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  • Original Article
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Downregulation of human CD46 by adenovirus serotype 35 vectors

Gene Therapy volume 14, pages 912–919 (2007)Cite this article

Abstract

Human CD46 (membrane cofactor protein), which serves as a receptor for a variety of pathogens, including strains of measles virus, human herpesvirus type 6 and Neisseria, is rapidly downregulated from the cell surface following infection by these pathogens. Here, we report that replication-incompetent adenovirus (Ad) serotype 35 (Ad35) vectors, which belong to subgroup B and recognize human CD46 as a receptor, downregulate CD46 following infection. A decline in the surface expression of CD46 in human peripheral blood mononuclear cells was detectable 6 h after infection, and reached maximum (72%) 12 h after infection. Ad35 vector-induced downregulation of surface CD46 levels gradually recovered after the removal of Ad35 vectors, however, complete recovery of CD46 expression was not observed even at 96 h after removal. The surface expression of CD46 was also reduced after incubation with fiber-substituted Ad serotype 5 (Ad5) vectors bearing Ad35 fiber proteins, ultraviolet-irradiated Ad35, vectors and recombinant Ad35 fiber knob proteins; in contrast, conventional Ad5 vectors did not induce surface CD46 downregulation, suggesting that the fiber knob protein of Ad35 plays a crucial role in the downregulation of surface CD46 density. These results have important implications for gene therapy using CD46-utilizing Ad vectors and for the pathogenesis of Ads that interact with CD46.

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Abbreviations

Ad:

adenovirus

APC:

allophycocyanin

BSA:

bovine serum albumin

CAR:

coxsackievirus and adenovirus receptor

FBS:

fetal bovine serum

FITC:

fluorescein isothiocyanate

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

GFP:

green fluorescence protein

HHV6:

herpesvirus type 6

MV:

measles virus

MVH:

measles virus hemagglutinin

MFI:

mean fluorescence intensity

MOI:

multiplicity of infection

PBMCs:

peripheral blood mononuclear cells

PBS:

phosphate-buffered saline

RT-PCR:

reverse transcriptase-polymerase chain reaction

SCRs:

short consensus repeats

VP:

vector particle

PE:

phycoerythrin.

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Acknowledgements

We thank Ms Naoko Funakoshi, Ms Tomomi Sasaki and Ms Noriko Tada for their technical assistance. We also thank Dr Tsukasa Seya (Hokkaido University, Japan) for providing anti-CD46 rabbit serum. This work was supported by grants for Health and Labour Sciences Research from the Ministry of Health, Labour, and Welfare of Japan, and by Grants-in-Aid for Scientific Research on Priority Areas (B).

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Authors and Affiliations

  1. Laboratory of Gene Transfer and Regulation, National Institute of Biomedical Innovation, Osaka, Japan

    F Sakurai, K Akitomo, K Kawabata & H Mizuguchi

  2. Pharmaceuticals and Medical Devices Agency, Tokyo, Japan

    T Hayakawa

  3. Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan

    H Mizuguchi

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  1. F Sakurai
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Correspondence to H Mizuguchi.

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Sakurai, F., Akitomo, K., Kawabata, K. et al. Downregulation of human CD46 by adenovirus serotype 35 vectors. Gene Ther 14, 912–919 (2007). https://doi.org/10.1038/sj.gt.3302946

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