scholarly journals Lack of an Immune Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term Doxycycline-Regulated Transgene Expression in Nonhuman Primates after Intramuscular Injection of Recombinant Adeno-Associated Virus

2002 ◽  
Vol 76 (22) ◽  
pp. 11605-11611 ◽  
Author(s):  
David Favre ◽  
Véronique Blouin ◽  
Nathalie Provost ◽  
Radec Spisek ◽  
Françoise Porrot ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Nonhuman Primates ◽  
Cynomolgus Macaque ◽  
Experimental Period ◽  
Adeno Associated Virus ◽  
Cellular Immune Responses ◽  
Transactivator Protein ◽  
Cellular Immune ◽  
Vector Dna

ABSTRACT We previously documented persistent regulation of erythropoietin (Epo) secretion in mice after a single intramuscular (i.m.) injection of a recombinant adeno-associated virus (rAAV) vector harboring both the tetracycline-dependent transactivator (rtTA) and the Epo cDNA (D. Bohl, A. Salvetti, P. Moullier, and J. M. Heard, Blood 92:1512-1517, 1998). Using the same vector harboring the cynomolgus macaque Epo cDNA instead, the present study evaluated the ability of the tetracycline-regulatable (tetR) system to establish long-term transgene regulation in nonhuman primates. The vector was administered i.m., after which 5-day induction pulses were performed monthly for up to 13 months by using doxycycline (DOX), a tetracycline analog. We show that initial inductions were successful in all individuals and that there was a tight regulation and a rapid deinduction pattern upon DOX withdrawal. For one macaque, regulation of Epo secretion was maintained during the entire experimental period; for the five remaining macaques, secreted Epo became indistinguishable from endogenous Epo upon repeated DOX inductions. We investigated the mechanism involved and showed that, except in the animal in which secretion persisted, delayed humoral and cellular immune responses were directed against the rtTA transactivator protein associated with the reduction of vector DNA in transduced muscles. This study provides some evidence that, when the immune system is not mobilized against the rtTA transactivator, the tetR-regulatable system is able to support long-term transgene regulation in the context of an rAAV in nonhuman primates. In addition, our results suggest potential improvements for vector design.

scholarly journals Adeno-Associated Viruses (AAV) and Host Immunity – A Race Between the Hare and the Hedgehog

2021 ◽  
Vol 12 ◽  
Author(s):  
Kleopatra Rapti ◽  
Dirk Grimm
Keyword(s):  
Immune Responses ◽  
Dorsal Root ◽  
Transgene Expression ◽  
State Of The Art ◽  
Host Immunity ◽  
Gene Therapies ◽  
Cellular Immune Responses ◽  
Cellular Immune

Adeno-associated viruses (AAV) have emerged as the lead vector in clinical trials and form the basis for several approved gene therapies for human diseases, mainly owing to their ability to sustain robust and long-term in vivo transgene expression, their amenability to genetic engineering of cargo and capsid, as well as their moderate toxicity and immunogenicity. Still, recent reports of fatalities in a clinical trial for a neuromuscular disease, although linked to an exceptionally high vector dose, have raised new caution about the safety of recombinant AAVs. Moreover, concerns linger about the presence of pre-existing anti-AAV antibodies in the human population, which precludes a significant percentage of patients from receiving, and benefitting from, AAV gene therapies. These concerns are exacerbated by observations of cellular immune responses and other adverse events, including detrimental off-target transgene expression in dorsal root ganglia. Here, we provide an update on our knowledge of the immunological and molecular race between AAV (the “hedgehog”) and its human host (the “hare”), together with a compendium of state-of-the-art technologies which provide an advantage to AAV and which, thus, promise safer and more broadly applicable AAV gene therapies in the future.

scholarly journals Five Years of Successful Inducible Transgene Expression Following Locoregional Adeno-Associated Virus Delivery in Nonhuman Primates with No Detectable Immunity

2019 ◽  
Vol 30 (7) ◽  
pp. 802-813 ◽  
Author(s):  
Mickaël Guilbaud ◽  
Marie Devaux ◽  
Celia Couzinié ◽  
Johanne Le Duff ◽  
Alice Toromanoff ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Nonhuman Primates ◽  
Adeno Associated Virus ◽  
Virus Delivery

scholarly journals Lack of Repeat Transduction by Recombinant Adeno-Associated Virus Type 5/5 Vectors in the Mouse Airway

2007 ◽  
Vol 81 (22) ◽  
pp. 12360-12367 ◽  
Author(s):  
Stephanie G. Sumner-Jones ◽  
Deborah R. Gill ◽  
Stephen C. Hyde
Keyword(s):  
Transgene Expression ◽  
Lung Diseases ◽  
Repeated Administration ◽  
Transduction Efficiency ◽  
Viral Gene ◽  
Rapid Rise ◽  
Adeno Associated Virus ◽  
Subsequent Failure

ABSTRACT While recombinant adeno-associated virus (rAAV) vectors promote long-term transgene expression in the lungs and other organs, the goal of correcting chronic inherited lung diseases such as cystic fibrosis with this type of viral gene transfer vector is limited by the requirement of achieving stable potent transgene expression, potentially requiring vector readministration. Here we evaluated the abilities of rAAV type 5/5 (rAAV5/5) vectors based on the genome and capsid of AAV5 to efficiently transduce the lungs and nasal epithelium of mice after repeated administration. Transduction efficiency as judged by reporter gene expression was markedly reduced on a second rAAV5/5 administration and effectively abolished on a third. Varying the period between administrations from 8 to 36 weeks did not allow efficient repeated administration. A rapid rise in anti-AAV5 antibodies was noted after rAAV5/5 vector administration that was sustained for the entire period of investigation (in some cases exceeding 9 months). Furthermore, this antibody response and subsequent failure to repeatedly administer the vector were not rescued by the in vivo expression of CTLA4Ig from an rAAV5/5 vector. These results suggest that without the development of an effective and clinically acceptable immunosuppression strategy, treatments for chronic diseases that require repeated administration of rAAV5/5 vectors will be unsuccessful.

scholarly journals Liver-Directed AAV8 Booster Vaccine Expressing Plasmodium falciparum Antigen Following Adenovirus Vaccine Priming Elicits Sterile Protection in a Murine Model

2021 ◽  
Vol 12 ◽  
Author(s):  
Mohammad Shahnaij ◽  
Mitsuhiro Iyori ◽  
Hiroaki Mizukami ◽  
Mayu Kajino ◽  
Iroha Yamagoshi ◽  
...  
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Malaria Vaccine ◽  
Infected Mosquito ◽  
Great Promise ◽  
Adeno Associated Virus ◽  
Boost Regimen ◽  
Cellular Immune Responses ◽  
Virus Serotype ◽  
Cellular Immune

Hepatocyte infection by malaria sporozoites is a bottleneck in the life-cycle of Plasmodium spp. including P. falciparum, which causes the most lethal form of malaria. Therefore, developing an effective vaccine capable of inducing the strong humoral and cellular immune responses necessary to block the pre-erythrocytic stage has potential to overcome the spatiotemporal hindrances pertaining to parasite biology and hepatic microanatomy. We recently showed that when combined with a human adenovirus type 5 (AdHu5)-priming vaccine, adeno-associated virus serotype 1 (AAV1) is a potent booster malaria vaccine vector capable of inducing strong and long-lasting protective immune responses in a rodent malaria model. Here, we evaluated the protective efficacy of a hepatotropic virus, adeno-associated virus serotype 8 (AAV8), as a booster vector because it can deliver a transgene potently and rapidly to the liver, the organ malaria sporozoites initially infect and multiply in following sporozoite injection by the bite of an infected mosquito. We first generated an AAV8-vectored vaccine expressing P. falciparum circumsporozoite protein (PfCSP). Intravenous (i.v.) administration of AAV8-PfCSP to mice initially primed with AdHu5-PfCSP resulted in a hepatocyte transduction rate ~2.5 times above that seen with intramuscular (i.m.) administration. This immunization regimen provided a better protection rate (100% sterile protection) than that of the i.m. AdHu5-prime/i.m. AAV8-boost regimen (60%, p < 0.05), i.m. AdHu5-prime/i.v. AAV1-boost (78%), or i.m. AdHu5-prime/i.m. AAV1-boost (80%) against challenge with transgenic PfCSP-expressing P. berghei sporozoites. Compared with the i.m. AdHu5-prime/i.v. AAV1-boost regimen, three other regimens induced higher levels of PfCSP-specific humoral immune responses. Importantly, a single i.v. dose of AAV8-PfCSP recruited CD8+ T cells, especially resident memory CD8+ T cells, in the liver. These data suggest that boost with i.v. AAV8-PfCSP can improve humoral and cellular immune responses in BALB/c mice. Therefore, this regimen holds great promise as a next-generation platform for the development of an effective malaria vaccine.

Effect of immunization with Ii-key modified HER2 (776-790) peptide vaccine (AE37) on immunologic responses in prostate cancer patients.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e16037-e16037
Author(s):  
Sonia A. Perez ◽  
Eleftheria Anastasopoulou ◽  
Efi Pappou ◽  
Panagiotis Tzonis ◽  
Stratos Bisias ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Patients ◽  
Peptide Vaccine ◽  
Treg Cells ◽  
Vaccine Responses ◽  
Cellular Immune Responses ◽  
Cellular Immune ◽  
Proliferation Assays

e16037 Background: We have shown that the AE37 vaccine (Ii-Key modified HER2(776-790) peptide) is safe and induces HER2/neu–specific cellular immune responses in patients with prostate cancer (Perez SA et al Clin. Cancer Res. 2010, 16:3495). We now present data from 4-year immunological assessments of prostate cancer patients who received AE37. Methods: Seventeen patients in a phase I study were given 6 doses of AE37 at monthly intervals and one additional dose a year after initiating treatment. Immunological testing to assess active versus suppressive immunity was conducted one month (intermediate-term immunomonitoring [ITI]) and 3 years (long-term immunomonitoring [LTI]) after the final dose of AE37. ELISPOT and proliferation assays were conducted to assess cytokine secretion and mitogenic response to antigen. DTH reactions were measured to assess in vivo immune response to antigen. All assays were conducted using native HER2(776-790) peptide (AE36). The percent Treg cells and ng/ml TGFβ were determined as markers for immune suppression. Results: Neither ELISPOT nor proliferation assays were statistically different at LTI compared to ITI. While clearly above pre-vaccine responses, the drop in DTH was statistically significant (p < 0.05). Similarly, the increase in Treg cells and circulating TGFβ was also statistically significant. An increase of >200 % in PSA-doubling time at any point during the study was observed in 6/17 patients, with 3 retaining this effect to 5 years. Conclusions: AE37 generates immunological memory associated with possible clinical efficacy in spite of Tregs and TGF-β levels returning at 4 years after being decreased for up to 6 months after initial AE37 vaccination. These results support further randomized testing of the AE37 vaccine. Clinical trial information: 2006-003299-37. [Table: see text]

Safety and Efficacy of an Adeno Associated Virus-Based Vector Carrying the Human Clotting Factor IX Gene (AAV5-hFIX) As Studied in Non-Human Primates

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4800-4800 ◽  
Author(s):  
Bart Nijmeijer ◽  
Harald Petry ◽  
Lisa Spronck ◽  
Corina Van Der Kruijssen ◽  
Jorgen Petersen ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Dose Group ◽  
Clinical Chemistry ◽  
Factor Ix ◽  
Clotting Factor ◽  
Adeno Associated Virus ◽  
Expression Levels ◽  
Normal Human ◽  
Dose Dependent ◽  
Vector Dna

