Prokaryotic expression of the major capsid protein of human cytomegalovirus and antigenic cross-reactions with herpes simplex virus type 1.
The major capsid protein (MCP) of human cytomegalovirus (HCMV) was expressed in three portions as beta-galactosidase fusion proteins, covering about 75% of the open reading frame (ORF). Fusion protein SH 1 contained nucleotides 101 to 1243 of the ORF, fusion protein FS 1 contained nucleotides 1944 to 3089 and fusion protein SS 1 covered nucleotides 2624 to 3793. The recombinant proteins were tested for their immunoreactivity with human sera. Fusion protein FS 1 was found to represent the immunodominant region. The recombinant proteins were used to generate polyvalent rabbit antisera to investigate cross-reactivities with the major capsid protein (VP5) of herpes simplex virus type 1 (HSV-1). A monospecific antiserum raised against the fusion protein close to the N terminus of the MCP, as well as a monoclonal antibody and a monospecific rabbit antiserum directed against the viral MCP, cross-reacted with the VP5 as shown by immunoblotting and immunofluorescence. In order to detect common epitopes of the major capsid proteins of HCMV and HSV-1, the recombinant proteins were conjugated to CNBr-activated Sepharose and taken for purification of MCP antibodies from HCMV and HSV-1 seropositive individuals. Using this affinity chromatography method, cross-reactivity could be observed with HCMV- and HSV-positive human antisera in immunoblot experiments.
Rudolph
S A
SA
Institut für Klinische und Molekulare Virologie, Erlangen, F.R.G.
Kühn
J E
JE
Korn
K
K
Braun
R W
RW
Jahn
G
G
D000911:Antibodies, Monoclonal / Q000276:immunology; D002213:Capsid / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003429:Cross Reactions; D003587:Cytomegalovirus / Q000235:genetics* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D005822:Genetic Vectors; D006801:Humans; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification; D015151:Immunoblotting; D011993:Recombinant Fusion Proteins / Q000276:immunology; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics* / Q000276:immunology; D012867:Skin; D001616:beta-Galactosidase / Q000235:genetics
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase