Blocking Junctional Adhesion Molecule C Enhances Dendritic Cell Migration and Boosts the Immune Responses against Leishmania major.

The recruitment of dendritic cells to sites of infections and their migration to lymph nodes is fundamental for antigen processing and presentation to T cells. In the present study, we showed that antibody blockade of junctional adhesion molecule C (JAM-C) on endothelial cells removed JAM-C away from junctions and increased vascular permeability after L. major infection. This has multiple consequences on the output of the immune response. In resistant C57BL/6 and susceptible BALB/c mice, we found higher numbers of innate immune cells migrating from blood to the site of infection. The subsequent migration of dendritic cells (DCs) from the skin to the draining lymph node was also improved, thereby boosting the induction of the adaptive immune response. In C57BL/6 mice, JAM-C blockade after L. major injection led to an enhanced IFN-γ dominated T helper 1 (Th1) response with reduced skin lesions and parasite burden. Conversely, anti JAM-C treatment increased the IL-4-driven T helper 2 (Th2) response in BALB/c mice with disease exacerbation. Overall, our results show that JAM-C blockade can finely-tune the innate cell migration and accelerate the consequent immune response to L. major without changing the type of the T helper cell response.

The contributions of sensory dominance and attentional bias to cross-modal enhancement of visual cortex excitability.

Approaching or looming sounds (L-sounds) have been shown to selectively increase visual cortex excitability [Romei, V., Murray, M. M., Cappe, C., & Thut, G. Preperceptual and stimulus-selective enhancement of low-level human visual cortex excitability by sounds. Current Biology, 19, 1799-1805, 2009]. These cross-modal effects start at an early, preperceptual stage of sound processing and persist with increasing sound duration. Here, we identified individual factors contributing to cross-modal effects on visual cortex excitability and studied the persistence of effects after sound offset. To this end, we probed the impact of different L-sound velocities on phosphene perception postsound as a function of individual auditory versus visual preference/dominance using single-pulse TMS over the occipital pole. We found that the boosting of phosphene perception by L-sounds continued for several tens of milliseconds after the end of the L-sound and was temporally sensitive to different L-sound profiles (velocities). In addition, we found that this depended on an individual's preferred sensory modality (auditory vs. visual) as determined through a divided attention task (attentional preference), but not on their simple threshold detection level per sensory modality. Whereas individuals with "visual preference" showed enhanced phosphene perception irrespective of L-sound velocity, those with "auditory preference" showed differential peaks in phosphene perception whose delays after sound-offset followed the different L-sound velocity profiles. These novel findings suggest that looming signals modulate visual cortex excitability beyond sound duration possibly to support prompt identification and reaction to potentially dangerous approaching objects. The observed interindividual differences favor the idea that unlike early effects this late L-sound impact on visual cortex excitability is influenced by cross-modal attentional mechanisms rather than low-level sensory processes.

Influenza vaccination in immunocompromised patients: efficacy and safety.

Yearly administration of the influenza vaccine is the main strategy to prevent influenza in immunocompromised patients. Here, we reviewed the recent literature regarding the clinical significance of the influenza virus infection, as well as the immunogenicity and safety of the influenza vaccine in HIV‑infected individuals, solid-organ and stem-cell transplant recipients and patients receiving biological agents. Epidemiological data produced during the 2009 influenza pandemic have confirmed that immunocompromised patients remain at high risk of influenza-associated complications, namely viral and bacterial pneumonia, hospitalization and even death. The immunogenicity of the influenza vaccine is overall reduced in immunocompromised patients, although a significant clinical protection from influenza is expected to be obtained with vaccination. Influenza vaccination is safe in immunocompromised patients. The efficacy of novel strategies to improve the immunogenicity to the vaccine, such as the use of adjuvanted vaccines, boosting doses and intradermal vaccination, needs to be validated in appropriately powered clinical trials.