Abstract Restoration of clotting Factor IX (FIX) expression through gene transfer is a promising option for the treatment of Hemophilia B. We have developed an Adeno-Associated Virus serotype 5-based vector containing the human coagulation factor IX gene (AAV5-hFIX), in our fully scalable GMP-compliant baculovirus-based production platform. To evaluate safety and efficacy of AAV5-hFIX in a relevant preclinical model, 4 groups of 3 cynomolgus macaques were dosed intravenously with 5x1011, 5x1012, 2.5x1013 and 0.93x1014 vector genome copies (gc) per kg body weight, respectively. Blood was sampled periodically, from 4 weeks pre-trial until necropsy at 26 weeks after dosing. Clinical chemistry and inflammation markers were assessed using standard techniques. Circulating human FIX protein levels were assessed by hFIX-specific ELISA. At necropsy, full histopathological examination was performed, and selected tissues were assessed for the presence of vector DNA and -RNA by quantitative PCR and RT-PCR, respectively. No signs of adverse reactions were observed in any of the animals throughout the in-life period. Clinical chemistry and inflammatory markers were unaffected by treatment with AAV5-hFIX. At necropsy, there were no macroscopic or microscopic tissue findings that could be attributed to the vector. Infusion of AAV5-hFIX resulted in dose-dependent circulating levels of hFIX protein. In the highest-dose group, hFIX levels peaked to 30% of normal human levels seven days after administration, and stabilized around 10-15% of normal human levels over the remaining observation period of 6 months. In the lowest-dose group, hFIX levels did not increase above 1%. Analysis of tissues after necropsy revealed dose-dependent vector DNA delivery to the liver, and concurrent dose-dependent transgene expression levels. No significant off-target expression was observed. In one animal, circulating hFIX levels decreased to baseline one month after vector administration. This decrease was shown to be caused by the development of xenoreactive hFIX-specific antibodies. This immune response was limited to sequestration of hFIX from the circulation, and did not entail T cell reactivity towards transduced hepatocytes as the transgene expression levels in the liver of this animal were comparable to those observed in its group mates. Based on the observed expression levels, and by extrapolating the dose one-on-one between non-human primates and humans, the Minimum Anticipated Biological Effect Level (MABEL) in man is 5x1012 gc/kg. In conclusion, administration of AAV5-hFIX to non-human primates was well tolerated without any noticeable adverse effects, and resulted in (i) dose-dependent transgene delivery to the liver, (ii) liver-specific transgene expression, and (iii) circulating human FIX protein to levels that are expected to be of significant clinical benefit in the setting of hemophilia B. Disclosures Nijmeijer: uniQure BV: Employment. Petry:uniQure B. V,: Employment. Spronck:uniQure BV: Employment. Van Der Kruijssen:uniQure BV: Employment. Petersen:uniQure: Consultancy. Salmon:uniQure B. V.: Employment.

scholarly journals Reduced schedule of human anti rabies immunization with Fuenzalida & Palacios vaccine

1989 ◽  
Vol 31 (1) ◽  
pp. 23-27 ◽  
Author(s):  
Carlos Roberto Zanetti ◽  
Silvana Regina Favoretto ◽  
Milene Silva Tino ◽  
Avelino Albas ◽  
Elizabeth Juliana G. Valentini ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Humoral Immune Response ◽  
The Other ◽  
Double Dose ◽  
Humoral Immune ◽  
Cellular Immune Responses ◽  
Lymphoblastic Transformation ◽  
Cellular Immune

The present study evaluates the humoral and cellular immune responses in 35 volunteers submited to short antirabies vaccination schedules with the Fuenzalida & Palacios vaccine based on the administration of doses on non consecutive days. The volunteers were divided into two groups. The first group received a total number of five doses given on days 0, 4, 7, 20 and 35. The other group received four doses, the first one being a double dose given on day 0 and than three other single doses on days 7, 20 and 35. The evaluation of humoral immune response was carried out by serum neutralization (SN) and indirect immunofluorescense (IIF) tests, while the cellular immune response was evaluated by lymphoblastic transformation assay (LTA) and skin test (ST). According to our results these reduced schedules elicited early and effective humoral and cellulafimmune responses to rabies antigen suggesting that new reduced schedules should be extensively studied in order to give the proper bases to the proposition of changes in the current long-term schedule.

Sign in / Sign up

Export Citation Format

Share Document