The Experts below are selected from a list of 4098 Experts worldwide ranked by ideXlab platform
Larry C. Ewalt - One of the best experts on this subject based on the ideXlab platform.
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one time Gene Gun or intramuscular rabies dna vaccination of non human primates comparison of neutralizing antibody responses and protection against rabies virus 1 year after vaccination
Vaccine, 2001Co-Authors: Donald L. Lodmell, Larry C. Ewalt, Michael J Parnell, John R Bailey, Cathleen A HanlonAbstract:We have previously shown that Macaca fascicularis (Cynomologus) monkeys receiving a primary and either one or two booster rabies DNA vaccinations are protected against rabies virus. In this study, we determined whether monkeys that had been vaccinated only once via Gene Gun or intramuscularly (i.m.) with different concentrations of DNA would be protected against rabies virus challenge. Neutralizing antibody responses were assayed for 1 year before the monkeys were challenged. Neutralizing antibody was detected at least 50 days earlier in Gene Gun vaccinated as compared to i.m. vaccinated animals. Prior to viral challenge, all (6/6, 100%) Gene Gun vaccinated animals, but only 3/6 (50%) i.m. vaccinated animals seroconverted. In General, antibody titers of the Gene Gun vaccinated animals were higher than the titers of the i.m. vaccinated animals. There was no correlation between the concentration of DNA used for vaccination, the neutralizing antibody responses elicited and protection against viral challenge. Seven days after viral challenge, a rapid and strong anamnestic antibody response was elicited in 100% of the Gene Gun vaccinated monkeys and in four i.m. vaccinated monkeys. Neutralizing antibody remained undetectable in two i.m. vaccinated monkeys. Overall, 60% (3/5) of the Gene Gun vaccinated animals and 87% (5/6) of the i.m. vaccinated monkeys survived viral challenge. This study is the first, to our knowledge, to show long-term protection of non-human primates against a human viral pathogen using a DNA vaccination protocol that did not include a booster immunization.
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Gene Gun particle-mediated vaccination with plasmid DNA confers protective immunity against rabies virus infection
Vaccine, 1998Co-Authors: Donald L. Lodmell, Nancy B. Ray, Larry C. EwaltAbstract:Accell® Gene Gun particle-mediated immunization with DNA encoding the glycoprotein Gene of the challenge virus standard strain of rabies virus was evaluated for its ability to elicit protective levels of serum anti-rabies virus neutralizing antibody. Strong primary and booster neutralizing antibody responses were detected in mice following immunization with 2 μg of DNA coated on 2.6-μm gold beads. Protective levels of antibody persisted for over 300 days. Mice challenged intraplantarly 315 days post-primary immunization (225 days post-booster vaccination) survived lethal rabies virus challenge. Our data demonstrate a potentially significant role for Gene Gun-based delivery of DNA in the field of rabies virus vaccination.
L-c Yang - One of the best experts on this subject based on the ideXlab platform.
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Gene Gun particle encoding preproenkephalin cDNA produces analgesia against capsaicin-induced bladder pain in rats
Urology, 2005Co-Authors: Yaochi Chuang, Pohui Chiang, L-c Yang, Michael B Chancellor, Hong-yo Kang, Wen Lung, Fernando Demiguel, Naoki YoshimuraAbstract:Abstract Objectives To evaluate the efficacy of Gene therapy using a Gene Gun or direct injection for the transfer of human preproenkephalin (PPE) plasmid cDNA using a capsaicin-induced bladder pain model in rats. Opioid peptides play an essential role in the modulation of micturition reflex and control of inflammatory pain. PPE is one such precursor molecule. Methods Human PPE cDNA was cloned into a modified pCMV plasmid and delivered into the bladder wall of adult female rats by direct injection or Gene Gun. At 4 and 7 days after Gene therapy, continuous cystometrograms were performed under urethane anesthesia by filling the bladder (0.08 mL/min) with saline, followed by 15 μM capsaicin. Immunohistochemical staining was used to detect enkephalins in the bladder after PPE cDNA transfer. Results The intercontraction interval was decreased after intravesical instillation of capsaicin (65.0% and 63.1% decrease) in the control group or direct PPE Gene injection group, respectively. However, the Gene Gun-treated group showed a significantly reduced response to capsaicin instillation at day 4 and day 7 (intercontraction interval 16.2% and 42.8% decrease, respectively). This analgesic effect was reversed by intravenous naloxone, an opioid antagonist (5 mg/kg). Increased enkephalin immunoreactivity in the bladder was observed in the Gene Gun-treated group at day 4, which was reduced at day 7. Conclusions The PPE Gene can be effectively transferred and suppress the nociceptive response in the bladder using the Gene Gun method. These results support potential clinical application of PPE Gene Gun delivery system for the treatment of bladder pain and other types of visceral pain.
