Molecular epidemiology of the endemic multiresistance plasmid pSI54/04 of Salmonella Infantis in broiler and human population in Hungary

Salmonella Infantis (SI) became endemic in Hungary where the PFGE cluster B, characterized by a large multiresistance (MDR) plasmid emerged among broilers leading to an increased occurrence in humans. We hypothesised that this plasmid (pSI54/04) assisted dissemination of SI. Indeed, Nal-Sul-Tet phenotypes carrying pSI54/04 occurred increasingly between 2011-2013 among SI isolates from broilers and humans. Characterization of pSI54/04 based on genome sequence data of the MDR strain SI54/04 indicated a size of ~277 kb and a high sequence similarity with the megaplasmid pESI of SI predominant in Israel. Molecular characterization of 78 representative broiler and human isolates detected the prototype plasmid pSI54/04 and its variants of together with novel plasmid associations within the emerging cluster B. To test in vitro and in vivo pathogenicity of pSI54/04 we produced plasmidic transconjugant of the plasmid-free pre-emergent strain SI69/94. This parental strain and its transconjugant have been tested on chicken embryo fibroblasts (CEFs) and in orally infected day old chicks. The uptake of pSI54/04 did not increase the pathogenicity of the strain SI69/94 in these systems. Thus, dissemination of SI in poultry could be assisted by antimicrobial resistance rather than by virulence modules of the endemic plasmid pSI54/04 in Hungary

Insight into the molecular characteristics of new multiresistant clones and plasmids in Salmonella Infantis in poultry and man

The lack of the regular monitoring of Salmonella Infantis together with the effort to reduce prevalence of „top five” serovars lead to a dramatic increase of S. Infantis in poultry with reflection in human population. As a background of this study, a clonal change reported previously in Hungarian S. Infantis strains is remarkable (Nógrády et al., 2007). Accordingly, in the early 2000s, older pansensitive isolates of S. Infantis have been replaced by new tetraresistant clones carrying a large plasmid conferring the multiresistance phenotype. Based on the above finding, our aim is to characterize antimicrobial resistance pheno-and genotypes of recently isolated S. Infantis strains, and to provide the first description of the large multiresistance plasmid in a poultry isolate representing the prevalent tetraresistant Hungarian clone. For this purpose ~300 strains of S. Infantis were tested, originating mostly from broilers and from human clinical samples. Strains intended to represent the current status of S. Infantis infection in poultry and human between 2011-2013. The antimicrobial resistance phenotype, pulsotype and the plasmid content were determined, based on which representative strains were selected for resistance genotyping by PCR microarray. One poultry strain (SI 54/04), representative of the tetraresistant, plasmid containing strains was subjected to genome sequencing (Olasz et al., 2015), giving a basis for the characterization of the large multiresistance plasmid. Resistance phenotyping and PFGE analysis has shown a constant circulation of the former major multiresistant clones and patterns within the newly isolated strains of S. Infantis both in poultry and human. Multiresistance phenotypes were associated mostly with the presence of class 1 integrons (intI1)and the gene tetA for tetracycline resistance, being the prime genetic markers for the carriage of the large multiresistance plasmid. However, the coexistence of the tetA and other plasmid related genes for β-lactam and fluoroquinolone resistance such as blaTEM-1, bla CMY-9 or qnr indicate important and divergent plasmid associations in some of the strains. By sequence analysis we provide the characterization of the multiresistance megaplasmid (pSI54/04) of the strain SI 54/04 which can be considered as the first Hungarian reference plasmid of Salmonella Infantis. The backbone of this IncI type plasmid is a mosaic of resistance (nickel-, mercury resistance), and virulence regions (encoding siderophore Yersiniabactin, fimbriae) potentially promoting survival not only in the vertebrate hosts but also in their environment. Support of OTKA K101564 and the János Bolyai stipend of the Hungarian Academy of Sciences to A. Szmolka is acknowledged

A hereditaer nonpolyposus colorectalis carcinoma szindrómás betegek szűrésének és szoros utánkövetésének fontossága egy családfa bemutatása kapcsán | Significance of the monitoring and screening for hereditary nonpolyposis colorectal carcinoma syndrome patients by presenting a case of a family tree

Absztrakt: Bevezetés: A hereditaer nonpolyposus colorectalis carcinomára jellemző mutációk (HNPCC) autoszomális domináns öröklődésmenetet mutatnak. Leginkább vastagbéldaganatok kialakulásáért felelősek. Célkitűzés: A HNPCC-szindrómás betegek szűrésének és követésének fontosságát szeretnénk hangsúlyozni egy igazolt MMR-gén-mutációt hordozó betegünk jelenlegi és 10 évvel korábbi családfájának összehasonlításával. Betegek és módszer: Hazánkban előforduló, HNPCC-re gyanús családok kiszűrése érdekében alapos családi anamnézist veszünk fel. Amennyiben az immunhisztokémiai és mikroszatellitainstabilitás-vizsgálatok HNPCC-szindrómára utalnak, elvégezzük az MMR-gének szekvenálását. Eredmények: Betegünknél egy, a hMSH2-gén 6. exon két bázispárt érintő deletiója (c.969–970delTC) igazolódott. Tízéves utánkövetés során betegünknél és rokonainál újabb, a HNPCC-re jellemző tumorok jelentek meg. Következtetés: A veszélyeztetett családtagok követése során a szekunder prevenció a jól együttműködő betegeknél hatékony volt. Orv Hetil. 2017; 158(30): 1182–1187. | Abstract: Introduction: Hereditary nonpolyposis colorectal carcinoma (HNPCC) is an autosomal dominant disease, which shows familial clustering. Aim: We would like to emphasize the importance of monitoring the HNPCC syndrome patients by presenting a case of a proven MMR gene mutation carrier and her family tree encompassing 10 years. Materials and method: To screen a suspected HNPCC Hungarian family member we are taking thorough family histories. If the diagnosis of HNPCC was further supported by immunohistology and the microsatellite status, sequencing of the MMR genes was carried out. Results: A novel mutation in exon 6 of the hMSH2 gene leading to the deletion of two nucleotide pairs [c.969-970delTC] was detected in our patient. During the 10-year follow-up period of our patient new HNPCC-associated tumors have developed in several family members. Conslusion: Close surveillance of the patient and its family members at risk was effective, although it requires compliance from the subjects. Orv Hetil. 2017; 158(30): 1182–1187

Identification of Novel Pathogenic Sequence Variants of the Mismatch Repair Genes During Screening for Lynch Syndrome in a Single Centre of Eastern Hungary

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Salivary IL-6 mRNA is a Robust Biomarker in Oral Squamous Cell Carcinoma

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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

Determination of the phylogenetic origins of the Árpád Dynasty based on Y chromosome sequencing of Béla the Third

International audienceWe set out to identify the origins of the Árpád Dynasty based on genome sequencing of DNA derived from the skeletal remains of Hungarian King Béla III (1172-1196) and eight additional individuals (six males, two females) originally interred at the Royal Basilica of Székesfehérvár. Y-chromosome analysis established that two individuals, Béla III and HU52 assign to haplogroups R-Z2125 whose distribution centres near South Central Asia with subsidiary expansions in the regions of modern Iran, the Volga Ural region and the Caucasus. Out of a cohort of 4340 individuals from these geographic areas, we acquired whole-genome data from 208 individuals derived for the R-Z2123 haplogroup. From these data we have established that the closest living kin of the Árpád Dynasty are R-SUR51 derived modern day Bashkirs predominantly from the Burzyansky and Abzelilovsky districts of Bashkortostan in the Russian Federation. Our analysis also reveals the existence of SNPs defining a novel Árpád Dynasty specific haplogroup R-ARP. Framed within the context of a high resolution R-Z2123 phylogeny, the ancestry of the first Hungarian royal dynasty traces to the region centering near Northern Afghanistan about 4500 years ago and identifies the Bashkirs as their closest kin, with a separation date between the two populations at the beginning of the first millennium CE