Nuclear import as a barrier to infection of human umbilical vein endothelial cells by human cytomegalovirus strain AD169.
Human embryonal fibroblasts (HEF) are fully permissive for infection by human cytomegalovirus (HCMV) strain AD169, whereas human umbilical vein endothelial cells (HUVEC) seem to form an almost complete barrier to infection with this virus. To investigate this difference in permissiveness, HCMV infection of both cell types was studied using in situ hybridisation (ISH) as well as immunocytochemistry to detect viral DNA and viral proteins. At 2 h post-infection (p.i.), viral DNA was detected dispersed throughout the cytoplasm in both HEF and HUVEC, indicating that HCMV enters all cells of both cell types. At 4 h p.i., the viral DNA was found in the nucleus in HEF, and at the same time expression of immediate early (IE) antigen was found. In contrast, in HUVEC the expression of the IE proteins occurred in a limited number of cells at 8 h p.i., while in most HUVEC an accumulation of viral DNA around the nuclei was observed at this time point. In HUVEC, the nuclear localisation of viral DNA was detected 16 h p.i. in a minority of cells, indicating that transport of HCMV DNA into the nucleus is considerably slower in HUVEC than in HEF. Furthermore, the number of HUVEC containing HCMV DNA decreased about six-fold between 8 and 48 h p.i., indicating that HCMV DNA is either transported into the nucleus or eliminated. Apparently, the lower permissiveness of HUVEC for the HCMV strain AD169 relative to HEF is due to inefficient transport of HCMV DNA into the nuclei of infected HUVEC.
Department of Medical Microbiology, University of Maastricht, The Netherlands.
M E
ME
Slobbe-van Drunen
A T
AT
Hendrickx
R C
RC
Vossen
E J
EJ
Speel
M C
MC
van Dam-Mieras
C A
CA
Bruggeman
D000956:Antigens, Viral / Q000032:analysis; D002467:Cell Nucleus / Q000821:virology*; D002478:Cells, Cultured / Q000821:virology; D003587:Cytomegalovirus / Q000302:isolation & purification / Q000502:physiology*; D004279:DNA, Viral / Q000032:analysis; D004622:Embryo, Mammalian / Q000166:cytology; D004730:Endothelium, Vascular / Q000821:virology*; D005347:Fibroblasts / Q000821:virology; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology / Q000821:virology*; D014764:Viral Proteins / Q000032:analysis; D014779:Virus Replication
D000956:Antigens, Viral; D004279:DNA, Viral; D014764:Viral Proteins