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Open Access Highly Accessed Research article

Anti-West Nile virus activity of in vitro expanded human primary natural killer cells

Mingjie Zhang1*, Sylvester Daniel1, Yong Huang13, Caren Chancey1, Qingsheng Huang13, Ying F Lei14, Andriyan Grinev1, Howard Mostowski2, Maria Rios1 and Andrew Dayton1*

Author Affiliations

1 Laboratory of Molecular Virology, Center for Biologics Evaluation and Research, Food and Drug Administration, 1401 Rockville Pike, Rockville, MD 20892, USA

2 Cellular and Tissue Therapy Branch, Center for Biologics Evaluation and Research, Food and Drug Administration, 1401 Rockville Pike, Rockville, MD 20892, USA

3 Department of Life Science, Northwestern Polytechnical University, 127 Youyi Xilu, Xi'an, Shaanxi 710072, China

4 Department of Microbiology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China

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BMC Immunology 2010, 11:3  doi:10.1186/1471-2172-11-3

Published: 20 January 2010

Abstract

Background

Natural Killer (NK) cells are a crucial component of the host innate immune system with anti-viral and anti-cancer properties. However, the role of NK cells in West Nile virus (WNV) infection is controversial, with reported effects ranging from active suppression of virus to no effect at all. It was previously shown that K562-mb15-41BBL (K562D2) cells, which express IL-15 and 4-1BBL on the K562 cell surface, were able to expand and activate human primary NK cells of normal peripheral blood mononuclear cells (PBMC). The expanded NK cells were tested for their ability to inhibit WNV infection in vitro.

Results

Co-culture of PBMC with irradiated K562D2 cells expanded the NK cell number by 2-3 logs in 2-3 weeks, with more than 90% purity; upregulated NK cell surface activation receptors; downregulated inhibitory receptors; and boosted interferon gamma (IFN-γ) production by ~33 fold. The expanded NK (D2NK) cell has strong natural killing activity against both K562 and Vero cells, and killed the WNV infected Vero cells through antibody-dependent cellular cytotoxicity (ADCC). The D2NK cell culture supernatants inhibited both WNV replication and WNV induced cytopathic effect (CPE) in Vero cells when added before or after infection. Anti-IFN-γ neutralizing antibody blocked the NK supernatant-mediated anti-WNV effect, demonstrating a noncytolytic activity mediated through IFN-γ.

Conclusions

Co-culture of PBMC with K562D2 stimulatory cells is an efficient technique to prepare large quantities of pure and active NK cells, and these expanded NK cells inhibited WNV infection of Vero cells through both cytolytic and noncytolytic activities, which may imply a potential role of NK cells in combating WNV infection.