Cutting Edge: The Heat Shock Protein gp96 Activates Inflammasome-Signaling Platforms in APCs

Several heat shock proteins (HSPs) prime immune responses, which are, in part, a result of activation of APCs. APCs respond to these immunogenic HSPs by upregulating costimulatory molecules and secreting cytokines, including IL-1beta. These HSP-mediated responses are central mediators in pathological conditions ranging from cancer, sterile inflammation associated with trauma, and rheumatoid arthritis. We tested in this study the requirement of inflammasomes in the release of IL-1beta by one immunogenic HSP, gp96. Our results show that murine APCs activate NLRP3 inflammasomes in response to gp96 by K(+) efflux. This is shown to initiate inflammatory conditions in vivo in the absence of additional known inflammasome activators or infection. These results document a novel mechanism by which proteins of endogenous origin, the HSPs, can modulate an inflammatory response following their release from aberrant cells

Enhancing T Cell and NK Cell Mediated anti-Tumor Immune Responses Through the use of Immunotherapy

Thesis (Ph.D.)--University of Rochester. School of Medicine & Dentistry. Dept. of Microbiology and Immunology, 2013.Cancer immunotherapy is a rapidly expanding field that has the potential to control not only primary tumor growth but also the growth of distant metastases. In particular, tumor metastasis to the peritoneal cavity is immunologically intriguing. Upon release into the peritoneal cavity, tumor cells initially attach to the omentum, a tissue consisting of organized immune aggregates embedded in adipose tissue. Indeed, it is to these immune aggregates that tumor cells initially bind and thrive. Despite the proximity of tumor cells and immune cells, a de novo productive immune response does not occur on the omentum. Thus, we explored various mechanisms by which to generate an immune response on the omentum. Intraperitoneal injection of particulate antigen resulted in its localization to the omentum, but only minimal cytokine and cellular responses. Immunization with lethally irradiated tumor cells however, was able to induce a more robust response, namely the expansion of potentially tumor-specific T cells. Subsequently, we challenged mice that had been immunized with lethally irradiated Colon38, a murine colon tumor cell line, to evaluate the quality of the induced anti-tumor response. Following immunization, mice demonstrated both local and peripheral immunity to specific tumor challenge, which required T cells. Interestingly, mice immunized with Colon38 also demonstrated potent non-specific anti-tumor immunity when challenged with an unrelated syngeneic tumor cell line in the peritoneal cavity, which was dependent on NK cells. Since T and NK cells are important in mediating anti-tumor immunity, we explored if IL-2, a potent stimulator of both populations, could alter tumor growth in a setting not involving pre-immunization. Mice were injected with two different clones of B16 cells that expressed different levels of IL-2. Lower levels of IL-2 were able to increase T and NK cell frequencies in tumors, but the alteration in tumor growth induced by this level of IL-2 was modest. Higher levels of IL-2 were able to prevent tumors by inducing dormancy, which was dependent on both T and NK cells. These studies highlight the potent anti-tumor capabilities of T and NK cells in both preventing and treating tumor growth and suggest new treatment strategies for malignant disease

Phenotypically distinct helper NK cells are required for gp96-mediated anti-tumor immunity

A working group, which was established within the Network of Repository Managers (RepManNet), has dealt with common certifications for repositories. In addition, current requirements of the research funding agencies FWF and EU were also taken into account. The Core Trust Seal was examined in more detail. For this purpose, a questionnaire was sent to those organizations that are already certified with CTS in Austria. The answers were summarized and evaluated anonymously. It is recommended to go for a repository certification. Moreover, the development of a DINI certificate in Austria is strongly suggested.Eine Arbeitsgruppe, die im Rahmen des Netzwerks für RepositorienmanagerInnen (RepManNet) entstanden ist, hat sich mit gängigen Zertifizierungen für Repositorien beschäftigt. Weiters wurden aktuelle Vorgaben der Forschungsförderer FWF und EU herangezogen. Das Core Trust Seal wurde genauer betrachtet. Hierfür wurden jenen Organisationen, die in Österreich bereits mit CTS zertifiziert sind, ein Fragebogen übermittelt. Die Antworten wurden anonymisiert zusammengefasst und ausgewertet. Plädiert wird für eine Zertifizierung von Repositorien und die Entwicklung einer DINI-Zertifizierung in Österreich

Extensive identification of genes involved in congenital and structural heart disorders and cardiomyopathy

Clinical presentation of congenital heart disease is heterogeneous, making identification of the disease-causing genes and their genetic pathways and mechanisms of action challenging. By using in vivo electrocardiography, transthoracic echocardiography and microcomputed tomography imaging to screen 3,894 single-gene-null mouse lines for structural and functional cardiac abnormalities, here we identify 705 lines with cardiac arrhythmia, myocardial hypertrophy and/or ventricular dilation. Among these 705 genes, 486 have not been previously associated with cardiac dysfunction in humans, and some of them represent variants of unknown relevance (VUR). Mice with mutations in Casz1, Dnajc18, Pde4dip, Rnf38 or Tmem161b genes show developmental cardiac structural abnormalities, with their human orthologs being categorized as VUR. Using UK Biobank data, we validate the importance of the DNAJC18 gene for cardiac homeostasis by showing that its loss of function is associated with altered left ventricular systolic function. Our results identify hundreds of previously unappreciated genes with potential function in congenital heart disease and suggest causal function of five VUR in congenital heart disease