Adenoviral-mediated Gene Transfer to Mouse Salivary Glands

Abstract

Adenoviral vectors effectively transfer genes to rat salivary glands. However, potent immune responses limit their use in vivo. Mice offer more opportunities than rats for the study of these immune processes. We first established conditions for infection of mouse salivary glands, with an adenoviral vector. The effects of time, viral dose, viral diluent buffer volume, and dexamethasone on expression of a transgene, luciferase, were determined by means of the recombinant vector AdCMVluc. Optimal luciferase expression was observed when the vector was suspended in 50 microL of buffer. This volume completely filled the gland parenchyma and slightly distended the capsule. Dexamethasone increased immediate transgene expression and reduced the acute inflammation one day following viral administration, but did not alter subsequent mononuclear inflammation or transgene expression 14 or 28 days later. An adenoviral vector encoding either anti-inflammatory cytokine IL-4 or IL-10 was co-administered with AdCMVluc to increase transgene expression at 14 and 28 days. While this strategy did not extend the duration of luciferase expression, co-administration of AdCMVIL-10 with AdCMVluc almost completely eliminated the chronic inflammatory infiltrate in the glands after 28 days. This study demonstrates that adenoviral-mediated gene transfer to mouse submandibular glands is possible by intraductal cannulation and that reduction of either the acute or chronic inflammatory infiltrates was insufficient to increase long-term transgene expression in this tissue.

Ultrasound-assisted non-viral gene transfer to the salivary glands

“…The only clinical trial to date involving gene transfer Ultrasound-assisted gene transfer to the salivary glands MJ Passineau et al to the salivary glands has used an Ad vector, which Baum and colleagues have shown to elicit substantial inflammation in the gland, 23,29 presumably leading to functional impairment (which has been proposed to be mainly reversible 30 ). Our findings are in general agreement with earlier work in this field, indicating that transient physical disruption of cell membranes by local ultrasound-induced cavitation is not permanently injurious to the target cells.…”

Enhanced transduction of mouse salivary glands with AAV5-based vectors

“…8,22,41 Briefly, male mice (7 weeks old) were anesthetized with ketamine (60 mg/ml, 1 ml/g body weight; Phoenix Scientific, St Joseph, MO, USA) and xylazine (8 mg/ml; Phoenix Scientific), by i.m. injection, then administered atropine (0.5 mg/kg body weight, i.m.…”

Salivary Glands: A Powerful Experimental System to Study Cell Biology in Live Animals by Intravital Microscopy

“…Notably, for viral-mediated gene therapy in humans, the SGs offer several advantages with respect to other organs: i) the encapsulation of the SG tissue prevents the dissemination of the virus in the rest of the body (Baum et al, 2004;Voutetakis et al, 2004), ii) in case of potential health issues the SGs can be removed since they are not essential for life, iii) the differentiation of the cells provides a relatively stable cell populations for non-integrating vectors, and iv) duacrine (both exocrine and endocrine) protein secretion allows to direct the expressed molecules into either the saliva or the blood stream (Baum et al, 2004). Numerous studies have shown successful gene transfer into both rat and mouse submandibular glands using viral-based approaches, which offer the advantage of a more robust expression of the transgenes (Andresen et al, 2009;Baum and Tran, 2006;Delporte et al, 1996;Honigman et al, 2001;Mastrangeli et al, 1994;Morita et al, 2011;Palaniyandi et al, 2011;Perez et al, 2011;Samuni et al, 2008;Wang et al, 2000;Zheng et al, 2009). However, non viral-mediated approaches have also been utilized, although limited to a small percentage of the cells in the parenchyma (Goldfine et al, 1997;Honigman et al, 2001;Niedzinski et al, 2003a;Niedzinski et al, 2003b;Passineau et al, 2010;Sramkova et al, 2009).…”