This protocol describes methods to identify, purify and culture CD1d restricted invariant natural killer T (NKT) cells from mouse tissue or human blood samples. The methods for identification and purification ofNKT cells are based on the interaction betweenNKT cell receptor and its ligand. TheNKT ...
By transducing CAR structure into the NKT cell, the CAR-NKT cell has been created as a novel genetically-engineered cell with different abilities that can affect the cancer cells differently. Thevarious actions of these cells against tumors have been demonstrated in Fig.2. The CAR-NKT cell can...
to each well, and after an additional 16 h of culture, cells were collected with a cell harvester and uptake of radio- activity was measured with a Betaplate reader (Wallac, Gaithersburg, USA). Cytokine secretion following in vivo α-GalCer treat- ...
To examine whether our treatment protocol would change the composition of intestinal flora, we applied the DNA microarray system referred to as ‘FloraArray’28 and made a comprehensive analysis for intestinal flora derived from KCV-treated mice and control mice. To compare the signal intensities ...
Cell culture. HEK 293T, Hepa 1–6 (mouse hepatocyte cell line), Jurkat (human T-cell line) and B16 mouse melanoma cells were cultured in DMEM supplemented with 10% FBS. Mouse NKT hybridoma cells DN32.D3 were obtained from Dr Bendelac (University of Chicago) and were maintained in RPMI...
(A–E) Sorted splenic NKT cells from C57BL/6 mice were labeled with 5 μM CellTrace Violet (CTV) and stimulated for 3 days in the indicated culture conditions. (A) The data show percentages of cell survival as measured by live/dead marker staining (left panel) and cell proliferation (...
A.H. wrote the clinical protocol and was the principal investigator of the clinical trial. G.D., L.S.M., A.H., X.X. and G.T. developed the CAR construct selected for the trial and tested it in NKT cells. A.H., A.C.S. and B.D.W. performed pre-therapy and post-therapy ...
development of autoimmune diseases in certain models [16,17]. Using a similar protocol, we can efficiently produce, from the total splenic CD8+T cell population, large numbers of CD8+T cells that co-express various NK markers. These cells are therefore referred to as CD8+NKT-like cells. We...
Positive controls consisted of splenocytes processed at the same time under the same protocol. Negative controls consisted of cells stained with fluorescein isothiocyanate, phycoerythrin, or APC-isotype antibodies (BD Pharmingen). Cells were resuspended in violet dye (live/dead cell fixable staining,...
1. A microparticle or nanoparticle artificial antigen presenting cell for stimulation and expansion of NKT cells, comprising: (a) a paramagnetic iron dextran bead; and, on the surface of the bead, (b) a population of CD1d antigen presenting complexes that activate T cell receptors (TCRs) of ...