Stunned silence: gene expression programs in human cells infected with monkeypox or vaccinia virus - PubMed (original) (raw)

Stunned silence: gene expression programs in human cells infected with monkeypox or vaccinia virus

Kathleen H Rubins et al. PLoS One. 2011.

Abstract

Poxviruses use an arsenal of molecular weapons to evade detection and disarm host immune responses. We used DNA microarrays to investigate the gene expression responses to infection by monkeypox virus (MPV), an emerging human pathogen, and Vaccinia virus (VAC), a widely used model and vaccine organism, in primary human macrophages, primary human fibroblasts and HeLa cells. Even as the overwhelmingly infected cells approached their demise, with extensive cytopathic changes, their gene expression programs appeared almost oblivious to poxvirus infection. Although killed (gamma-irradiated) MPV potently induced a transcriptional program characteristic of the interferon response, no such response was observed during infection with either live MPV or VAC. Moreover, while the gene expression response of infected cells to stimulation with ionomycin plus phorbol 12-myristate 13-acetate (PMA), or poly (I-C) was largely unimpaired by infection with MPV, a cluster of pro-inflammatory genes were a notable exception. Poly(I-C) induction of genes involved in alerting the innate immune system to the infectious threat, including TNF-alpha, IL-1 alpha and beta, CCL5 and IL-6, were suppressed by infection with live MPV. Thus, MPV selectively inhibits expression of genes with critical roles in cell-signaling pathways that activate innate immune responses, as part of its strategy for stealthy infection.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1

Figure 1. Host Gene Expression Overview.

An overview of the gene expression patterns of 5745 genes whose transcript levels changed, by a factor of at least 3, from their initial levels in at least one of the 134 samples taken from time courses viral infection or mock infection in macrophages from two human donors, human dermal fibroblasts, or HELA cells. In this display, genes were hierarchically clustered based on similarities in the patterns of variation of their transcript levels . In this display, each row represents a single gene, and each column represents a single cell sample from an infection or mock infection time course. The samples from each time course are ordered from left to right. Red and green colors represent expression levels greater or less, respectively, than baseline values (average of 2–5 samples taken immediately prior to infection, 0 hour time point). Grey indicates missing or excluded data. The intensity of the color reflects the magnitude of the change from baseline (Time zero), as indicated by the color scales at the bottom of each panel. Mock = mock infection (media only), V-WR = Vaccinia-Western Reserve, V-NY = Vaccinia-New York Board of Health (Wyeth, Dryvax), MPV = killed (gamma irradiated) Monkeypox-Zaire, MPV = Monkeypox-Zaire. Names of a few selected genes or categories of genes from clusters with distinctive patterns of induction or repression, are indicated to the right of enlarged views of the corresponding clusters, in panel B. Numbers to the right of panel A indicate the positions of the clusters shown enlarged and identified by the corresponding numbers in panel B. The vertical grey bars to the right of panel A indicate the positions of the genes encoding ribosomal proteins, mitochondrial proteins and proteosome components, shown in an enlarged view in panel B). For a comprehensive, browseable view of the gene expression programs depicted here, see the supporting information. The complete raw data from this experiment are available at:

www.smd.stanford.edu

or the GEO database (accession number GSE24125).

Figure 2

Figure 2. Stimulation of primary human fibroblasts.

Fibroblasts were treated for 24 hours with each of 6 different molecular stimuli, or with a mock treatment. 3754 genes whose expression level increased or decreased by at least a factor of three in response to one or more of the treatments are included in this display; each column represents the changes in abundance of transcripts from each of these genes after 24 hours of treatment with one of the agents. For each molecular stimulus, results from two replicate treatments are shown. The genes were hierarchically clustered, and their expression represented by a color scale as in Figure 1. The brightest red and green colors in this display correspond to increases or decreases in transcript abundance by factors of at least 32 relative to time zero. 0hr = pre-stimulation timepoint, all other timepoints taken 24 hours post stimulation; mock (PBS), IFNα = Interferon alpha, TNF-alpha = tumor necrosis factor alpha, I+P = ionomycin and phorbal 12-myristate 13-acetate, poly(I-C) = polyinosinic-polycytidylic acid, LPS = Escherichia coli 055:B5 lipopolysaccharide, Dex = Dexamethasone. Names of a few selected genes from clusters with distinctive patterns of induction by one or more of these agents are indicated to the right of the corresponding clusters, marked by the vertical black lines. For a comprehensive, browseable view of the transcript profiles characteristic of each response see the supporting information. The complete raw data from this experiment are available at:

www.smd.stanford.edu

or the GEO database (accession number GSE24125).

Figure 3

Figure 3. Effects of infection with MPV on the transcriptional responses of fibroblasts to poly-IC or PMA+ionomycin.

(A) Fibroblasts were infected with MPV at an MOI of 10, or mock-infected, 4 hours prior to treatment with either poly(I-C) or PMA+ionomycin. Cells were harvested and RNA samples analyzed at 12 and 24 hours after treatment of mock-infected cells (16 and 28 hours after mock-infection), at 1, 2, 6 and 12 hours after poly(I-C) treatment (5, 6, 10 and 16 hours after MPV infection), and at 1, 2, 6, 12 and 24 hours after PMA and ionomycin treatment (5, 6, 10 and 16 hours after MPV infection) of MPV-infected cells. The samples from each time course are ordered from left to right. For this display, genes were clustered and expression levels represented by a color scale as in Figures 1 and 2. The brightest red and green colors in this display correspond to increases or decreases in transcript abundance by factors of at least 32 relative to time zero. Mock = mock infection (media only), MPV = Monkeypox-Zaire, I+P = treated with ionomycin and PMA 4 hours after infection or mock-infection, Poly(I-C) = treated with poly(I-C) 4 hours after infection or mock-infection. Names of a few selected genes from 4 clusters, all induced by poly(I-C) in the mock-infected cells but not in the MPV-infected cells, are indicated to the right of the corresponding clusters, marked by the vertical black lines. For a comprehensive, browseable view of the gene expression programs depicted here, see the supporting information. The complete raw data from this experiment are available at:

www.smd.stanford.edu

or the GEO database (accession number GSE24125).

References

    1. Esposito JJ, Fenner F. Poxviruses. In: Fields BN, Knipe DM, Howley PM, Chanock RM, Melnick J, editors. Virology. 3rd ed. Philadelphia, , PA: Lippincott-Raven; 2001. pp. 2885–2921.
    1. Heymann DL, Szczeniowski M, Esteves K. Re-emergence of monkeypox in Africa: a review of the past six years. Br Med Bull. 1998;54:693–702. - PubMed
    1. Fenner F, Anderson DA, Arita I, Jezek Z, Ladnyi ID. Smallpox and its Eradication. Geneva: World Health Organization; 1988.
    1. Reed KD, Melski JW, Graham MB, Regnery RL, Sotir MJ, et al. The detection of monkeypox in humans in the Western Hemisphere. N Engl J Med. 2004;350:342–350. - PubMed
    1. Di Giulio DB, Eckburg PB. Human monkeypox: an emerging zoonosis. Lancet Infect Dis. 2004;4:15–25. - PMC - PubMed

Publication types

MeSH terms

Grants and funding

LinkOut - more resources