Stability of lentiviral vector-mediated transgene expression in the brain in the presence of systemic antivector immune responses.

Lentiviral vectors are promising tools for gene therapy in the CNS. It is therefore important to characterize their interactions with the immune system in the CNS. This work characterizes transgene expression and brain inflammation in the presence or absence of immune responses generated after systemic immunization with lentiviral vectors. We characterized transduction with SIN-LV vectors in the CNS. A dose-response curve using SIN-LV-GFP demonstrated detectable transgene expression in the striatum at a dose of 10(2), and maximum expression at 10(6), transducing units of lentiviral vector, with minimal increase in inflammatory markers between the lowest and highest dose of vector injected. Our studies demonstrate that injection of a lentiviral vector into the CNS did not cause a measurable inflammatory response. Systemic immunization after CNS injection, with the lentiviral vector expressing the same transgene as a vector injected into the CNS, caused a decrease in transgene expression in the CNS, concomitantly with an infiltration of inflammatory cells into the CNS parenchyma at the injection site. However, peripheral immunization with a lentiviral vector carrying a different transgene did not diminish transgene expression, or cause CNS inflammation. Systemic immunization preceding injection of lentiviral vectors into the CNS determined that preexisting antilentiviral immunity, regardless of the transgene, did not affect transgene expression. Furthermore, we showed that the transgene, but not the virion or vector components, is responsible for providing antigenic epitopes to the activated immune system, on systemic immunization with lentivirus. Low immunogenicity and prolonged transgene expression in the presence of preexisting lentiviral immunity are encouraging data for the future use of lentiviral vectors in CNS gene therapy. In summary, the lentiviral vectors tested induced undetectable activation of innate immune responses, and stimulation of adaptive immune responses against lentiviral vectors was effective in causing a decrease in transgene expression only if the immune response was directed against the transgene. A systemic immune response against vector components alone did not cause brain inflammation, possibly because vector-derived epitopes were not being presented in the CNS.

[1]  T. Kafri,et al.  Gene delivery by lentivirus vectors , 2007, Molecular biotechnology.

[2]  E. Chiocca,et al.  Oncolytic viruses: extreme treatment for an extreme disease. , 2006, Future microbiology.

[3]  J. Mallet,et al.  Lentiviral vectors with a defective integrase allow efficient and sustained transgene expression in vitro and in vivo , 2006, Proceedings of the National Academy of Sciences.

[4]  M. Caligiuri,et al.  Cyclophosphamide enhances glioma virotherapy by inhibiting innate immune responses , 2006, Proceedings of the National Academy of Sciences.

[5]  S. E. Barker,et al.  Effective gene therapy with nonintegrating lentiviral vectors , 2006, Nature Medicine.

[6]  N. Mazarakis,et al.  Lentivirus-mediated gene transfer to the central nervous system: therapeutic and research applications. , 2005, Human gene therapy.

[7]  P. Lowenstein,et al.  Neuronal expression of the transcription factor Gli1 using the Tα1 α-tubulin promoter is neuroprotective in an experimental model of Parkinson's disease , 2004, Gene Therapy.

[8]  N. Déglon,et al.  Lentiviral nigral delivery of GDNF does not prevent neurodegeneration in a genetic rat model of Parkinson's disease , 2004, Neurobiology of Disease.

[9]  R. Mandel,et al.  Circulating Anti-Wild-Type Adeno-Associated Virus Type 2 (AAV2) Antibodies Inhibit Recombinant AAV2 (rAAV2)-Mediated, but Not rAAV5-Mediated, Gene Transfer in the Brain , 2004, Journal of Virology.

[10]  P. Lowenstein.  Input virion proteins: cryptic targets of antivector immune responses in preimmunized subjects. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[11]  N. Déglon,et al.  Local GDNF expression mediated by lentiviral vector protects facial nerve motoneurons but not spinal motoneurons in SOD1G93A transgenic mice , 2004, Neurobiology of Disease.

[12]  P. Carmeliet,et al.  VEGF delivery with retrogradely transported lentivector prolongs survival in a mouse ALS model , 2004, Nature.

[13]  A. Annoni,et al.  Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice. , 2004, Blood.

