title: Transduction, tropism, and biodistribution of AAV vectors in the lacrimal gland.
abstract: OBJECTIVE
  The lacrimal gland (LG) delivers defensive and metabolic factors to the ocular surface. These functions may be disrupted in several diseases, and for most of them there is no cure. The aim of this study is to investigate conditions and limitations for using adeno-associated virus (AAV) vectors as gene transfer agents to LG.
  
  
  METHODS
  Eight-week-old Balb/c mice were used to investigate route, gene expression, and time course of AAV gene vector transfer to LG. AAV vectors encoding firefly luciferase were administered to the LG and luciferase expression was evaluated in vivo by immunohistochemistry. Ocular surface and neutralizing antibodies were also evaluated.
  
  
  RESULTS
  The present work revealed that AAV vectors are able to delivery DNA to the LGs of mice. Direct injection had the highest level of transduction, and topical ocular drops the lowest. Overall, the AAV strain with highest transduction activity as measured by both luminescence and immunohistochemistry was AAV9, followed by AAV 5w8 and AAV5. Transduction was not different between sexes, could be detected as soon as 24 hours after injection, and lasted for at least 30 days (study termination). No tissue damage was observed when compared with controls. All vectors with detectable LG transduction induced neutralizing antibodies.
  
  
  CONCLUSIONS
  LG gene delivery by AAV vectors appears to be both safe and well tolerated. The choice of vector influences both the overall transduction activity, as well as the spread of vector to other organs. This work supports the use of AAV-mediated gene therapy for dry eye.
authors: Molecular Physiology and Therapeutic Branch, NIDCR, NIH, Bethesda, Maryland, USA. emrocha@fmrp.usp.br
authors: Eduardo M
authors: EM
authors: Rocha
authors: Giovanni
authors: G
authors: Di Pasquale
authors: Paola Perez
authors: PP
authors: Riveros
authors: Kathrina
authors: K
authors: Quinn
authors: Beverly
authors: B
authors: Handelman
authors: John A
authors: JA
authors: Chiorini
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000229:Dependovirus / Q000502:physiology*; D005260:Female; D005822:Genetic Vectors; D007124:Immunoenzyme Techniques; D007765:Lacrimal Apparatus / Q000378:metabolism* / Q000821:virology; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D014018:Tissue Distribution; D014161:Transduction, Genetic; D014162:Transfection; D056189:Viral Tropism / Q000235:genetics*
chemical_list: D057134:Antibodies, Neutralizing; D008156:Luciferases