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

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

Effect of PSV-serum premix incubation time on neutralization titer.

<p>A serum from a Gardasil® vaccinated individual was pre-incubated for different times with HPV 16 or HPV 18 PSVs before the neutralization assay was initiated by the addition of reporter cells. ED₅₀ values with 95% confidence intervals are shown.</p

Titration of vaccine sera in the HT-PBNA using PSV of HPV types 16, 18, 31, 45, 52, 58 and BPV-1. A

<p>) Standard serum from Gardasil® vaccinated person. <b>B</b>) Cervarix® vaccine serum. <b>C</b>) Serum from a mouse immunized with L2 epitopes (amino acids 17–36) from HPV16 inserted in the capsid of adeno-associated virus 2 particles.</p

Analytical sensitivity and type-specificity of the HPV 16 and HPV 18 HT-PBNA.

<p>Titration of the WHO International Standards for antibodies to HPV 16 (left) and HPV 18 (right) in HT-PBNA using PSV of HPV types 16, 18, 31, 45, 52, 58 and BPV-1.</p

Comparison of HPV 16 HT-PBNA (Gaussia) with manPBNA (SEAP and Gaussia) and GST-HPV 16 L1 antibody binding assay for natural and vaccine-induced HPV16-specific responses.

<p>HPV 16 HT-PBNA (A–D), manPBNA using SEAP reporter (A and B; titers, ED₅₀) and a bead-based GST-HPV 16 L1 antibody binding assay (C and D; median fluorescent intensity (MFI) at 1∶100 or 1∶2700 serum dilution) were used to determine reactivity of pre- (n = 35; A and C) and post-vaccination (n = 72; B and D) sera. Serum samples analyzed were from women with HPV 16 positive, high-grade cervical intraepithelial neoplasia (base-line sera of the chimeric HPV 16 L1-E7 vaccination trial). Cut off values used for positive/negative classification (broken lines in C) were an ED₅₀ of 80 for the HT-PBNA and 109 MFI at 1∶100 for the GST-L1 antibody binding assay.</p

Detection of neutralizing antibodies as result of natural infection by HT-PBNA.

<p>Thirty-five pre-vaccination sera from a study involving patients with CIN2+ lesions were tested for neutralizing antibodies against PSVs of HPV types 16, 18 and 31. The geometrical mean titer for each HPV type is indicated as a horizontal line and the cut off value (ED₅₀ = 80) as a dashed line.</p