DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants

Tattooing is one of a number of DNA delivery methods which results in an efficient expression of an introduced gene in the epidermal and dermal layers of the skin. The tattoo procedure causes many minor mechanical injuries followed by hemorrhage, necrosis, inflammation and regeneration of the skin and thus non-specifically stimulates the immune system. DNA vaccines delivered by tattooing have been shown to induce higher specific humoral and cellular immune responses than intramuscularly injected DNA. In this study, we focused on the comparison of DNA immunization protocols using different routes of administrations of DNA (intradermal tattoo versus intramuscular injection) and molecular adjuvants (cardiotoxin pre-treatment or GM-CSF DNA co-delivery). For this comparison we used the major capsid protein L1 of human papillomavirus type 16 as a model antigen. L1-specific immune responses were detected after three and four immunizations with 50 μg plasmid DNA. Cardiotoxin pretreatment or GM-CSF DNA co-delivery substantially enhanced the efficacy of DNA vaccine delivered intramuscularly by needle injection but had virtually no effect on the intradermal tattoo vaccination. The promoting effect of both adjuvants was more pronounced after three rather than four immunizations. However, three DNA tattoo immunizations without any adjuvant induced significantly higher L1-specific humoral immune responses than three or even four intramuscular DNA injections supported by molecular adjuvants. Tattooing also elicited significantly higher L1-specific cellular immune responses than intramuscularly delivered DNA in combination with adjuvants. In addition, the lymphocytes of mice treated with the tattoo device proliferated more strongly after mitogen stimulation suggesting the presence of inflammatory responses after tattooing. The tattoo delivery of DNA is a cost-effective method that may be used in laboratory conditions when more rapid and more robust immune responses are required

The use of a silicone-based biomembrane for microaerobic H2S removal from biogas

A lab-scale bio-membrane unit was developed to improve H2S removal from biogas through microaeration. Biomembrane separated biogas from air and consisted of a silicone tube covered by microaerobic biofilm. This setup allowed efficient H2S removal while minimizing biogas contamination with oxygen and nitrogen. The transport and removal of H2S, N-2, O-2, CH4 and CO2 through bare membrane, wet membrane and biomembrane was investigated. Membrane allowed the transfer of gases through it as long as there was enough driving force to induce it. H2S concentration in biogas decreased much faster with the biomembrane. The permeation of gases through the membranes decreased in order: H2S > CO2 > CH4 > O-2 > N-2. H2S removal efficiency of more than 99% was observed during the continuous experiment. Light yellow deposits on the membrane indicated the possible elemental sulfur formation due to biological oxidation of H2S. Thiobacillus thioparus was detected by FISH and PCR-DGGE

Deep Sclerectomy with Nonabsorbable Implant (T-Flux) in Patients with Pseudoexfoliation Glaucoma

Purpose. To evaluate the effectiveness of deep sclerectomy with T-flux implant (DS T-flux) in patients with pseudoexfoliation glaucoma (PExG). Methods. 20 eyes of 18 patients with medically uncontrolled PExG have undergone DS T-flux implantation. Postoperatively we evaluated the IOP values and the frequency of complications. The minimum follow-up time was 12 months (20 eyes) and the maximum 24 months (10 eyes). Results. The mean preoperative IOP was 36.8±8.7 mmHg. The IOP significantly decreased throughout all postoperative periods (P<0.05) and reached 1 day after surgery 11.45±6.6 mmHg; 3 months 13.45±3.6 mmHg; 12 months 14±2.8 mmHg; and 24 months 14.80±2.4 mmHg. Complete success rate, defined as IOP ≤ 18 mmHg without medication, was 85% (17/20 eyes) at 12 months. Qualified success rate, defined as IOP ≤ 18 mmHg with or without medication, was 100% (20/20 eyes). The most frequent postoperative complications were mild hyphaema (9 patients, 45%), choroidal detachment (3 patients, 15%), and hypotony—IOP < 5 mmHg (2 patients, 10%). Conclusions. DS with T-flux implant is a safe and effective surgical treatment method for medically uncontrolled PExG. The number of complications is low

DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants-2

28 and 98 either by tattoo or intramuscular delivery without any adjuvant, in combination with prior application of cardiotoxin 5 days before the first immunization or in mixture with mouse GM-CSF DNA (1:1). A control group of three mice was tattooed with mouse GM-CSF DNA. Splenocytes were isolated 9 days after the last DNA immunization and examined in 4 serial dilutions in the IFN-γ-ELISPOT assay. The representative numbers of spots reflecting IFN-γ-producing cells per 250,000 splenocytes are shown. Splenocytes were stimulated non-specifically with mitogen or specifically with the L1 peptide (aa 165–173). Non-stimulated splenocytes were used as negative controls.<p><b>Copyright information:</b></p><p>Taken from "DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants"</p><p>http://www.gvt-journal.com/content/6/1/4</p><p>Genetic Vaccines and Therapy 2008;6():4-4.</p><p>Published online 7 Feb 2008</p><p>PMCID:PMC2267179.</p><p></p

DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants-0

T, in combination with prior application of cardiotoxin or in mixture with mouse GM-CSF DNA (ratio 1:1). For control, a group of mice was tattooed with mouse GM-CSF DNA. The blood was collected after 3 and 4 immunizations for the estimation of L1-specific antibodies. The end-point titration of sera was performed. The titers of L1-antibodies were determined using an absorption value of 0.4 as cut-off for ELISA.<p><b>Copyright information:</b></p><p>Taken from "DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants"</p><p>http://www.gvt-journal.com/content/6/1/4</p><p>Genetic Vaccines and Therapy 2008;6():4-4.</p><p>Published online 7 Feb 2008</p><p>PMCID:PMC2267179.</p><p></p

DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants-1

intramuscularly after cardiotoxin pre-treatment or by tattoo. Mice immunized intramuscularly with HPV 16 L1 DNA after cardiotoxin pretreatment developed lower levels of L1-specific antibodies than L1-tattooed mice. Serum values below the ELISA cut-off value of 0.4 optical density (O.D.) at 405 nm were considered to be negative.<p><b>Copyright information:</b></p><p>Taken from "DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants"</p><p>http://www.gvt-journal.com/content/6/1/4</p><p>Genetic Vaccines and Therapy 2008;6():4-4.</p><p>Published online 7 Feb 2008</p><p>PMCID:PMC2267179.</p><p></p

DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants-3

T, in combination with prior application of cardiotoxin or in mixture with mouse GM-CSF DNA (ratio 1:1). For control, a group of mice was tattooed with mouse GM-CSF DNA. The blood was collected after 3 and 4 immunizations for the estimation of L1-specific antibodies. The end-point titration of sera was performed. The titers of L1-antibodies were determined using an absorption value of 0.4 as cut-off for ELISA.<p><b>Copyright information:</b></p><p>Taken from "DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants"</p><p>http://www.gvt-journal.com/content/6/1/4</p><p>Genetic Vaccines and Therapy 2008;6():4-4.</p><p>Published online 7 Feb 2008</p><p>PMCID:PMC2267179.</p><p></p

DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants-4

intramuscularly after cardiotoxin pre-treatment or by tattoo. Mice immunized intramuscularly with HPV 16 L1 DNA after cardiotoxin pretreatment developed lower levels of L1-specific antibodies than L1-tattooed mice. Serum values below the ELISA cut-off value of 0.4 optical density (O.D.) at 405 nm were considered to be negative.<p><b>Copyright information:</b></p><p>Taken from "DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants"</p><p>http://www.gvt-journal.com/content/6/1/4</p><p>Genetic Vaccines and Therapy 2008;6():4-4.</p><p>Published online 7 Feb 2008</p><p>PMCID:PMC2267179.</p><p></p