Induction of interferon-γ from natural killer cells by immunostimulatory CpG DNA is mediated through plasmacytoid-dendritic-cell-produced interferon-α and tumour necrosis factor-α

Jason Marshall

Darren Heeke

Christi Abbate

Priscilla Yee

Gary Van Nest

Dynavax Technologies Corporation, Berkeley, CA, USA

Correspondence: Dr Jason D. Marshall, Dynavax Technologies Corporation, 2929 7th Street, Suite 100, Berkeley, 94710 CA, USA. Email: moc.xavd@llahsramj Senior author: Gary Van Nest, email: moc.xavd@tsennavg

Dynavax Technologies Corporation, Berkeley, CA, USA

Received 2005 Mar 22; Revised 2005 Jul 26; Accepted 2005 Aug 26.

Abstract

Immunostimulatory sequences (ISS) that contain CpG motifs have been demonstrated to exert antipathogen and antitumour immunity in animal models through several mechanisms, including the activation of natural killer (NK) cells to secrete interferon-γ (IFN-γ) and to exert lytic activity. Since NK cells lack the ISS receptor TLR9, the exact pathway by which NK cells are activated by ISS is unclear. We determined that ISS-induced IFN-γ from NK cells is primarily dependent upon IFN-α release from plasmacytoid dendritic cells (PDCs), which directly activates the NK cell. However, further analysis indicated that other PDC-released soluble factor(s) may contribute to IFN-γ induction. Indeed, tumour necrosis factor-α (TNF-α) was identified as a significant contributor to ISS-mediated activation of NK cells and was observed to act in an additive fashion with IFN-α in the induction of IFN-γ from NK cells and to up-regulate CD69 expression on NK cells. This activity of TNF-α, however, was dependent upon the presence of PDC-derived factors such as type I interferon. These results illustrate an important function for type I interferon in innate immunity, which is not only to activate effectors like NK cells directly, but also to prime them for enhanced activation by other factors such as TNF-α.

Keywords: CpG DNA, interferon-α, interferon-γ, natural killer cells, plasmacytoid dendritic cells

Abstract

Human PDCs were stimulated with 5 μg/ml C274 or negative control CpG-C for 10 hr and then RNA was extracted and analysed via real-time PCR. The expression levels of 10 genes were normalized to ubiquitin expression. Data are presented as the mean of fold induction over medium control (given the value of 1·0) with SEM for four donors, and are representative of two experiments. Freshly isolated PDCs express mRNA for these genes at low levels similar to those from cells cultured for 10 hr in medium (used as the control).

Acknowledgments

We are grateful to O. Devergne (Paris University, Paris) for the generous gift of anti-IL-27, to C. Fressola (Dynavax), L. Tracy and N. Lynn (Advanced Bioscience Resources, Alameda, CA) for phlebotomy services, and to N. Khounchanh (Dynavax) for technical support.

Acknowledgments

Abbreviations

ISS	immunostimulatory sequences
ISS-PDC SN	SN derived from ISS-stimulated PDCs
PDC	plasmacytoid dendritic cell
SN	supernatant

Abbreviations

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