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Gene therapy for bladder pain with Gene Gun particle encoding pro opiomelanocortin cdna
The Journal of Urology, 2003Co-Authors: Yaochi Chuang, Ankuo Chou, Pohui Chiang, T.j. Yu, P. C. Wu, L-c Yang, Naoki Yoshimura, Michael B ChancellorAbstract:ABSTRACTPurpose: Interstitial cystitis is a bladder hypersensitivity disease associated with bladder pain that has been a major challenge to understand and treat. We hypothesized that targeted and localized expression of endogenous opioid peptide in the bladder could be useful for the treatment of bladder pain. Pro-opiomelanocortin (POMC) is one of such precursor molecules. In this study we developed a Gene Gun method for the transfer of POMC cDNA in vivo and investigated its therapeutic effect on acetic acid induced bladder hyperactivity in rats.Materials and Methods: Human POMC cDNA was cloned into a modified pCMV plasmid and delivered into the bladder wall of adult female rats by direct injection or the Gene Gun. Three days after Gene therapy continuous cystometrograms were performed using urethane anesthesia by filling the bladder (0.08 ml per minute) with saline, followed by 0.3% acetic acid. Bladder immunohistochemical testing was used to detect endorphin after POMC cDNA transfer.Results: The interc...
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Gene-Gun particle with pro-opiomelanocortin cDNA produces analgesia against formalin-induced pain in rats
Gene Therapy, 2002Co-Authors: A-k Chou, C-h Yang, J-t Chen, S-h Lin, R Muhammad, L-c YangAbstract:Endogenous opioid peptides play an essential role in the intrinsic modulation and control of inflammatory pain, and could be therapeutically useful. These opioid peptides are synthesized as parts of larger precursor molecules. One such precursor molecule is pro-opiomelanocortin (POMC). In this study, we developed a Gene-Gun method for the transfer of POMC cDNA in vivo , and investigated its therapeutic effect on inflammatory pain in a rat model of formalin-induced pain. Human POMC cDNA was cloned into a modified pCMV plasmid and delivered to the skin of rats by Gene Gun. Three days after Gene-Gun injection, 1% formalin was injected. Endorphin levels were measured in the serum and skin after the formalin test, and skin histology was used to detect endorphin after green fluorescent protein (GFP; control) or POMC cDNA transfer. There was no significant difference in the results of acute nociceptive tests between the experimental and control groups. There was also no difference in response between the groups to phase 1 of the formalin test. However, rats which received POMC cDNA via Gene-Gun injection showed a significantly reduced response in phase 2 of the formalin test. Endorphin immunoreactivity in the skin increased approximately three- to four-fold in experimental animals compared with GFP-treated controls at day 3 after injection. The phase 2 response in animals treated with formalin and naloxone did not differ significantly from the control, implying that the analgesic effects of POMC cDNA particle injection in phase 2 of the formalin test are reversed by naloxone. There are two major findings from this study. First, in vivo DNA delivery by Gene Gun to the skin is feasible. Second, the production of β-endorphin is insufficient to block phasic pain, but is effective against sensitization of the afferent neurons during phase 2 of the formalin test.