[14]  L. Naldini,et al.  Correction of metachromatic leukodystrophy in the mouse model by transplantation of genetically modified hematopoietic stem cells. , 2004, The Journal of clinical investigation.

[15]  Theresa A. Storm,et al.  Rapid Uncoating of Vector Genomes Is the Key toEfficient Liver Transduction with Pseudotyped Adeno-Associated VirusVectors , 2004, Journal of Virology.

[16]  M. Hayden,et al.  Long-term lentiviral-mediated expression of ciliary neurotrophic factor in the striatum of Huntington's disease transgenic mice , 2004, Experimental Neurology.

[17]  N. Chirmule,et al.  Immunity to adenovirus and adeno-associated viral vectors: implications for gene therapy , 2003, Gene Therapy.

[18]  J. Bennett.  Immune response following intraocular delivery of recombinant viral vectors , 2003, Gene Therapy.

[19]  P. Lowenstein,et al.  Inflammation and adaptive immune responses to adenoviral vectors injected into the brain: peculiarities, mechanisms, and consequences , 2003, Gene Therapy.

[20]  J. Yewdell,et al.  Viral interference with antigen presentation , 2002, Nature Immunology.

[21]  P. Lowenstein,et al.  Adenovirus Binding to the Coxsackievirus and Adenovirus Receptor or Integrins Is Not Required To Elicit Brain Inflammation but Is Necessary To Transduce Specific Neural Cell Types , 2002, Journal of Virology.

[22]  L. Naldini,et al.  Efficient gene delivery and targeted expression to hepatocytes in vivo by improved lentiviral vectors. , 2002, Human gene therapy.

[23]  S. Kochanek,et al.  Preexisting antiadenoviral immunity is not a barrier to efficient and stable transduction of the brain, mediated by novel high-capacity adenovirus vectors. , 2001, Human gene therapy.

[24]  A. Consiglio,et al.  In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice , 2001, Nature Medicine.

[25]  M. Castro,et al.  Gene Transfer into Neural Cells In Vitro Using Adenoviral Vectors , 2000, Current protocols in neuroscience.

[26]  P. Lowenstein,et al.  Gene Transfer into Rat Brain Using Adenoviral Vectors , 2000, Current protocols in neuroscience.

[27]  J. Bloch,et al.  Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease. , 2000, Science.

[28]  S. Kochanek,et al.  Peripheral infection with adenovirus causes unexpected long-term brain inflammation in animals injected intracranially with first-generation, but not with high-capacity, adenovirus vectors: toward realistic long-term neurological gene therapy for chronic diseases. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  L. Ailles,et al.  Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences , 2000, Nature Genetics.

[30]  Charles R. Gerfen,et al.  Current Protocols In Neuroscience , 1999 .

[31]  L Naldini,et al.  Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector , 1997, Journal of virology.

[32]  F. Gage,et al.  In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector , 1996, Science.

[33]  D. C. Sterio.  The unbiased estimation of number and sizes of arbitrary particles using the disector , 1984, Journal of microscopy.

[34]  E. Chiocca,et al.  Personalizing oncolytic virotherapy? , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[35]  J. Uney,et al.  Lentiviral-mediated delivery of Bcl-2 or GDNF protects against excitotoxicity in the rat hippocampus. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[36]  N. Déglon,et al.  Lentiviral-mediated gene transfer to model triplet repeat disorders. , 2004, Methods in molecular biology.

[37]  P. Lowenstein.  Immunology of viral-vector-mediated gene transfer into the brain: an evolutionary and developmental perspective. , 2002, Trends in immunology.

[38]  Luigi Naldini,et al.  Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics , 2001, Nature Medicine.

[39]  P. Lowenstein,et al.  Acute direct adenoviral vector cytotoxicity and chronic, but not acute, inflammatory responses correlate with decreased vector-mediated transgene expression in the brain. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[40]  J. Yewdell,et al.  Mechanisms of exogenous antigen presentation by MHC class I molecules in vitro and in vivo: implications for generating CD8+ T cell responses to infectious agents, tumors, transplants, and vaccines. , 1999, Advances in immunology.