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Journal of Clinical Microbiology, December 1999, p. 3980-3985, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Development of Recombinant Diagnostic Reagents Based on pp85(U14) and p86(U11) Proteins To Detect the Human Immune Response to Human Herpesvirus 7 Infection

Alessandra Stefan,1 Margherita De Lillo,1 Giada Frascaroli,1 Paola Secchiero,2,3 Frank Neipel,4 and Gabriella Campadelli-Fiume1,*

Department of Experimental Pathology, Section of Microbiology and Virology, University of Bologna, Bologna,1 and Department of Embryology and Morphology, Section of Human Anatomy, University of Ferrara, Ferrara,3 Italy; Institute of Human Virology, University of Maryland at Baltimore, Baltimore, Maryland2; and Department of Clinic and Molecular Virology, University of Erlangen-Nurnberg, Erlangen, Germany4

Received 24 March 1999/Returned for modification 12 July 1999/Accepted 23 July 1999

Human antibodies raised in response to human herpesvirus 7 (HHV-7) infection are directed predominantly to one or more HHV-7-infected cell proteins with apparent molecular masses of about 85 to 89 kDa. The genes that encode these proteins are unknown. However, several HHH-7 genes that possibly encode proteins in this molecular mass range have been identified. Thus, the proteins encoded by open reading frame U14 (85 kDa) and U11 (86 kDa) were expressed as recombinant proteins in bacteria. Of 13 human serum specimens that recognized the 85- to 89-kDa protein(s) of HHV-7-infected cells by immunoblotting, 12 were also reactive with recombinant pp85(U14) and 8 were reactive with p86(U11). It is concluded that (i) the HHV-7 immunodominant protein is pp85(U14) and (ii) the lack of posttranslational modifications in procaryotically expressed pp85 does not adversely affect the reactivity of human sera. Monoclonal antibody (MAb) 5E1 is an HHV-7-specific MAb directed to pp85(U14). Here, the HHV-7-specific epitope in pp85(U14) was finely mapped to the C' terminal region between amino acid residues 484 and 502. However, as indicated by the low level of reactivity of human sera with the HHV-7-specific epitope recognized by MAb 5E1, human sera recognize additional epitopes of pp85(U14) that are required for their full reactivity.

* Corresponding author. Mailing address: Department of Experimental Pathology, Section of Microbiology and Virology, University of Bologna, Via San Giacomo 12, 40126 Bologna, Italy. Phone: 39 051 2094733 or 39 051 354734. Fax: 39 051 2094747. E-mail: campadel{at}kaiser.alma.unibo.it.

Journal of Clinical Microbiology, December 1999, p. 3980-3985, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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