Sera from each group of animals were tested in a functional HIV neutralising TZM-bl assay.

<p>The serum dilution that was able to achieve half maximal inhibition of HIV infectivity was higher in Group B (3 × ID + 2 × IN) for both a closely sequence matched Clade C virus CN54 (A) and a more sequence divergent Clade C HIV (B). All animals from this group achieved neutralisation compared to only 4 of the 7 animals in Group A (3 × IN + 2 × ID) (C).</p

Determination of paternity

The primary theme of the thesis is "paternity determination", an interesting part of private law. The goal of the thesis was to find juridical legislation that could be applied by legal institutions in the Czech Republic, and have been neither legally controlled nor properly spoken through. Also the thesis deals with surrogate motherhood institutes, same-sex parenthood, baby hatches and paternity determination of children, made by assisted reproduction. The thesis is divided into five chapters, which are divided further into sub-chapters. Following the introduction, the primary terms of parental difficulties, paternity determination and the term family are explored. The second chapter discusses motherhood institutes. It speaks primarily of surrogate motherhood's challenges, including other countries' attitude to this topic. The following chapter is about fatherhood determination, containing the juridical legislative analysis of the problem in the Czech law. The fourth chapter expands on the issues of same sex couple parenthood, and the forms of family coexistence. These issues are described in great detail from both the view of Czech law, and also as a global issue. The last chapter discusses the issues of baby hatches, and legislation regarding anonymity for those who utilize them in the Czech..

Spearman Rank correlations comparing serum, nasal and vaginal mucosal IgG.

<p>A strong positive correlation was found at week 9 and 12 between the amounts of antigen-specific antibody present at each site but this relationship was lost by the end of the vaccination regimen as the nasal compartment exhibited increased levels of specific IgG while vaccine-specific antibody decreased in the serum and vaginal vault. By week 15, 3 weeks after the final vaccinations, the IgG present in the serum and vaginal samples still correlate strongly while that in the nasal compartment exhibits little or no significant relationship.</p

The antigen-specific polyclonal sera from the different regimens exhibited differing avidity indices throughout the course of the vaccinations.

<p>Low avidity = <25%, medium avidity = 25–50% and high avidity = >50%. After two priming vaccinations (week 35) the sera from animals inoculated via the IN route had low avidity binding to the vaccine antigen while animals that received ID injections had medium–high avidity gp140-antigen reactive sera. Optimally adjuvanted ID injections were able to continue to enhance avidity while the IN inoculation failed to enhance or even maintain the previous high level avidity elicited by prior ID injections.</p

Comparison of combined R848 and titrated GLA-AF adjuvants.

<p>A constant quantity of the R848 adjuvant was administered via either the IN or ID routes together with increasing quantities of the GLA-AF adjuvant in final administration volumes of 100 μl.</p

Pre-clinical development of a vaccine for human lymphatic filariasis

This study was conducted to optimize a fusion protein vaccine for translational development as a vaccine against the human tropical parasitic infection, lymphatic filariasis (LF). The vaccine candidate, His-tagged rBmHAXT was developed previously in our laboratory and was tested in various animal models including mouse, gerbils and Rhesus macaque where it exhibited significant levels of vaccine-induced protection. However, for commercial manufacturing and for regulatory approval for human use, there was a need to modify the vaccine antigen and its production and analytical release methods. Therefore, the major focus of this study was to develop a process for manufacturing an affinity tag-free rBmHAXT and evaluate its immunogenicity, potency and protective efficacy in both inbred and outbred mouse models, as well as in outbred gerbil models. Our results demonstrate that the tag-free rBmHAXT vaccine produced with a process suitable for cGMP production had protective properties equivalent to the original His-tagged rBmHAXT

Transcriptomic comparison for five different Sm-p80 vaccine formulations.

