title: Recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream.
abstract: We have constructed a recombinant adeno-associated virus serotype 2 vector encoding human interleukin 10 (rAAVhIL10). IL-10 is a potent antiinflammatory/immune cytokine, which has received growing attention for its therapeutic potential. Human IL-10 (hIL-10) production was virus dose dependent after in vitro infection of HSG cells, a human submandibular gland cell line. The vector-derived hIL-10 produced was biologically active, as the medium from rAAVhIL10-infected HSG cells caused a dose-dependent blockade of IL-12 secretion from spleen cells of IL-10 knockout mice challenged with heat-killed Brucella abortus. Administration of rAAVhIL10 (10(10) genomes per gland) to both mouse submandibular glands led to hIL-10 secretion into the bloodstream (approximately 1-5 pg/ml), that is, in an endocrine manner, which was stable for approximately 2 months. Salivary gland administration of rAAVhIL10 under experimental conditions was more efficacious than intravenous administration (approximately 0.5-0.7 pg/ml). Also, hIL-10 was readily secreted in vitro from organ cultures of minced submandibular glands infected with rAAVhIL10, 6 or 8 weeks earlier. Consistent with these results, hIL-10 mRNA was detected by reverse transcription-polymerase chain reaction in submandibular glands of mice infected with rAAVhIL10 but not from control mice. At these doses, little to no hIL-10 was detected in mouse saliva. Using a rAAV serotype 2 vector encoding beta-galactosidase, we observed that the primary parenchymal target cells were ductal. These findings represent the first report of rAAV use to target exocrine glands for systemic secretion of a therapeutic protein, and support the notion that rAAV serotype 2 vectors may be useful in salivary glands for local (periglandular) and systemic gene-based protein therapeutics.
authors.affiliation: Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yamano
authors.forename: Li-Yun
authors.initials: LY
authors.lastname: Huang
authors.forename: Chuantian
authors.initials: C
authors.lastname: Ding
authors.forename: John A
authors.initials: JA
authors.lastname: Chiorini
authors.forename: Corinne M
authors.initials: CM
authors.lastname: Goldsmith
authors.forename: Robert B
authors.initials: RB
authors.lastname: Wellner
authors.forename: Basil
authors.initials: B
authors.lastname: Golding
authors.forename: Robert M
authors.initials: RM
authors.lastname: Kotin
authors.forename: Dorothy E
authors.initials: DE
authors.lastname: Scott
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Baum
mesh_terms: D000818:Animals; D001407:Bacillus; D019556:COS Cells; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D016753:Interleukin-10 / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D012333:RNA, Messenger; D011995:Recombination, Genetic; D012469:Salivary Glands / Q000166:cytology / Q000378:metabolism*; D013363:Submandibular Gland / Q000378:metabolism; D013623:Tail
chemical_list: D012333:RNA, Messenger; D016753:Interleukin-10