A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes

A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. The Huntington's Disease Collaborative Research Group.

Dendritic cells (DCs) are antigen-presenting cells with a unique ability to induce primary immune responses. DCs capture and transfer information from the outside world to the cells of the adaptive immune system. DCs are not only critical for the induction of primary immune responses, but may also be important for the induction of immunological tolerance, as well as for the regulation of the type of T cell-mediated immune response. Although our understanding of DC biology is still in its infancy, we are now beginning to use DC-based immunotherapy protocols to elicit immunity against cancer and infectious diseases.

Immunobiology of Dendritic Cells

A tandem repeat in DNA is two or more contiguous, approximate copies of a pattern of nucleotides. Tandem repeats have been shown to cause human disease, may play a variety of regulatory and evolutionary roles and are important laboratory and analytic tools. Extensive knowledge about pattern size, copy number, mutational history, etc. for tandem repeats has been limited by the inability to easily detect them in genomic sequence data. In this paper, we present a new algorithm for finding tandem repeats which works without the need to specify either the pattern or pattern size. We model tandem repeats by percent identity and frequency of indels between adjacent pattern copies and use statistically based recognition criteria. We demonstrate the algorithm’s speed and its ability to detect tandem repeats that have undergone extensive mutational change by analyzing four sequences: the human frataxin gene, the human β T cell receptor locus sequence and two yeast chromosomes. These sequences range in size from 3 kb up to 700 kb. A World Wide Web server interface at c3.biomath.mssm.edu/trf.html has been established for automated use of the program.

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Tandem repeats finder: a program to analyze DNA sequences

Signal transduction is initiated by complex protein-protein interactions between ligands, receptors and kinases, to name only a few. It is now becoming clear that lipid micro-environments on the cell surface -- known as lipid rafts -- also take part in this process. Lipid rafts containing a given set of proteins can change their size and composition in response to intra- or extracellular stimuli. This favours specific protein-protein interactions, resulting in the activation of signalling cascades.

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Lipid rafts and signal transduction

Introduction. Remarkable progress has been made in the treatment of relapsed/refractory Acute Lymphocytic Leukemia and Non-Hodgkin Lymphoma with CAR-CD19 T cells. In contrast, progress against CD19-negative hematological cancers and solid tumors has been limited. Intensive efforts to optimize cellular therapeutics for better efficacy include provision of cytokine support and countering immunosuppression. However, lack of sufficient antigen is a significant additional hurdle that CAR-T therapeutics for solid tumors must overcome. We present a novel strategy to utilize CD19 for sustained antigen presentation in order to promote cellular therapeutic expansion, efficacy and persistence. The strategy, called IMPACT (Integrated Modules oPtimize Adoptive Cell Therapy), employs a methodology that is modular in design and and can be applied to diverse antigens and tumor types, yet retains the well-established advantages of CAR T cells directed to CD19. Experimental Procedures. A 3rd generation CAR-T was constructed in a lentiviral vector using the FMC63 anti-CD19 scFv for antigen recognition. A second expression cassette, cloned downstream of the CAR-T cassette, was constructed using a secretion signal sequence and exons 1 - 4 of the human CD19 protein linked in frame to a trastuzumab-derived Vh/Vl sequence. This integrated-gene module thus encodes a fusion protein (FP): CD19-anti-HER2-scFv. Results. ELISA and flow cytometry analyses were used to demonstrate bridging between HER2+ cells and the FMC63 mAb via the CD19 domain of the FP. ELISA analyses further demonstrated wild type binding of the 2 FP domains (i.e. CD19/FMC63 and the scFv/HER2). In vitro cytotoxicity studies showed robust redirected killing of HER2+ (CD19-) tumor cells by CAR19 T cells that is mediated by the secreted FP. The efficiency of cytotoxicity was enhanced by the presence of CD19+ B cells. A tumor xenograft model was used to demonstrate effective killing in vivo when HER2+ BT474 cells were injected in one flank and CD19+ RAMOS cell were injected in the opposite flank, but not when HER2+ cell were injected alone. Thus, in vivo, redirected killing required CD19+ cells for expansion and persistence sufficient to promote anti-HER2 cytotoxicity. Conclusions. A CD19-anti-HER2-scFv FP was able to redirect the cytotoxicity of CAR19 T cells in order to kill HER2+ tumor cells. In vivo modeling demonstrated that CAR-T cell expansion by CD19 recognition enabled efficient HER2+ cell cytotoxicity. The IMPACT technology is modular, allowing for FPs having diverse protein domains paired with diverse, distinct scFvs to allow CAR T cells to CD19 or other targets to be used to kill any tumor. We have further developed the technology to address antigens of utility for hematopoietic and solid tumors, including BCMA, ROR1, and PD-L1. The technology is broadly applicable and synergistic with existing cellular therapies. Citation Format: Christine Ambrose, Lihe Su, Lan Wu, Roy R. Lobb, Paul D. Rennert. CAR T cells specific for CD19 can be redirected to kill CD19 negative tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3768. doi:10.1158/1538-7445.AM2017-3768

Abstract 3768: CAR T cells specific for CD19 can be redirected to kill CD19 negative tumors

BAFF selectively enhances the survival of plasmablasts generated from human memory B cells (vol 112, pg 286, 2003)

The disclosure provides a non-naturally occurring BAFF-R glycoprotein having a deletion in the extracellular domain which results in an altered O-linked glycosylation pattern. The disclosure also provides methods and pharmaceutical compositions for treating B-cell- and T-cell-mediated disorders.

Method of treating B cell cancers by administering truncated BAFF receptors

CAR19 T Cells Redirected to Novel Antigens Mediate Robust Cytotoxicity Against Diverse Malignancies

e15046Background: CAR T cells targeting CD19 (CAR19s) can eradicate B cell leukemias and lymphomas. We have engineered CAR19s to secrete a wide variety of retargeting fusion proteins (FP) by encodi...

Christine Ambrose

Papers

Abstract 3768: CAR T cells specific for CD19 can be redirected to kill CD19 negative tumors

BAFF selectively enhances the survival of plasmablasts generated from human memory B cells (vol 112, pg 286, 2003)

Method of treating B cell cancers by administering truncated BAFF receptors

CAR19 T Cells Redirected to Novel Antigens Mediate Robust Cytotoxicity Against Diverse Malignancies

Activity of CAR19 T cells secreting impact fusion proteins against hematopoietic and solid tumors.