Host response to the attenuated poxvirus vector NYVAC: upregulation of apoptotic genes and NF-kappaB-responsive genes in infected HeLa cells.
NYVAC has been engineered as a safe, attenuated vaccinia virus (VV) vector for use in vaccination against a broad spectrum of pathogens and tumors. Due to the interest in NYVAC-based vectors as vaccines and current phase I/II clinical trials with this vector, there is a need to analyze the human host response to NYVAC infection. Using high-density cDNA microarrays, we found 368 differentially regulated genes after NYVAC infection of HeLa cells. Clustering of the regulated genes identified six discrete gene clusters with altered expression patterns. Clusters 1 to 3 represented 47.5% of the regulated genes, with three patterns of gene activation kinetics, whereas clusters 4 to 6 showed distinct repression kinetics. Quantitative real-time reverse transcription-PCR analysis of selected genes validated the array data. Upregulated transcripts correlated with genes implicated in immune responses, including those encoding interleukin-1 receptor 2 (IL-1R2), IL-6, ISG-15, CD-80, and TNFSF7. NYVAC upregulated several intermediates of apoptotic cascades, including caspase-9, correlating with its ability to induce apoptosis. NYVAC infection also stimulated the expression of NF-kappaB1 and NF-kappaB2 as well as that of NF-kappaB target genes. Expression of the VV host range K1L gene during NYVAC infection prevented NF-kappaB activation, but not the induction of apoptosis. This study is the first overall analysis of the transcriptional response of human cells to NYVAC infection and provides a framework for future functional studies to evaluate this vector and its derivatives as human vaccines.
Guerra
Susana
S
Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología/CSIC, Ciudad Universitaria Cantoblanco, 28049 Madrid, Spain.
López-Fernández
Luis A
LA
Pascual-Montano
Alberto
A
Nájera
José Luis
JL
Zaballos
Angel
A
Esteban
Mariano
M
D051696:Activating Transcription Factors / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D040081:Protein Array Analysis; D015854:Up-Regulation; D014616:Vaccinia virus / Q000502:physiology*
D051696:Activating Transcription Factors; D001798:Blood Proteins; D016328:NF-kappa B; C506433:CASP9 protein, human; D053453:Caspase 9; D020169:Caspases