Ensemble methods for environmental data modelling with support vector regression

This paper investigates the use of ensemble of predictors in order to improve the performance of spatial prediction methods. Support vector regression (SVR), a popular method from the field of statistical machine learning, is used. Several instances of SVR are combined using different data sampling schemes (bagging and boosting). Bagging shows good performance, and proves to be more computationally efficient than training a single SVR model while reducing error. Boosting, however, does not improve results on this specific problem.

Low TCR avidity and lack of tumor cell recognition in CD8(+) T cells primed with the CEA-analogue CAP1-6D peptide.

The use of "altered peptide ligands" (APL), epitopes designed for exerting increased immunogenicity as compared with native determinants, represents nowadays one of the most utilized strategies for overcoming immune tolerance to self-antigens and boosting anti-tumor T cell-mediated immune responses. However, the actual ability of APL-primed T cells to cross-recognize natural epitopes expressed by tumor cells remains a crucial concern. In the present study, we show that CAP1-6D, a superagonist analogue of a carcinoembriyonic antigen (CEA)-derived HLA-A*0201-restricted epitope widely used in clinical setting, reproducibly promotes the generation of low-affinity CD8(+) T cells lacking the ability to recognized CEA-expressing colorectal carcinoma (CRC) cells. Short-term T cell cultures, obtained by priming peripheral blood mononuclear cells from HLA-A*0201(+) healthy donors or CRC patients with CAP1-6D, were indeed found to heterogeneously cross-react with saturating concentrations of the native peptide CAP1, but to fail constantly lysing or recognizing through IFN- gamma release CEA(+)CRC cells. Characterization of anti-CAP1-6D T cell avidity, gained through peptide titration, CD8-dependency assay, and staining with mutated tetramers (D227K/T228A), revealed that anti-CAP1-6D T cells exerted a differential interaction with the two CEA epitopes, i.e., displaying high affinity/CD8-independency toward the APL and low affinity/CD8-dependency toward the native CAP1 peptide. Our data demonstrate that the efficient detection of self-antigen expressed by tumors could be a feature of high avidity CD8-independent T cells, and underline the need for extensive analysis of tumor cross-recognition prior to any clinical usage of APL as anti-cancer vaccines.

IL-12-mediated STAT4 signaling and TCR signal strength cooperate in the induction of CD40L in human and mouse CD8(+) T cells.

CD40L is one of the key molecules bridging the activation of specific T cells and the maturation of professional and nonprofessional antigen-presenting cells including B cells. CD4(+) T cells have been regarded as the major T-cell subset that expresses CD40L upon cognate activation; however, we demonstrate here that a putative CD8(+) helper T-cell subset expressing CD40L is induced in human and murine CD8(+) T cells in vitro and in mice immunized with antigen-pulsed dendritic cells. IL-12 and STAT4-mediated signaling was the major instructive cytokine signal boosting the ability of CD8(+) T cells to express CD40L both in vitro and in vivo. Additionally, TCR signaling strength modulated CD40L expression in CD8(+) T cells after primary differentiation in vitro as well as in vivo. The induction of CD40L in CD8(+) T cells regulated by IL-12 and TCR signaling may enable CD8(+) T cells to respond autonomously of CD4(+) T cells. Thus, we propose that under proinflammatory conditions, a self-sustaining positive feedback loop could facilitate the efficient priming of T cells stimulated by high affinity peptide displaying APCs.

A novel HIV vaccine adjuvanted by IC31 induces robust and persistent humoral and cellular immunity.

The HIV vaccine strategy that, to date, generated immune protection consisted of a prime-boost regimen using a canarypox vector and an HIV envelope protein with alum, as shown in the RV144 trial. Since the efficacy was weak, and previous HIV vaccine trials designed to generate antibody responses failed, we hypothesized that generation of T cell responses would result in improved protection. Thus, we tested the immunogenicity of a similar envelope-based vaccine using a mouse model, with two modifications: a clade C CN54gp140 HIV envelope protein was adjuvanted by the TLR9 agonist IC31®, and the viral vector was the vaccinia strain NYVAC-CN54 expressing HIV envelope gp120. The use of IC31® facilitated immunoglobulin isotype switching, leading to the production of Env-specific IgG2a, as compared to protein with alum alone. Boosting with NYVAC-CN54 resulted in the generation of more robust Th1 T cell responses. Moreover, gp140 prime with IC31® and alum followed by NYVAC-CN54 boost resulted in the formation and persistence of central and effector memory populations in the spleen and an effector memory population in the gut. Our data suggest that this regimen is promising and could improve the protection rate by eliciting strong and long-lasting humoral and cellular immune responses.

