Host response to the attenuated poxvirus vector NYVAC: upregulation of apoptotic genes and NF-kappaB-responsive genes in infected HeLa cells.

Abstract

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.

• Authors: Susana Guerra, Luis A López-Fernández, Alberto Pascual-Montano, José Luis Nájera, Angel Zaballos, Mariano Esteban
• Journal: Journal of virology (J Virol) Vol. 80 Issue 2 Pg. 985-98 (Jan 2006) ISSN: 0022-538X [Print] United States
• PMID: 16379000 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Activating Transcription Factors
• Blood Proteins
• NF-kappa B
• CASP9 protein, human
• Caspase 9
• Caspases

Topics

• Activating Transcription Factors (genetics, metabolism)
• Apoptosis (genetics)
• Blood Proteins (genetics, metabolism)
• Caspase 9
• Caspases (genetics, metabolism)
• Gene Expression Profiling
• HeLa Cells (metabolism)
• Humans
• NF-kappa B (genetics, metabolism)
• Protein Array Analysis
• Up-Regulation
• Vaccinia virus (physiology)