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Adenoviral capsid modulates secretory compartment organization and function in acinar epithelial cells from rabbit lacrimal gland

Gene Therapy volume 11, pages 970–981 (2004)Cite this article

Abstract

Although adenovirus (Ad) exhibits tropism for epithelial cells, little is known about the cellular effects of adenoviral binding and internalization on epithelial functions. Here, we examine its effects on the secretory acinar epithelial cells of the lacrimal gland, responsible for stimulated release of tear proteins into ocular fluid. Exposure of reconstituted rabbit lacrimal acini to replication-defective Ad for 16–18 h under conditions that resulted in >80% transduction efficiency did not alter cytoskeletal filament or biosynthetic/endosomal membrane compartment organization. Transduction specifically altered the organization of the stimulated secretory pathway, eliciting major dispersal of rab3D immunofluorescence from apical stores normally associated with mature secretory vesicles. Biochemical studies revealed that this dispersal was not associated with altered rab3D expression nor its release from cellular membranes. Ultraviolet (UV)-inactivated Ad elicited similar dispersal of rab3D immunofluorescence. In acini exposed to replication-defective or UV-inactivated Ad, carbachol-stimulated release of bulk protein and β-hexosaminidase were significantly (P⩽0.05) inhibited to an extent proportional to the loss of rab3D-enriched mature secretory vesicles associated with these treatments. We propose that the altered secretory compartment organization and function caused by Ad reflects changes in the normal maturation of secretory vesicles, and that these changes are caused by exposure to the Ad capsid.

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Acknowledgements

This work was supported by NIH Grants EY-13949 to SHA, EY12689 to MDT, P30 DK-48522 (Confocal Microscopy and Viral Vector Subcores, USC Center for Liver Diseases) and Doheny Eye Institute Core EY03040. Additional salary support to SHA was from NIH Grants EY-11386, EY-05081, NS-38246 and GM-59297. We thank Drs Austin Mircheff and Silvia da Costa for helpful discussions.

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  1. Y Wang and J Xie: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, USA

    Y Wang, J Xie, F A Yarber & S F Hamm-Alvarez

  2. Department of Physiology and Biophysics, University of Southern California, Los Angeles, CA, USA

    S F Hamm-Alvarez

  3. Department of Ophthalmology, University of Southern California, Los Angeles, CA, USA

    M D Trousdale & S F Hamm-Alvarez

  4. Institute for Genetic Medicine, University of Southern California, Los Angeles, CA, USA

    C Mazurek

  5. Gene Therapeutics Research Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    L K Medina-Kauwe

  6. Division of Digestive Diseases, University of California, Los Angeles, CA, USA

    N Kasahara

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Wang, Y., Xie, J., Yarber, F. et al. Adenoviral capsid modulates secretory compartment organization and function in acinar epithelial cells from rabbit lacrimal gland. Gene Ther 11, 970–981 (2004). https://doi.org/10.1038/sj.gt.3302247

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