 (B19V) genotype 2 variant was identified as a high-titer contaminant in a human plasma donation. Genome analysis revealed a 138-bp insertion within the p6 promoter. The inserted sequence was represented by an additional 30 bp from the end of the inverted terminal repeat adjacent to a 108-bp element found also, in inverted orientation, at the extreme right end of the unique sequence of the genome. However, despite the profound variations in the promoter region, the pattern of gene expression and DNA replication did not differ between genotype 1 and genotype 2 in permissive erythroid KU812Ep6 cells. Capsid proteins of both genotypes differ in their amino acid sequences. However, equivalent kinetics of virus inactivation at 56 degrees C or pH 4 indicated a comparable physicochemical stability of virus capsids. Sera from six individuals infected by B19V genotype 1 were investigated on cross-neutralization of B19V genotype 2 in vitro. Similar neutralization of both B19V genotypes was observed in sera from three individuals, while the sera from three other individuals showed weaker cross-neutralization for genotype 2. In conclusion, the in vitro replication characteristics and physical stability of B19V capsids are very similar between human parvovirus B19 genotypes 1 and 2, and cross-neutralization indicates a close antigenic relation of genotypes 1 and 2.
Insights
A parvovirus B19 (B19V) genotype 2 variant, found in human plasma, showed significant genomic changes but similar replication and stability to genotype 1. Cross-neutralization suggests a close antigenic relationship between B19V genotypes.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- Human parvovirus B19 (B19V) is a significant human pathogen.
- B19V exhibits genetic diversity, with distinct genotypes identified.
- Understanding genotype-specific characteristics is crucial for diagnostics and therapeutics.
Purpose of the Study:
- To characterize a novel B19V genotype 2 variant.
- To compare the biological and physicochemical properties of B19V genotypes 1 and 2.
- To assess the cross-neutralization potential between B19V genotypes.
Main Methods:
- Genome sequencing and analysis of a B19V genotype 2 variant.
- In vitro studies using permissive erythroid KU812Ep6 cells to assess gene expression and DNA replication.
- Physicochemical stability assays (heat and pH inactivation).
- In vitro cross-neutralization assays using human sera from B19V genotype 1-infected individuals.
Main Results:
- A B19V genotype 2 variant with a 138-bp insertion in the p6 promoter was identified as a plasma contaminant.
- Despite genomic variations, genotype 1 and 2 showed similar gene expression and DNA replication kinetics in permissive cells.
- Virus capsid proteins differed in amino acid sequences, but physicochemical stability (heat and pH inactivation) was comparable.
- Sera from B19V genotype 1-infected individuals exhibited variable cross-neutralization against genotype 2, with some showing similar and others weaker neutralization.
Conclusions:
- Human parvovirus B19 genotypes 1 and 2 share similar in vitro replication characteristics and capsid physicochemical stability.
- The genomic variations in the identified genotype 2 variant did not significantly alter its replication or stability.
- Cross-neutralization data indicate a close antigenic relationship between B19V genotypes 1 and 2, suggesting potential for cross-protection or cross-reactivity in diagnostic assays.