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Gene-Gun particle with pro-opiomelanocortin cDNA produces analgesia against formalin-induced pain in rats
Gene therapy, 2002Co-Authors: Ankuo Chou, C-h Yang, J-t Chen, S-h Lin, R Muhammad, L-c YangAbstract:Endogenous opioid peptides play an essential role in the intrinsic modulation and control of inflammatory pain, and could be therapeutically useful. These opioid peptides are synthesized as parts of larger precursor molecules. One such precursor molecule is pro-opiomelanocortin (POMC). In this study, we developed a Gene-Gun method for the transfer of POMC cDNA in vivo, and investigated its therapeutic effect on inflammatory pain in a rat model of formalin-induced pain. Human POMC cDNA was cloned into a modified pCMV plasmid and delivered to the skin of rats by Gene Gun. Three days after Gene-Gun injection, 1% formalin was injected. Endorphin levels were measured in the serum and skin after the formalin test, and skin histology was used to detect endorphin after green fluorescent protein (GFP; control) or POMC cDNA transfer. There was no significant difference in the results of acute nociceptive tests between the experimental and control groups. There was also no difference in response between the groups to phase 1 of the formalin test. However, rats which received POMC cDNA via Gene-Gun injection showed a significantly reduced response in phase 2 of the formalin test. Endorphin immunoreactivity in the skin increased approximately three- to four-fold in experimental animals compared with GFP-treated controls at day 3 after injection. The phase 2 response in animals treated with formalin and naloxone did not differ significantly from the control, implying that the analgesic effects of POMC cDNA particle injection in phase 2 of the formalin test are reversed by naloxone. There are two major findings from this study. First, in vivo DNA delivery by Gene Gun to the skin is feasible. Second, the production of beta-endorphin is insufficient to block phasic pain, but is effective against sensitization of the afferent neurons during phase 2 of the formalin test.
Donald L. Lodmell - One of the best experts on this subject based on the ideXlab platform.
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one time Gene Gun or intramuscular rabies dna vaccination of non human primates comparison of neutralizing antibody responses and protection against rabies virus 1 year after vaccination
Vaccine, 2001Co-Authors: Donald L. Lodmell, Larry C. Ewalt, Michael J Parnell, John R Bailey, Cathleen A HanlonAbstract:We have previously shown that Macaca fascicularis (Cynomologus) monkeys receiving a primary and either one or two booster rabies DNA vaccinations are protected against rabies virus. In this study, we determined whether monkeys that had been vaccinated only once via Gene Gun or intramuscularly (i.m.) with different concentrations of DNA would be protected against rabies virus challenge. Neutralizing antibody responses were assayed for 1 year before the monkeys were challenged. Neutralizing antibody was detected at least 50 days earlier in Gene Gun vaccinated as compared to i.m. vaccinated animals. Prior to viral challenge, all (6/6, 100%) Gene Gun vaccinated animals, but only 3/6 (50%) i.m. vaccinated animals seroconverted. In General, antibody titers of the Gene Gun vaccinated animals were higher than the titers of the i.m. vaccinated animals. There was no correlation between the concentration of DNA used for vaccination, the neutralizing antibody responses elicited and protection against viral challenge. Seven days after viral challenge, a rapid and strong anamnestic antibody response was elicited in 100% of the Gene Gun vaccinated monkeys and in four i.m. vaccinated monkeys. Neutralizing antibody remained undetectable in two i.m. vaccinated monkeys. Overall, 60% (3/5) of the Gene Gun vaccinated animals and 87% (5/6) of the i.m. vaccinated monkeys survived viral challenge. This study is the first, to our knowledge, to show long-term protection of non-human primates against a human viral pathogen using a DNA vaccination protocol that did not include a booster immunization.