<p><b>(A)</b> Distribution of differentially expressed mouse splenocyte genes (y axis) according to ontology classification for mice immunized with different Sm-p80 vaccine formulations and challenged with <i>S</i>. <i>mansoni</i> compared to control challenge in naïve mice (x axis). <b>(B)</b> Comparative levels of differentially expressed genes between formulations. <b>(C)</b> Heat map for relative gene expression values of 59 common genes (rows) comparing <i>S</i>. <i>mansoni</i> challenged and Sm-p80 formulations (columns). Identified genes were statistically significant (<i>P</i> < 0.01, Student’s <i>t</i>-test and greater than 2-fold change with fold change in Log₂). <b>(D)</b>. Heat map showing expression differences of 24 identified genes (rows) unique for Sm-p80 formulations (columns). <b>(E)</b> Circular visualization (CIRCOS) plot showing shared canonical pathways across vaccine formulations and control challenged mice. Significant canonical pathways (<i>P</i>-value <0.05, right-tailed Fisher Exact Test) were identified and compared across each condition.</p

Classical B cell response gene network.

<p>Classical B cell response related gene network with overlaid expression values and z-score of activation.</p

Workflow schematic to evaluate Sm-p80 vaccine efficacy in mouse and baboon models.

<p><b>(A)</b> Experimental design to evaluate five Sm-p80 vaccine formulations for protection against <i>Schistosoma mansoni</i> challenge using transcriptomics. Thirty mice for each vaccination strategy (n = 15 for control and experimental) were immunized with DNA based vaccine (VR1020-Sm-p80); prime-boost approach (primed with DNA vaccine pcDNA3-Sm-p80 and boosted with recombinant Sm-p80 in ODN adjuvant); or with recombinant protein based vaccine (rSm-p80) in different adjuvants (alum, ODN, or GLA). Vaccinated control animals received empty DNA vector or adjuvant alone. All vaccinated animals received two booster immunizations prior to challenge infection with <i>S</i>. <i>mansoni</i>. For control challenge mice, naïve animals were infected with <i>S</i>. <i>mansoni</i> (not shown in figure). Eight weeks post-infection, animals were euthanized and necropsied to assess worm burden protection and tissue collection. Splenocytes from individual mice were pooled for RNA extraction and construction of cDNA libraries for high-throughput sequencing. Bioinformatics and transcriptomics allowed Sm-p80 vaccine formulations comparisons to identify signature molecules and network pathways. <b>(B)</b> Experimental design to identify early-gene signatures during the immune response to rSm-p80 + ODN vaccine. Experimental mice (n = 20) were immunized with rSm-80 + ODN while control mice were injected with ODN alone (n = 20). Five animals per group were euthanized at 24 hours, 48 hours, 7 days, and 21 days post-immunization. Splenocyte RNA was isolated and individual mouse cDNA libraries constructed for high-throughput sequencing. Bioinformatics analysis identified gene-networks during the immune response over time to vaccination with rSm-p80 + ODN. <b>(C)</b> Experimental design to assess rSm-p80 + ODN vaccine efficacy in a non-human primate model using transcriptomics. Baboons immunized with rSm-p80 + ODN (n = 8) or adjuvant alone (n = 8) received two booster vaccinations at four week intervals each. Prior to <i>S</i>. <i>mansoni</i> challenge infection (week 12), peripheral blood mononuclear cells were collected to establish a baseline immune signature (distinguish between vaccinated and control). Eight weeks post-infection (week 20) animals were euthanized and necropsied to assess worm burden. Individual tissue samples (PBMCs, spleens, and mesenteric lymph nodes) from each baboon were processed for RNA extraction and library preparation for sequencing. Data analysis identified Sm-p80 induced baboon tissue specific gene networks.</p

Cellular functions and inflammatory response network.

<p>IPA identified cellular functions and inflammatory response network at 24 hours post-vaccination. Expression values for 21 days, rSm-p80 + ODN post-challenge, and <i>S</i>. <i>mansoni</i> control challenge datasets were overlaid with predicted activation scores.</p