A Multi-Component Prime-Boost Vaccination Regimen with a Consensus MOMP Antigen Enhances Chlamydia trachomatis Clearance

The Supplementary Material for this article can be found online at http://journal.frontiersin.org/article/10.3389/fimmu.2016.00162BACKGROUND: A vaccine for Chlamydia trachomatis is of urgent medical need. We explored bioinformatic approaches to generate an immunogen against C. trachomatis that would induce cross-serovar T-cell responses as (i) CD4(+) T cells have been shown in animal models and human studies to be important in chlamydial protection and (ii) antibody responses may be restrictive and serovar specific. METHODS: A consensus antigen based on over 1,500 major outer membrane protein (MOMP) sequences provided high epitope coverage against the most prevalent C. trachomatis strains in silico. Having designed the T-cell immunogen, we assessed it for immunogenicity in prime-boost regimens. This consensus MOMP transgene was delivered using plasmid DNA, Human Adenovirus 5 (HuAd5) or modified vaccinia Ankara (MVA) vectors with or without MF59(®) adjuvanted recombinant MOMP protein. RESULTS: Different regimens induced distinct immune profiles. The DNA-HuAd5-MVA-Protein vaccine regimen induced a cellular response with a Th1-biased serum antibody response, alongside high serum and vaginal MOMP-specific antibodies. This regimen significantly enhanced clearance against intravaginal C. trachomatis serovar D infection in both BALB/c and B6C3F1 mouse strains. This enhanced clearance was shown to be CD4(+) T-cell dependent. Future studies will need to confirm the specificity and precise mechanisms of protection. CONCLUSION: A C. trachomatis vaccine needs to induce a robust cellular response with broad cross-serovar coverage and a heterologous prime-boost regimen may be an approach to achieve this.AB was funded by the Wellcome Trust. RS was supported by the European Community’s European 7th Framework Program ADITEC (HEALTH-F4-2011-18 280873).info:eu-repo/semantics/publishedVersio

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..

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

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

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

Integrative Biology Approach Identifies Cytokine Targeting Strategies for Psoriasis

Cytokines are critical checkpoints of inflammation. The treatment of human autoimmune disease has been revolutionized by targeting inflammatory cytokines as key drivers of disease pathogenesis. Despite this, there exist numerous pitfalls when translating preclinical data into the clinic. We developed an integrative biology approach combining human disease transcriptome data sets with clinically relevant in vivo models in an attempt to bridge this translational gap. We chose interleukin-22 (IL-22) as a model cytokine because of its potentially important proinflammatory role in epithelial tissues. Injection of IL-22 into normal human skin grafts produced marked inflammatory skin changes resembling human psoriasis. Injection of anti-IL-22 monoclonal antibody in a human xenotransplant model of psoriasis, developed specifically to test potential therapeutic candidates, efficiently blocked skin inflammation. Bioinformatic analysis integrating both the IL-22 and anti-IL-22 cytokine transcriptomes and mapping them onto a psoriasis disease gene coexpression network identified key cytokine-dependent hub genes. Using knockout mice and small-molecule blockade, we show that one of these hub genes, the so far unexplored serine/threonine kinase PIM1, is a critical checkpoint for human skin inflammation and potential future therapeutic target in psoriasis. Using in silico integration of human data sets and biological models, we were able to identify a new target in the treatment of psoriasis

Resident CD141 (BDCA3)(+) dendritic cells in human skin produce IL-10 and induce regulatory T cells that suppress skin inflammation

Human skin immune homeostasis, and its regulation by specialized subsets of tissue-residing immune sentinels, is poorly understood. In this study, we identify an immunoregulatory tissue-resident dendritic cell (DC) in the dermis of human skin that is characterized by surface expression of CD141, CD14, and constitutive IL-10 secretion (CD141(+) DDCs). CD141(+) DDCs possess lymph node migratory capacity, induce T cell hyporesponsiveness, cross-present self-antigens to autoreactive T cells, and induce potent regulatory T cells that inhibit skin inflammation. Vitamin D(3) (VitD3) promotes certain phenotypic and functional properties of tissue-resident CD141(+) DDCs from human blood DCs. These CD141(+) DDC-like cells can be generated in vitro and, once transferred in vivo, have the capacity to inhibit xeno-graft versus host disease and tumor alloimmunity. These findings suggest that CD141(+) DDCs play an essential role in the maintenance of skin homeostasis and in the regulation of both systemic and tumor alloimmunity. Finally, VitD3-induced CD141(+) DDC-like cells have potential clinical use for their capacity to induce immune tolerance