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Gene Gun particle-mediated vaccination with plasmid DNA confers protective immunity against rabies virus infection
Vaccine, 1998Co-Authors: Donald L. Lodmell, Nancy B. Ray, Larry C. EwaltAbstract:Accell® Gene Gun particle-mediated immunization with DNA encoding the glycoprotein Gene of the challenge virus standard strain of rabies virus was evaluated for its ability to elicit protective levels of serum anti-rabies virus neutralizing antibody. Strong primary and booster neutralizing antibody responses were detected in mice following immunization with 2 μg of DNA coated on 2.6-μm gold beads. Protective levels of antibody persisted for over 300 days. Mice challenged intraplantarly 315 days post-primary immunization (225 days post-booster vaccination) survived lethal rabies virus challenge. Our data demonstrate a potentially significant role for Gene Gun-based delivery of DNA in the field of rabies virus vaccination.
Ming Derg Lai - One of the best experts on this subject based on the ideXlab platform.
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Biolistic DNA delivery to mice with the low pressure Gene Gun.
Methods in molecular biology (Clifton N.J.), 2012Co-Authors: Meng-chi Yen, Ming Derg LaiAbstract:Biolistic DNA delivery is an approach to deliver plasmid to culture cells, plants, or animals. Plasmid DNA is usually transferred through bombardment of DNA-coated particles by highly pressurized gas in various kinds of delivery vehicles. The low pressure Gene Gun can deliver plasmid at lower pressure. Here, we describe methods of biolistic DNA delivery to mice using the low pressure Gene Gun.
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752. Delivery of Non-Microparticle Naked DNA Vaccine Using Supersonic Flow by a Low-Pressure Gene Gun
Molecular Therapy, 2006Co-Authors: Chi-chen Lin, Ying-chang Wang, Men-chi Yen, Ming Derg LaiAbstract:DNA vaccines are a new and powerful approach to Generation of immunological responses against infectious disease and cancer. DNA can be delivered either into muscle by simple injection or into epidermal by Gene Gun. Intramuscular injection requires large amount of DNA (100microgram/per mouse) to elicit the immune response; in contrast, microparticlulate bombardment system can induce immune response using very low amount of DNA (1 microgram/per mouse). One disadvantage of Gene Gun bombardment is that non-biodegradable gold or tungsten may skew the immune response or cause adverse side effect when accumulated. In this report, we have demonstrated the direct delivery of naked DNA without any microparticle using a modified Gene Gun. The modified Gene Gun is based on the aerodynamic theory that a supersonic flow is Generated when the pressure difference between the inside and the outside pressure of the nozzle is greater than 1.9atm. This high speed airflow can carry the particle from stationary state to accelerate to an extreme high speed (200m/sec). In this way, the naked DNA may be directly transformed into the mammalian cells. Previous study indicated that this modified Gene Gun achieve similar deliver efficacy as the Gene Gun commercially available using gold-coated DNA. In this report, we showed that the modified Gene Gun can achieve 10|[ndash]|40% delivery efficacy as the gold particle coated with DNA in Balb/C mice, C57BL/6 mice, and C3H mice using luciferase Gene reporter driven by CMV promoter. Then, we examined the immune responses elicited by inoculation of DNA encoding HBV large surface protein expressed by its own promoter. The non-microparticle DNA and gold-coated DNA elicited similar humoral immunity as shown by antibody titer. Similar overall cytokine profile was induced by either type of DNA vaccine. Furthermore, we examined the therapeutic responses with the DNA vaccine against the extracellular domain of neu on the mouse tumor (MBT-2) naturally overexpressing neu in C3H mice. The results indicated that non-microparticle naked neu DNA vaccine can achieve similar anti-tumor effect as the gold-coated neu DNA vaccine in C3H mice at the dose of 1 microgram/per mouse.
Antonio Pezzutto - One of the best experts on this subject based on the ideXlab platform.
