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Journal of Virology, October 2009, p. 10548-10556, Vol. 83, No. 20
0022-538X/09/$08.00+0 doi:10.1128/JVI.01250-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Baculovirus Transduction of Mesenchymal Stem Cells Triggers the Toll-Like Receptor 3 Pathway 
,
Guan-Yu Chen,1,
Hsiao-Chiao Shiah,1,
Hung-Ju Su,2
Chi-Yuan Chen,1
Yung-Jen Chuang,3
Wen-Hsin Lo,1
Jie-Len Huang,1
Ching-Kuang Chuang,1
Shiaw-Min Hwang,4* and
Yu-Chen Hu1*
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan,1 HTP Transcriptomics and Proteomics Department, Molecular Biomedical Technology Division, Biomedical Engineering Research Lab, Industrial Technology Research Institute, Hsinchu 310, Taiwan,2 Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan,3 Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 300, Taiwan4
Received 16 June 2009/ Accepted 26 July 2009
Human mesenchymal stem cells (hMSCs) can be genetically modified with viral vectors and hold promise as a cell source for regenerative medicine, yet how hMSCs respond to viral vector transduction remains poorly understood, leaving the safety concerns unaddressed. Here, we explored the responses of hMSCs against an emerging DNA viral vector, baculovirus (BV), and discovered that BV transduction perturbed the transcription of 816 genes associated with five signaling pathways. Surprisingly, Toll-like receptor-3 (TLR3), a receptor that generally recognizes double-stranded RNA, was apparently upregulated by BV transduction, as confirmed by microarray, PCR array, flow cytometry, and confocal microscopy. Cytokine array data showed that BV transduction triggered robust secretion of interleukin-6 (IL-6) and IL-8 but not of other inflammatory cytokines and beta interferon (IFN-β). BV transduction activated the signaling molecules (e.g., Toll/interleukin-1 receptor domain-containing adaptor-inducing IFN-β, NF-
B, and IFN regulatory factor 3) downstream of TLR3, while silencing the TLR3 gene with small interfering RNA considerably abolished cytokine expression and promoted cell migration. These data demonstrate, for the first time, that a DNA viral vector can activate the TLR3 pathway in hMSCs and lead to a cytokine expression profile distinct from that in immune cells. These findings underscore the importance of evaluating whether the TLR3 signaling cascade plays roles in the immune response provoked by other DNA vectors (e.g., adenovirus). Nonetheless, BV transduction barely disturbed surface marker expression and induced only transient and mild cytokine responses, thereby easing the safety concerns of using BV for hMSCs engineering.
* Corresponding author. Mailing address for S.-M. Hwang: Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 300, Taiwan. Phone: 886 3 522 3191, ext. 576. Fax: 886 3 521 4016. E-mail: hsm{at}firdi.org.tw. Mailing address for Y.-C. Hu: Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan. Phone: 886 3 571 8245. Fax: 886 3 571 5408. E-mail: yuchen{at}che.nthu.edu.tw
Published ahead of print on 5 August 2009.
Supplemental material for this article may be found at http://jvi.asm.org/.
G.-Y.C. and H.-C.S. contributed equally to this work.
Journal of Virology, October 2009, p. 10548-10556, Vol. 83, No. 20
0022-538X/09/$08.00+0 doi:10.1128/JVI.01250-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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