Immunological analysis of the tegument phosphoprotein ppUL83 of human cytomegalovirus.
Immunological properties of the tegument phosphoprotein, ppUL83, of human cytomegalovirus (HCMV), expressed using a replication deficient recombinant adenovirus vector (RAd83) are described. The initial characterisation of this protein was carried out by immunofluorescence (IF), immunoprecipitation (RIP) and immunoblotting using nine mouse monoclonal antibodies (Mabs) directed against five linear and four conformational epitopes of ppUL83. The reactivity of the recombinant protein with the Mabs was similar to that observed with native ppUL83, although, the kinetics of its expression was in agreement with expression derived from the HCMV major immediate early promoter (MIEP). The recombinant antigen was used successfully in an Enzyme Immunoassay (EIA) for the detection of IgG class antibodies in 171 sequential sera taken from 21 heart transplant recipients. Comparison of HCMV-infected and RAd83-infected cell extracts in this experiment showed that recombinant antigen could substitute whole virus extracts as a single well-characterised protein in EIA. Serum IgG avidity measurements, using the recombinant ppUL83, differentiated between primary and past HCMV infections in the population studied.
Zal
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Department of Cardiological Sciences, St George's Hospital Medical School, London, UK.
Pinto-Basto
J
J
Milne
R
R
Li
M S
MS
Jeeachee
M
M
Baboonian
C
C
D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D003587:Cytomegalovirus / Q000737:chemistry*; D003673:Defective Viruses / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D010750:Phosphoproteins / Q000032:analysis* / Q000276:immunology; D011233:Precipitin Tests; D014763:Viral Matrix Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa