Delivery of AAV5 Vectors to Salivary Glands

The construction of rAAV5 encoding green fluorescent protein, luciferase, or bone morphogenetic protein 6 (BMP-6) vector (AAV5-GFP, AAV5-Luc, or AAV5-Bmp6, respectively), has been described previously (9 (link),10 (link)). Vectors were delivered into the submandibular gland by retrograde instillation and into the lacrimal gland by injection as previously described (10 (link),11 (link)). Briefly, AAV5-GFP or AAV5-Bmp6 (10¹¹ particles/mouse [5 × 10¹⁰ particles/gland] in 100 μl) was delivered into the submandibular gland of 6–8-week-old C57BL/6J mice. A lower dose of AAV5-Luc (10⁹ particles/ gland) was coadministered to enable monitoring of the expression of the AAV5 vectors by Xenogeny live imaging of the animals (10 (link)) (see Supplementary Figure 1, on the Arthritis & Rheumatism web site at http://onlinelibrary.wiley.com/doi/10.1002/art.38123/abstract). To transduce the lacrimal gland, AAV5-Bmp6 or AAV5-Luc vector was injected at 10⁹ particles/gland/mouse directly into the lacrimal gland of 6–8-week-old C57BL/6J mice (10 (link)). The vector dose was chosen based on previously published results, which showed detectable transgene activity with administration of >10⁹ particles/ gland (12 (link),13 (link)).