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HER2/neu DNA vaccination by intradermal Gene delivery in a mouse tumor model: Gene Gun is superior to jet injector in inducing CTL responses and protective immunity.
Oncoimmunology, 2012Co-Authors: T Nguyen-hoai, Dennis Kobelt, Oliver Hohn, Peter M. Schlag, Bernd Dörken, Steven Norley, Martin Lipp, Wolfgang Walther, Antonio PezzuttoAbstract:DNA vaccines are potential tools for the induction of immune responses against both infectious disease and cancer. The dermal application of DNA vaccines is of particular interest since the epidermal and dermal layers of the skin are characterized by an abundance of antigen-presenting cells (APCs). The aim of our study was to compare tumor protection as obtained by two different methods of intradermal DNA delivery (Gene Gun and jet injector) in a well-established HER2/neu mouse tumor model. BALB/c mice were immunized twice with a HER2/neu-coding plasmid by Gene Gun or jet injector. Mice were then subcutaneously challenged with HER2/neu+ synGeneic D2F2/E2 tumor cells. Protection against subsequent challenges with tumor cells as well as humoral and T-cell immune responses induced by the vaccine were monitored. Gene Gun immunization was far superior to jet injector both in terms of tumor protection and induction of HER2/neu-specific immune responses. After Gene Gun immunization, 60% of the mice remained tumor-free until day 140 as compared with 25% after jet injector immunization. Furthermore, Gene Gun vaccination was able to induce both a strong TH1-polarized T-cell response with detectable cytotoxic T-lymphocyte (CTL) activity and a humoral immune response against HER2/neu, whereas the jet injector was not. Although the disadvantages that were associated with the use of the jet injector in our model may be overcome with methodological modifications and/or in larger animals, which exhibit a thicker skin and/or subcutaneous muscle tissue, we conclude that Gene Gun delivery constitutes the method of choice for intradermal DNA delivery in preclinical mouse models and possibly also for the clinical development of DNA-based vaccines.
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her2 neu dna vaccination by intradermal Gene delivery in a mouse tumor model Gene Gun is superior to jet injector in inducing ctl responses and protective immunity
OncoImmunology, 2012Co-Authors: T Nguyenhoai, Dennis Kobelt, Oliver Hohn, Peter M. Schlag, Bernd Dörken, Steven Norley, Martin Lipp, Wolfgang Walther, Antonio Pezzutto, Jorg WestermannAbstract:DNA vaccines are potential tools for the induction of immune responses against both infectious disease and cancer. The dermal application of DNA vaccines is of particular interest since the epidermal and dermal layers of the skin are characterized by an abundance of antigen-presenting cells (APCs). The aim of our study was to compare tumor protection as obtained by two different methods of intradermal DNA delivery (Gene Gun and jet injector) in a well-established HER2/neu mouse tumor model. BALB/c mice were immunized twice with a HER2/neu-coding plasmid by Gene Gun or jet injector. Mice were then subcutaneously challenged with HER2/neu+ synGeneic D2F2/E2 tumor cells. Protection against subsequent challenges with tumor cells as well as humoral and T-cell immune responses induced by the vaccine were monitored. Gene Gun immunization was far superior to jet injector both in terms of tumor protection and induction of HER2/neu-specific immune responses. After Gene Gun immunization, 60% of the mice remained tumor-free until day 140 as compared with 25% after jet injector immunization. Furthermore, Gene Gun vaccination was able to induce both a strong TH1-polarized T-cell response with detectable cytotoxic T-lymphocyte (CTL) activity and a humoral immune response against HER2/neu, whereas the jet injector was not. Although the disadvantages that were associated with the use of the jet injector in our model may be overcome with methodological modifications and/or in larger animals, which exhibit a thicker skin and/or subcutaneous muscle tissue, we conclude that Gene Gun delivery constitutes the method of choice for intradermal DNA delivery in preclinical mouse models and possibly also for the clinical development of DNA-based vaccines.