Consensus guidelines for the definition, detection and interpretation of immunogenic cell death.

Cells succumbing to stress via regulated cell death (RCD) can initiate an adaptive immune response associated with immunological memory, provided they display sufficient antigenicity and adjuvanticity. Moreover, multiple intracellular and microenvironmental features determine the propensity of RCD to drive adaptive immunity. Here, we provide an updated operational definition of immunogenic cell death (ICD), discuss the key factors that dictate the ability of dying cells to drive an adaptive immune response, summarize experimental assays that are currently available for the assessment of ICD in vitro and in vivo, and formulate guidelines for their interpretation

Combined Tumor Cell-Based Vaccination and Interleukin-12 Gene Therapy Polarizes the Tumor Microenvironment in Mice

Tumor progression depends on tumor milieu, which influences neovasculature formation and immunosuppression. Combining immunotherapy with antiangiogenic/antivascular therapy might be an effective therapeutic approach. The aim of our study was to elaborate an anticancer therapeutic strategy based on the induction of immune response which leads to polarization of tumor milieu. To achieve this, we developed a tumor cell-based vaccine. CAMEL peptide was used as a B16-F10 cell death-inducing agent. The lysates were used as a vaccine to immunize mice bearing B16-F10 melanoma tumors. To further improve the therapeutic effect of the vaccine, we combined it with interleukin (IL)-12 gene therapy. IL-12, a cytokine with antiangiogenic properties, activates nonspecific and specific immune responses. We observed that combined therapy is significantly more effective (as compared with monotherapies) in inhibiting tumor growth. Furthermore, the tested combination polarizes the tumor microenvironment, which results in a switch from a proangiogenic/immunosuppressive to an antiangiogenic/immunostimulatory one. The switch manifests itself as a decreased number of tumor blood vessels, increased levels of tumor-infiltrating CD4+, CD8+ and NK cells, as well as lower level of suppressor lymphocytes (Treg). Our results suggest that polarizing tumor milieu by such combined therapy does inhibit tumor growth and seems to be a promising therapeutic strategy

Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

Intrinsic nanocrystalline diamond (NCD) films have been proven to be promising substrates for the adhesion, growth and osteogenic differentiation of bone-derived cells. To understand the role of various degrees of doping (semiconducting to metallic-like), the NCD films were deposited on silicon substrates by a microwave plasma-enhanced CVD process and their boron doping was achieved by adding trimethylboron to the CH4:H2 gas mixture, the B∶C ratio was 133, 1000 and 6700 ppm. The room temperature electrical resistivity of the films decreased from >10 MΩ (undoped films) to 55 kΩ, 0.6 kΩ, and 0.3 kΩ (doped films with 133, 1000 and 6700 ppm of B, respectively). The increase in the number of human osteoblast-like MG 63 cells in 7-day-old cultures on NCD films was most apparent on the NCD films doped with 133 and 1000 ppm of B (153,000±14,000 and 152,000±10,000 cells/cm2, respectively, compared to 113,000±10,000 cells/cm2 on undoped NCD films). As measured by ELISA per mg of total protein, the cells on NCD with 133 and 1000 ppm of B also contained the highest concentrations of collagen I and alkaline phosphatase, respectively. On the NCD films with 6700 ppm of B, the cells contained the highest concentration of focal adhesion protein vinculin, and the highest amount of collagen I was adsorbed. The concentration of osteocalcin also increased with increasing level of B doping. The cell viability on all tested NCD films was almost 100%. Measurements of the concentration of ICAM-1, i.e. an immunoglobuline adhesion molecule binding inflammatory cells, suggested that the cells on the NCD films did not undergo significant immune activation. Thus, the potential of NCD films for bone tissue regeneration can be further enhanced and tailored by B doping and that B doping up to metallic-like levels is not detrimental for cells

Handling stress as a measurement of personality in great tit nestlings (Parus major)

Interest in personality is growing in a wide range of disciplines, but only in a few systems it is possible to assess the survival value of personality. Field studies looking at the relationship between personality and survival value early in life are greatly hampered by the fact that personality can at present only be assessed after individuals become independent from their parents. In passerines, for example, this is often after a period of intensive selection for the survival on fledglings. The main aim of this study is therefore to develop a method to measure personality before this period of selection. For this purpose, we developed the handling stress (HS) test. We measured HS in 14-d-old great tit nestlings by counting the number of breast movements (breath rate) in four subsequent 15-s bouts for 1 min; before and after they were socially isolated from their siblings for 15 min. To calculate the repeatability of HS, we repeated the test 6 mo later. To assess the relat! ionship between HS and exploratory behaviour, we correlated the outcome of both tests. We ran tests both on birds of lines selected for extreme personality and on wild birds from a natural population. We found that birds selected for fast exploration reacted more to HS compared with birds selected for slow exploration and that HS was repeatable in different life phases. We confirmed this by finding an increase in the HS with increasing exploratory scores in wild birds. These results show that we can use the HS test as a measurement of personality, making it a potential tool for studying the relationship between personality and survival value early in life.

Social personality traits in chimpanzees: temporal stability and structure of behaviourally assessed personality traits in three captive populations

Animals of many species show consistency in behaviour across time and contexts that differs from other individuals' behaviour in the same population. Such ‘personality’ affects fitness and has therefore become an increasingly relevant research topic in biology. However, consistent variation in social behaviour is understudied. In socially living species, behaviour occurs in a social environment and social interactions have a significant influence on individual fitness. This study addressed personality in social behaviour of 75 captive chimpanzees in three zoos by coding observed behaviour. Fifteen behavioural variables were significantly repeatable (range 0.21–0.93) in at least two of the three zoos. The behaviours showed considerable long-term stability across 3 years, which did not differ from the short-term repeatability. The repeatable behaviours were then analysed with factor analyses. They formed five independent factors, three of which consisted of social traits and were labelled ‘sociability’, ‘positive affect’ and ‘equitability’. The two non-social behaviour factors were labelled ‘anxiety’ and ‘activity’. The factor scores were analysed for sex and population differences. Males had higher factor scores in all traits except ‘sociability’. The factor scores differed also between the zoos, implying considerable external effects in trait expression. The results show that chimpanzees show personality in a broad range of social and non-social behaviours. The study highlights the importance of assessing personality in the social behaviour, especially in cohesive social species, as only then can we understand the consequences of personality in socially living species