Elmer W. Otteson

Regulation of Cellular Gene Expression in Endothelial Cells by Sin Nombre and Prospect Hill Viruses

Mechanisms of hantavirus-induced vascular leakage remain unknown. This study was initiated to determine whether hantavirus-induced changes in endothelial cell gene expression may provide insight into disease mechanisms. Additionally, by using pathogenic Sin Nombre virus (SNV) and non-pathogenic Prospect Hill virus (PHV), we wanted to identify cellular responses that are likely to differentiate pathogenic from nonpathogenic hantaviruses. Using the Affymetrix DNA Array, we found that PHV and SNV did not significantly differ in the number of activated genes (18 versus 14 genes) in infected endothelial cells at 4 h PI. However, a smaller group of genes (36) were up-regulated by PHV compared to SNV (175) at 12 h PI. Only two genes were down-regulated in SNV-infected cells. Expression of the functionally diverse group of genes was altered at an early stage of infection (4 and 12 h PI). The genes affected include putative anti-viral factors, transcription factors, growth factors, chemokines, receptors, structural proteins, metabolism, and kinases. Although many genes were activated in cells infected with SNV and PHV, overall cellular transcriptional responses were more altered by pathogenic SNV compared to non-pathogenic PHV.

Generation of a Recombinant Cytomegalovirus for Expression of a Hantavirus Glycoprotein

ABSTRACT A cytomegalovirus (CMV) was isolated from its natural host, Peromyscus maniculatus, and was designated Peromyscus CMV (PCMV). A recombinant PCMV was constructed that contained Sin Nombre virus glycoprotein G1 (SNV-G1) fused in frame to the enhanced green fluorescent protein (EGFP) gene inserted into a site homologous to the human CMV UL33 (P33) gene. The recombinant CMV was used for expression and immunization of deer mice against SNV-G1. The results of the study indicate that P. maniculatus could be infected with as few as 10 virus particles of recombinant virus. Challenge of P. maniculatus with either recombinant or wild-type PCMV produced no overt pathology in infected animals. P. maniculatus immunized with recombinant virus developed an antibody response to SNV and EGFP. When rechallenged with recombinant virus, animals exhibited an anamnestic response against SNV. Interestingly, a preexisting immune response against PCMV did not prevent reinfection with recombinant PCMV.

Prevalence of Hantavirus in Four Species of Neotoma from Arizona and Utah

Sin Nombre virus (SNV), a hantavirus, can cause severe respiratory illness and death in humans. The primary carrier is the deer mouse, Peromyscus maniculatus , but other species of rodents may be infected with the virus. We screened four species of woodrats ( Neotoma ) for hantavirus using both enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) tests. Woodrats were collected from a variety of habitats in Arizona and Utah. Only Neotoma lepida tested serologically positive in an ELISA assay for hantavirus and also contained SNV RNA. Moreover, across three distinct populations of N. lepida , individuals that tested serum positive were restricted to one population near Jericho, Utah. The prevalence of hantavirus in this population was 27% (4 of 15).

A possible role for nonsense suppression in the synthesis of a human cytomegalovirus 58-kDa virion protein

A 1.6-kb late mRNA originating from the HindIII R fragment of human cytomegalovirus (HCMV) encodes a 58-kDa virion phosphoprotein. Data presented support the hypothesis that this protein may be synthesized via the translational readthrough of an opal termination codon separating two open reading frames located in tandem. To our knowledge this is the first report of nonsense suppression as a means of regulating gene expression in HCMV.