We developed an innovative and efficient, feeder-free culture method to genetically modify and expand peripheral blood-derived NK cells with high proliferative capacity, while preserving the responsiveness of their native activating receptors. Activated peripheral blood NK cells were efficiently transduced by a retroviral vector, carrying a second-generation CAR targeting CD19. CAR expression was demonstrated across the different NK-cell subsets. CAR.CD19-NK cells display higher antileukemic activity toward CD19+ cell lines and primary blasts obtained from patients with B-cell precursor ALL compared with unmodified NK cells. In vivo animal model data showed that the antileukemia activity of CAR.CD19-NK cell is superimposable to that of CAR-T cells, with a lower xenograft toxicity profile. These data support the feasibility of generating feeder-free expanded, genetically modified peripheral blood NK cells for effective "off-the-shelf" immuno-gene-therapy, while their innate alloreactivity can be safely harnessed to potentiate allogeneic cell therapy.
Antigens, CD19/immunology , Cell- and Tissue-Based Therapy/methods , Immunotherapy, Adoptive/methods , Killer Cells, Natural/transplantation , Leukocytes, Mononuclear/immunology , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/therapy , Receptors, Chimeric Antigen/immunology , Animals , Apoptosis , Cell Proliferation , Cytotoxicity, Immunologic/immunology , Humans , Killer Cells, Natural/immunology , Mice , Mice, Inbred NOD , Mice, SCID , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/immunology , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/pathology , Tumor Cells, Cultured , Xenograft Model Antitumor Assays
Recent experimental findings have shown the ability of mesenchymal stromal cells (MSCs) to home to damaged tissues and to produce paracrine factors with anti-inflammatory properties, potentially resulting in reduction of inflammation and functional recovery of the damaged tissues. Prompted by these intriguing properties and on the basis of encouraging preclinical data, MSCs are currently being studied in several immune-mediated disorders. Inflammatory bowel diseases (IBD) represent a setting in which MSCs-based therapy has been extensively investigated. Phase I and II studies have documented the safety and feasibility of MSCs. However, efficacy results have so far been conflicting. In this review, we will discuss the biologic rationale that makes MSCs a promising therapeutic tool for IBD, and analyze recent experimental and clinical findings, highlighting current limitations and future perspectives of MSCs-related immunotherapy for IBD.
Cell Movement/immunology , Inflammatory Bowel Diseases/therapy , Mesenchymal Stem Cell Transplantation/methods , Mesenchymal Stem Cells/immunology , Adaptive Immunity/immunology , Animals , Clinical Trials as Topic , Humans , Immunity, Innate/immunology , Inflammatory Bowel Diseases/immunology , Mesenchymal Stem Cells/cytology , Treatment Outcome
Apoptosis , Cardiac Output, Low/prevention & control , Glutamic Acid/genetics , Lysine/genetics , Myocardium/enzymology , Neovascularization, Physiologic , Proto-Oncogene Proteins c-akt/metabolism , Animals , Blood Pressure , Cardiac Output, Low/chemically induced , Cardiac Output, Low/pathology , Cardiac Output, Low/physiopathology , Coronary Circulation , Gene Expression Regulation , Mice , Myocardium/pathology , Myocytes, Cardiac/pathology , Proto-Oncogene Proteins c-akt/chemistry , Proto-Oncogene Proteins c-akt/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/metabolism
