Modulation of secretory functions in epithelia by adenovirus capsid proteins.
To evaluate the safety of adenovirus-derived capsid proteins for ocular gene delivery, we have investigated their effects on the morphology and function of the acinar epithelial cells of the lacrimal gland. These cells are responsible for basal and stimulated release of proteins and electrolytes into ocular fluid, a process essential in maintaining the health of the ocular surface. Acinar epithelial cells from rabbit lacrimal gland were exposed to one of two adenovirus serotype 5 capsid proteins, penton or knob (the carboxy-terminal fragment of the fiber capsid protein). Sustained (16-18 h) exposure to the penton at 20 microg/ml was associated with major changes in the organization of the regulated secretory pathway and cytoskeleton. These changes included an apparent loss of mature secretory vesicles enriched in rab3D around the apical lumen as well as a depletion of apical actin. The microtubule array in penton-treated acini also exhibited bundling and disorganization. None of these effects were elicited by exposure to knob protein. Penton treatment also caused a significant (p < or = 0.05) increase and decrease in basal and carbachol-stimulated release, respectively, of bulk protein. Competition studies showed that RGD peptide partially prevented the penton-induced changes in rab3D-enriched secretory vesicles and actin filaments. These findings suggest that the adenovirus penton protein compromises normal acinar secretory compartment organization and function and that these changes are due at least partly to penton-integrin interactions.
Hamm-Alvarez
Sarah F
SF
Department of Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, CA 90033, USA. shalvar@usc.edu
Xie
Jiansong
J
Wang
Yanru
Y
Medina-Kauwe
Lali K
LK
D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D016503:Drug Delivery Systems / Q000379:methods; D004353:Drug Evaluation, Preclinical; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D007765:Lacrimal Apparatus / Q000737:chemistry / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D010590:Phalloidine / Q000031:analogs & derivatives / Q000378:metabolism; D011817:Rabbits; D012235:Rhodamines / Q000494:pharmacology; D013997:Time Factors; D020692:rab3 GTP-Binding Proteins / Q000737:chemistry / Q000187:drug effects
D000199:Actins; D036022:Capsid Proteins; D039302:Integrin alphaVbeta3; D008869:Microtubule-Associated Proteins; D010446:Peptide Fragments; D012235:Rhodamines; C043322:penton protein, adenovirus; D010590:Phalloidine; D002217:Carbachol; C116711:Rab3d protein, mouse; D020692:rab3 GTP-Binding Proteins