31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Distribucción de Calanus spp. determinada utilizando un sistema de identificación genética

The morphological similarity of Calanus species necessitates that the only unambiguous way of discriminating between the different species at any developmental stage is with molecular tools. We have developed a simple molecular technique to distinguish between the four species of Calanus copepods found in the North Atlantic (Calanus helgolandicus, C. finmarchicus, C. glacialis and C. hyperboreus) at any life stage. This system involves the PCR amplification of a region of the 16S rRNA gene, followed by Restriction Fragment Length Polymorphism (RFLP) analysis of the amplified product. This paper describes the application of the technique to a number of Calanus samples taken from a wide geographical range within the North Atlantic. Samples were acquired from sites ranging in latitude from the English Channel (50&#176;N) to Troms&oslash; (69&#176;N), including many regions of co-occurrence. A comparison has been made between the distribution of Calanus as determined by molecular techniques and distributions established using traditional morphological identification. The molecular analysis has clearly shown extended areas of distribution and co-occurrence and has confirmed the intraspecific conservation of the restriction sites and therefore the reliability of the technique.La semejanza morfológica de las especies de Calanus obliga a que la única manera clara de discriminar entre las distintas especies en cualquier estadio de desarrollo sea mediante herramientas moleculares. Hemos desarrollado una técnica molecular simple para distinguir las cuatro especies de copépodos del género Calanus que se encuentran en el Atlántico Norte (Calanus helgolandicus, C. finmarchicus, C. glaciales and C. hyperboreus), sea cual sea el estadio de su ciclo biológico. Este sistema implica la amplificación mediante PCR de una región del gen 16S rRNA, seguida de análisis de Polimorfismo de Longitud de Fragmentos de Restricción de (RFLP) del producto amplificado. Este trabajo describe la aplicación de la técnica a varias muestras de Calanus procedentes de una amplia región geográfica en el Atlántico Norte. Se obtuvieron muestras de localidades que iban desde el canal de la Mancha (50°N) a Tromsø (69°N), incluyendo muchas regiones de presencia conjunta. Se ha hecho una comparación entre la distribución de Calanus, determinada por las técnicas moleculares, y las distribuciones sobre la base de la identificación morfológica tradicional. El análisis molecular ha demostrado claramente áreas extendidas de distribución y ocurrencia conjunta y ha confirmado la conservación intraspecífica de los lugares de restricción, con lo que se ha confirmado la fiabilidad de la técnica