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Autoimmune diseases and in particular type 1 diabetes rely heavily on treatments that target the symptoms rather than prevent the underlying disease. One of the barriers to better therapeutic strategies is the inability to detect and efficiently target rare autoreactive T-cell populations that are major drivers of these conditions. Here, we develop a unique artificial antigen-presenting cell (aAPC) system from biocompatible polymer particles that allows specific encapsulation of bioactive ingredients. Using our aAPC, we demonstrate that we are able to detect rare autoreactive CD4 populations in human patients, and using mouse models, we demonstrate that our particles are able to induce desensitization in the autoreactive population. This system provides a promising tool that can be used in the prevention of autoimmunity before disease onset.

Autoimmune diseases and in particular type 1 diabetes rely heavily on treatments that target the symptoms rather than prevent the underlying disease. One of the barriers to better therapeutic strategies is the inability to detect and efficiently target rare autoreactive T-cell populations that are major drivers of these conditions. Here, we develop a unique artificial antigen presenting cell (aAPC) system from biocompatible polymer particles that allows specific encapsulation of bioactive ingredients. Using our aAPC we demonstrate that we are able to detect rare autoreactive CD4 populations in human patients and using mouse models we demonstrate that our particles are able to induce desensitization in the autoreactive population. This system provides a promising tool that can be used in the prevention of autoimmunity before disease onset.

BackgroundChimeric antigen receptor (CAR) T cell therapy has shown outstanding benefit in hematological malignancies with three autologous CAR-T therapies commercially available and several in clinical development. The use of autologous T cells to manufacture CAR therapies has several disadvantages including production time, cost, lengthy time to treatment and dependence on patient T cell functionality. An allogeneic off the shelf CAR product can overcome these issues. We use invariant natural killer T (iNKT) cells as basis for our allogeneic cell therapy platform. Invariant NKT cells are innate-like lymphocytes that bridge innate and adaptative immune response to promote anti-cancer immunity. iNKT cells share characteristics of T cells and Natural Killer (NK) cells, expressing both an invariant T Cell Receptor (iTCR) and canonical NK receptors. They can be activated by recognition of lipid antigens through the iTCR, pro-inflammatory cytokines and recognition of stress ligands. Moreover, iNKT do not cause Graft versus Host Disease making them an ideal platform for allogeneic CAR cell therapy. Here we describe a novel allogeneic iNKT-CAR product targeting BCMA designed to promote effective anti-cancer immunityMethodsOur proprietary CARDIS TM platform is a 2-stage discovery process where screening of highly diverse scFv libraries via phage display is followed by selection of potent CARs using mammalian display platform. Using this high throughput approach, we can interrogate large cell-based CAR libraries for specific binding and activation simultaneously, as well as eliminate tonic signaling at an early stage. This novel platform enabled rapid identification of a candidate CAR (agenT-F6) for the targeting of BCMA. Anti-cancer efficacy of stably expressed BCMA CAR in expanded iNKT cells has been testedResultsAgent-F6 expressed at high levels in iNKT cells. Functional characterization demonstrated potent cytotoxic activity of agent-F6 expressing iNKT cells against human hematologic tumor cell lines expressing BCMA in vitro. Activated agenT-F6 iNKT cells display a pro-inflammatory phenotype when challenged with tumor cell lines. Furthermore, using a multiple myeloma in vivo xenograft model, infusion of agenT-F6 iNKT cells showed comparable tumor control to a clinical-stage BCMA CAR reference.ConclusionsMiNK therapeutics is developing the next generation allogeneic CAR-iNKT therapies by using 1) a CAR discovery platform that allows more rapid and efficient identification of antigen-specific and biologically potent CAR candidates and 2) an iNKT cell platform naturally lacking alloreactivity that allows rapid engineering and expansion of an off-the-shelf CAR product.Ethics ApprovalAll procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards

Legionella pneumophila has been shown to utilize the icm/dot type IV secretion system for pathogenesis. This system was shown to be composed of icm / dot complex components and accessory proteins, as well as a large number of translocated substrates. Bioinformatic analysis of the regulatory regions of all the genes revealed that several icm / dot genes, as well as two genes encoding icm / dot translocated substrates, contain the conserved CpxR regulatory element, a regulator that has been shown previously to control the expression of the icmR gene. An experimental analysis, which included a comparison of gene expression in a L. pneumophila wild-type strain and gene expression in a cpxR deletion mutant, construction of mutants with mutations in the CpxR conserved regulatory elements, controlled expression studies, and mobility shift assays, demonstrated the direct relationship between the CpxR regulator and the expression of the genes. Furthermore, genomic analysis identified nine additional genes that contain a putative CpxR regulatory element; five of these genes (two legA genes and three ceg genes) were suggested previously to be putative icm / dot translocated substrates. The three ceg genes identified, which were shown previously to contain a putative PmrA regulatory element, were found here to be regulated by both CpxR and PmrA. The other six genes (two legA genes and four new genes products were found to be regulated by CpxR. Moreover, using the CyaA translocation assay, these nine gene products were found to be translocated into host cells in an Icm/Dot-dependent manner. Our results establish that the CpxR regulator is a fundamental regulator of the icm / dot type IV secretion system in L. pneumophila .