Preclinical evaluation of the immunogenicity of C-type HIV-1-based DNA and NYVAC vaccines in the Balb/C mouse model.

As part of a European initiative (EuroVacc), we report the design, construction, and immunogenicity of two HIV-1 vaccine candidates based on a clade C virus strain (CN54) representing the current major epidemic in Asia and parts of Africa. Open reading frames encoding an artificial 160-kDa GagPolNef (GPN) polyprotein and the external glycoprotein gp120 were fully RNA and codon optimized. A DNA vaccine (DNA-GPN and DNA-gp120, referred to as DNA-C), and a replication-deficient vaccinia virus encoding both reading frames (NYVAC-C), were assessed regarding immunogenicity in Balb/C mice. The intramuscular administration of both plasmid DNA constructs, followed by two booster DNA immunizations, induced substantial T-cell responses against both antigens as well as Env-specific antibodies. Whereas low doses of NYVAC-C failed to induce specific CTL or antibodies, high doses generated cellular as well as humoral immune responses, but these did not reach the levels seen following DNA vaccination. The most potent immune responses were detectable using prime:boost protocols, regardless of whether DNA-C or NYVAC-C was used as the priming or boosting agent. These preclinical findings revealed the immunogenic response triggered by DNA-C and its enhancement by combining it with NYVAC-C, thus complementing the macaque preclinical and human phase I clinical studies of EuroVacc.

The three main stumbling blocks for anticancer T cells.

Memory and effector T cells have the potential to counteract cancer progression, but often fail to control the disease, essentially because of three main stumbling blocks. First, clonal deletion leads to relatively low numbers or low-to-intermediate T cell receptor (TCR) affinity of self/tumor-specific T cells. Second, the poor innate immune stimulation by solid tumors is responsible for inefficient priming and boosting. Third, T cells are suppressed in the tumor microenvironment by inhibitory signals from other immune cells, stroma and tumor cells, which induces T cell exhaustion, as demonstrated in metastases of melanoma patients. State-of-the-art adoptive cell transfer and active immunotherapy can partially overcome the three stumbling blocks. The reversibility of T cell exhaustion and novel molecular insights provide the basis for further improvements of clinical immunotherapy.

Suunnittelun tunnusluvut

Diplomityön tavoitteena oli kehittää Andritzin Kuitulinja-divisioonalle tunnuslukuja, työkaluja sekä menetelmiä projektin johdon jasuunnittelun eri osa-alueiden arviointiin, hinnoitteluun ja seurantaan Työn teoriaosuudessa käsiteltiin projektiliiketoimintaa, suunnitteluaja sen hankintaa, tunnuslukuja sekä mittaamista projektiympäristössä. Työn pääpaino oli kuitenkin empiriaosuudella, jossa aluksi tutustuttiin yksityiskohtaisesti työssä tutkittaviin osa-alueisiin. Tiedonkeruuvaiheessa kerättiin suuri määrätunnuslukujen laskemiseksi tarvittavaa tietoa toteutetuista tai vielä hieman keskeneräisistä projekteista. Työn kirjallinen osuus ja kerätty numeerinen tieto yhdistettiin tunnuslukujen analysointivaiheessa, jossa tutkittiin eri tunnuslukujen soveltuvuutta mittaamiseen. Tunnusluvut soveltuivat hyvin joidenkin osa-alueiden kuten prosessi- ja putkistosuunnittelun mittaamiseen. Joidenkin osa-alueiden, kuten rakennussuunnittelun mittaaminen tunnuslukujen avulla oli vaikeampaa tunnuslukujen suuresta hajonnasta johtuen. Eri osa-alueille saatiin kuitenkin määritettyä parhaat tunnusluvut, projektien luokitteluperusteet ja standardiarvot. Työn lopputuloksina saatiin päätavoitteena ollut tunnuslukumittaristo sekä mittaamisen yhtenäistämiseksi luotu tiedonkeruulomake. Mittaamisen tehostamiseksi ja kehittämiseksi tehtiin useita jatkotutkimus- ja kehitysehdotuksia. Lisäksi työssä esitettiin uusi menetelmä projektinohjauksen ja resurssisuunnittelun tueksi.

