Development of recombinant diagnostic reagents based on pp85(U14) and p86(U11) proteins to detect the human immune response to human herpesvirus 7 infection.
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.
Stefan
A
A
Department of Experimental Pathology, Section of Microbiology and Virology, University of Bologna, Bologna, Italy.
De Lillo
M
M
Frascaroli
G
G
Secchiero
P
P
Neipel
F
F
Campadelli-Fiume
G
G
D000328:Adult; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D018604:Epitope Mapping; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000276:immunology; D016199:Herpesvirus 7, Human / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007231:Infant, Newborn; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D010455:Peptides; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins