Efficient intrathymic gene transfer following in situ administration of a rAAV serotype 8 vector in mice and nonhuman primates.
The thymus is the primary site of T-cell development and plays a key role in the induction of self-tolerance. We previously showed that the intrathymic (i.t.) injection of a transgene-expressing lentiviral vector (LV) in mice can result in the correction of a T cell-specific genetic defect. Nevertheless, the efficiency of thymocyte transduction did not exceed 0.1-0.3% and we were unable to detect any thymus transduction in macaques. As such, we initiated studies to assess the capacity of recombinant adeno-associated virus (rAAV) vectors to transduce murine and primate thymic cells. In vivo administration of AAV serotype 2-derived single-stranded AAV (ssAAV) and self-complementary AAV (scAAV) vectors pseudotyped with capsid proteins of serotypes 1, 2, 4, 5, and 8 demonstrated that murine thymus transduction was significantly enhanced by scAAV2/8. Transgene expression was detected in 5% of thymocytes and, notably, transduced cells represented 1% of peripheral T lymphocytes. Moreover, i.t. administration of scAAV2/8 particles in macaques, by endoscopic-mediated guidance, resulted in significant gene transfer. Thus, in healthy animals, where thymic gene transfer does not provide a selective advantage, scAAV2/8 is a unique tool promoting the in situ transduction of thymocytes with the subsequent export of gene-modified lymphocytes to the periphery.
Moreau
Aurélie
A
Institut National de la Santé et de la Recherche Médicale U649-Laboratoire de Thérapie Génique, CHU Hôtel-Dieu, Nantes, France.
Vicente
Rita
R
Dubreil
Laurence
L
Adjali
Oumeya
O
Podevin
Guillaume
G
Jacquet
Chantal
C
Deschamps
Jack Yves
JY
Klatzmann
David
D
Cherel
Yan
Y
Taylor
Naomi
N
Moullier
Philippe
P
Zimmermann
Valérie S
VS
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics* / Q000494:pharmacology; D016679:Genome, Viral / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008252:Macaca fascicularis / Q000378:metabolism*; D051379:Mice; D010641:Phenotype; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism*; D019076:Transgenes / Q000235:genetics*