Viranomaisvaatimusten mukaisten putkistojen hankinta, asennus ja korjaus

Tämän työn tarkoituksena oli kirjallisuustutkimuksen ja aikaisemmin valmistuneiden putkitöiden avulla määritellä ohjeistus viranomaisvaatimusten mukaisten putkistojen valmistamista varten. Tarkasteluesimerkkeinä oli kolme erilaista putkityötä. Näiden seikkojen kautta vahvistui käsitys ohjeistuksen tarpeellisuudesta, mikä oli jo työn tavoitteissa yhtenä päämääränä. Aluksi selvitetään lainsäädäntöä ja sitä, mitä laki vaati putkistoilta. Seuraavaksi tarkastellaan standardeja valmistuksen näkökannalta ja siitä seurauksena syntyi valmistusohje. Sen jälkeen käsitellään putkistoista tehtyjä dokumentaatioita, joita oli jo valmistettu lainsäädännön vaihtumisen jälkeen. Kokoamalla em. tutkimukset on saatu aikaiseksi ohjeistus, jota seuraamallavalmistaja voi valmistaa putkiston, joka täyttää lain ja asetuksen vaatimuksen.Tässä työssä ei tarkasteltu tuotannon tehostamista asennusolosuhteissa eikä laatutavoitteita. Tuloksena syntyi ohjeistus, joka huomioi erilaiset lait ja asetukset, kun kyseessä on putkistojen hankinta, asennus tai korjaus putkistoissa, joita viranomaismääräykset ja asetukset valvovat. Putkistojen asennuksen ja tuotannon tehostaminen sekä automatisoinnin tai mekanisoinnin käyttö hitsauksessa ja hitsauksen laadun varmistus voivat olla tämän diplomityön jälkeisiä tutkimusaiheita.

Genome-wide prediction of matrix attachment regions that increase gene expression in mammalian cells.

Gene transfer in eukaryotic cells and organisms suffers from epigenetic effects that result in low or unstable transgene expression and high clonal variability. Use of epigenetic regulators such as matrix attachment regions (MARs) is a promising approach to alleviate such unwanted effects. Dissection of a known MAR allowed the identification of sequence motifs that mediate elevated transgene expression. Bioinformatics analysis implied that these motifs adopt a curved DNA structure that positions nucleosomes and binds specific transcription factors. From these observations, we computed putative MARs from the human genome. Cloning of several predicted MARs indicated that they are much more potent than the previously known element, boosting the expression of recombinant proteins from cultured cells as well as mediating high and sustained expression in mice. Thus we computationally identified potent epigenetic regulators, opening new strategies toward high and stable transgene expression for research, therapeutic production or gene-based therapies.

