Polarized secretion of transgene products from salivary glands in vivo.
Previously (Kagami et al. Hum. Gene Ther. 1996;7:2177-2184) we have shown that salivary glands are able to secrete a transgene-encoded protein into serum as well as saliva. This result and other published data suggest that salivary glands may be a useful target site for vectors encoding therapeutic proteins for systemic delivery. The aim of the present study was to assess in vivo if transgene-encoded secretory proteins follow distinct, polarized sorting pathways as has been shown to occur "classically" in cell biological studies in vitro. Four first-generation, E1-, type 5 recombinant adenoviruses were used to deliver different transgenes to a rat submandibular cell line in vitro or to rat submandibular glands in vivo. Subsequently, the secretory distribution of the encoded proteins was determined. Luciferase, which has no signal peptide, served as a cell-associated, negative control and was used to correct for any nonspecific secretory protein release from cells. The three remaining transgene products tested, human tissue kallikrein (hK1), human growth hormone (hGH), and human alpha1-antitrypsin (halpha1AT), were predominantly secreted (>96%) in vitro. Most importantly, in vivo, after a parasympathomimetic secretory stimulus, both hK1 and hGH were secreted primarily in an exocrine manner into saliva. Conversely, halpha1AT was predominantly secreted into the bloodstream, i.e., in an endocrine manner. The aggregate results are consistent with the recognition of signals encoded within the transgenes that result in specific patterns of polarized protein secretion from rat submandibular gland cells in vivo.
Baum
B J
BJ
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, NIH, Bethesda, MD 20892, USA. bruce_j_baum@nih.gov
Berkman
M E
ME
Marmary
Y
Y
Goldsmith
C M
CM
Baccaglini
L
L
Wang
S
S
Wellner
R B
RB
Hoque
A T
AT
Atkinson
J C
JC
Yamagishi
H
H
Kagami
H
H
Parlow
A F
AF
Chao
J
J
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007610:Kallikreins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D021382:Protein Sorting Signals / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012469:Salivary Glands / Q000378:metabolism*; D019076:Transgenes; D000515:alpha 1-Antitrypsin / Q000235:genetics / Q000378:metabolism*
D021382:Protein Sorting Signals; D000515:alpha 1-Antitrypsin; D013006:Growth Hormone; D007610:Kallikreins