Presence of human cytomegalovirus (HCMV) immediate early mRNA but not ppUL83 (lower matrix protein pp65) mRNA in polymorphonuclear and mononuclear leukocytes during active HCMV infection.
During an active human cytomegalovirus (HCMV) infection, leukocytes, harbouring the HCMV lower matrix protein pp65 (ppUL83) are present in the peripheral blood and can be detected with the HCMV antigenaemia assay. In the present study, it was investigated whether the presence of pp65 in these cells was due to transcription of the virus genome or might be the result of uptake of this viral protein. Peripheral blood leukocytes of transplant recipients and AIDS patients with an active HCMV infection were investigated for the presence of HCMV immediate early (IE) antigen and pp65 using well characterized monoclonal antibodies, and for the presence of the corresponding mRNAs using non-radioactive in situ hybridization. Both mononuclear and polymorphonuclear cells were found to contain IE antigen and pp65. However, only mRNAs encoding IE antigen were found in these cells, whereas mRNAs encoding pp65 were not detected. In contrast, both IE antigen and pp65, as well as their corresponding mRNAs, were detected in the circulating late-stage HCMV-infected endothelial cells that were also present in the leukocyte fractions. These findings demonstrate that a restricted viral gene expression (transcription of IE genes) does occur in mononuclear and polymorphonuclear leukocytes. However, the abundant presence of the early antigen pp65 without detectable presence of the corresponding mRNA in these cells strongly indicates uptake of this protein by the phagocytic leukocytes, rather than de novo synthesis.
Grefte
A
A
Department of Clinical Immunology, University of Groningen, The Netherlands.
Harmsen
M C
MC
van der Giessen
M
M
Knollema
S
S
van Son
W J
WJ
The
T H
TH
D000956:Antigens, Viral / Q000032:analysis* / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000032:analysis* / Q000235:genetics; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods; D009000:Monocytes; D009504:Neutrophils; D010586:Phagocytes; D010587:Phagocytosis; D010750:Phosphoproteins / Q000032:analysis* / Q000235:genetics; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis*; D014158:Transcription, Genetic; D014763:Viral Matrix Proteins / Q000032:analysis* / Q000235:genetics
D000956:Antigens, Viral; D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D015347:RNA Probes; D012333:RNA, Messenger; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; C044582:immediate-early proteins, cytomegalovirus