Sairaalan tarkkailuosaston hyödyt ja kustannukset

Työn tavoitteena oli tutkia Etelä-Karjalan keskussairaalaan mahdollisesti perustettavan yhdistelmätarkkailuosaston kannattavuutta määrittämällä osaston kustannukset ja hyödyt. Määritettyjen kustannusten avulla vertailtiin päivystys-potilaiden eri hoitoketjuvaihtoehtoja. Lähdeaineistona työssä käytettiin sairaalan omia materiaaleja ja tilastoja, henkilöstön haastatteluja sekä muita kirjallisuuslähteitä. Kirjallisuuden mukaan sairaalan ylikuormitus johtuu sen huonosta potilasvirtojen kulusta, jota voidaan parantaa joko toimintaa tehostamalla tai investoimalla uusiin resursseihin. Eräs vaihtoehto ylikuormituksen purkuun on tarkkailuosaston perustaminen, joka ei ole vain lisätila vaan potilasvirtojen ohjauskeino. Tehtyjen laskemien mukaan Etelä-Karjalan keskussairaalan tarkkailuosasto vaikuttaisi kannattavalta. Tarkkailuosaston kustannukset vuodessa olisivat 1 099 745 €, tuotot 1 170 108 € ja voitto 70 363 €. Osaston perustamiskustannukset olisivat 950 324 €. Hoitoketjujen potilaskohtaisen kannattavuuden mukaan päivystyspotilaiden tarkkailu ja lyhytaikainen hoito kannattaisi tehdä joko vuodeosastoilla tai tarkkailuosastolla, mutta ei tappiota tuottavalla ensiapupoliklinikalla. Tarkkailuosastosta olisi taloudellisten tekijöiden lisäksi myös muita hyötyjä ja haittoja sairaalan sidosryhmille. Sairaalan valitsemasta ylikuormituksen vähentämiskeinosta riippumatta, on sen tärkeätä aloittaa jo lähivuosina valmistautuminen väestön ikääntymisen aiheuttamaan lisääntyvään kuormitukseen.

Safety and Immunogenicity of a Recombinant Adenovirus Serotype 35-Vectored HIV-1 Vaccine in Adenovirus Serotype 5 Seronegative and Seropositive Individuals.

BACKGROUND: Recombinant adenovirus serotype 5 (rAd5)-vectored HIV-1 vaccines have not prevented HIV-1 infection or disease and pre-existing Ad5 neutralizing antibodies may limit the clinical utility of Ad5 vectors globally. Using a rare Ad serotype vector, such as Ad35, may circumvent these issues, but there are few data on the safety and immunogenicity of rAd35 directly compared to rAd5 following human vaccination. METHODS: HVTN 077 randomized 192 healthy, HIV-uninfected participants into one of four HIV-1 vaccine/placebo groups: rAd35/rAd5, DNA/rAd5, and DNA/rAd35 in Ad5-seronegative persons; and DNA/rAd35 in Ad5-seropositive persons. All vaccines encoded the HIV-1 EnvA antigen. Antibody and T-cell responses were measured 4 weeks post boost immunization. RESULTS: All vaccines were generally well tolerated and similarly immunogenic. As compared to rAd5, rAd35 was equally potent in boosting HIV-1-specific humoral and cellular immunity and responses were not significantly attenuated in those with baseline Ad5 seropositivity. Like DNA, rAd35 efficiently primed rAd5 boosting. All vaccine regimens tested elicited cross-clade antibody responses, including Env V1/V2-specific IgG responses. CONCLUSIONS: Vaccine antigen delivery by rAd35 is well-tolerated and immunogenic as a prime to rAd5 immunization and as a boost to prior DNA immunization with the homologous insert. Further development of rAd35-vectored prime-boost vaccine regimens is warranted.

One-sided jet at milliarcsecond scales in LS I +61˚ 303

We present Very Long Baseline Interferometry (VLBI) observations of the high mass X-ray binary LS I +61˚303, carried out with the European VLBI Network (EVN). Over the 11 hour observing run, performed ~10 days after a radio outburst, the radio source showed a constant flux density, which allowed sensitive imaging of the emission distribution. The structure in the map shows a clear extension to the southeast. Comparing our data with previous VLBI observations we interpret the extension as a collimated radio jet as found in several other X-ray binaries. Assuming that the structure is the result of an expansion that started at the onset of the outburst, we derive an apparent expansion velocity of 0:003 c, which, in the context of Doppler boosting, corresponds to an intrinsic velocity of at least 0:4 c for an ejection close to the line of sight. From the apparent velocity in all available epochs we are able to establish variations in the ejection angle which imply a precessing accretion disk. Finally we point out that LS I +61˚303, like SS 433 and Cygnus X-1, shows evidence for an emission region almostorthogonal to the relativistic jet