Identification and mapping of neutralizing epitopes of human parvovirus B19 by using human antibodies.
We identified and mapped the regions responsible for neutralization in the human parvovirus B19 structural protein by using region-specific human antibodies derived from seropositive blood donors. The region-specific antibodies were purified by using affinity columns coupled with synthetic peptides of the hydrophilic regions including the beta-turn structure deduced by the predicted secondary structure of VP2. Fifteen highly specific antibodies against the synthetic peptides were obtained. Ten of them were able to precipitate the radiolabeled virus. Six of them proved to be able to protect the colony-forming unit erythroid cells in human bone marrow cell cultures from injury by the virus. The sequences recognized by the six neutralizing antibodies were sites corresponding to amino acids 253 to 272, 309 to 330, 325 to 346, 359 to 382, 449 to 468, and 491 to 515 from the amino-terminal portion of VP2. These observations suggest that the neutralizing epitopes were distributed in the region from amino acid 253 in the amino-terminal portion of VP2 to the carboxyl terminus of VP2.
Sato
H
H
Clinical Laboratory, Kyushu University Hospital, Fukuoka, Japan.
Hirata
J
J
Kuroda
N
N
Shiraki
H
H
Maeda
Y
Y
Okochi
K
K
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010321:Parvoviridae / Q000276:immunology*; D010322:Parvoviridae Infections / Q000276:immunology*; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011487:Protein Conformation
D000914:Antibodies, Viral; D000939:Epitopes; D010455:Peptides