Slow AAV2 clearance from the brain of nonhuman primates and anti-capsid immune response.
Adeno-associated virus serotype 2 (AAV2) has previously been reported to be a slowly uncoating virus in peripheral tissues, but persistence of intact vector in primate brain has not been explored. Because some neurological gene therapies may require re-administration of the same vector to patients, it seems important to understand the optimal timeframe in which to consider such repeat intervention. Surprisingly, convection-enhanced delivery of AAV2 into the thalamus of nonhuman primates (NHPs) resulted in robust staining of neurons with A20 antibody that detected intact AAV2 particles at ∼1.5 months after infusion. However, by 2.5 months, no A20 staining was visible. These data confirmed earlier findings of persistence of intact AAV2 particles in ocular and hepatic tissues. In order to probe the potential consequences of this persistence, we infused AAV2-human aromatic L-amino acid decarboxylase into left and right thalamus of three NHPs, with a 3-month delay between infusions. During that interval, we immunized each animal subcutaneously with AAV2 virus-like particles (empty vector) in order to induce strong anti-capsid humoral immunity. Various high neutralizing antibody titers were achieved. The lowest titer animal showed infiltration of B lymphocytes and CD8(+) T cells into both the secondary and primary infusion sites. In the other two animals, extremely high titers resulted in no transduction of the second site and, therefore, no lymphocytic infiltration. However, such infiltration was prominent at the primary infusion site in each animal and was associated with overt neuronal loss and inflammation.
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
L
http://orcid.org/0000-0001-8151-473X
L
Samaranch
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
P
P
Hadaczek
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
A P
AP
Kells
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
J R
JR
Bringas
Valley Biosystems, West Sacramento, CA, USA.
D
D
Stockinger
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
W
W
San Sebastian
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
M
M
Macayan
Valley Biosystems, West Sacramento, CA, USA.
S
S
Samineni
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
P
P
Pivirotto
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
J
J
Forsayeth
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
K S
KS
Bankiewicz
D000818:Animals; D001921:Brain / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002213:Capsid / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D000229:Dependovirus / Q000276:immunology / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D008253:Macaca mulatta; D008297:Male; D011323:Primates; D014161:Transduction, Genetic
D036022:Capsid Proteins