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To carry out tumour recognition and fine MHC restriction analysis, we isolated T-cell clones from the EGFR875 889 reactive CD4 T-cell lines of the four normal blood donors.
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For RNA isolation isolated T cells were pelleted, immediately placed in RNA Later (Applied Biosystems/Ambion, Austin, TX) and after overnight infiltration at 4°C they were kept in RNA Later at −70°C until the isolation of total RNA.
To overcome this problem, we have isolated T cell receptor (TCR) genes from four different HPV16E7-specific healthy donor and patient-derived human cytotoxic T lymphocyte (CTL) clones.
Immunologist David Hafler of Brigham and Women's Hospital and Harvard Medical School in Boston, Massachusetts, and his colleagues isolated T cells from the pancreatic lymph nodes of diabetic and nondiabetic patients--something that hadn't been done before.
We investigated the cellular interactions that occur after adoptive transfer of isolated T cell populations into T cell deficient hosts.
Microparticles (MP) generated by stimulated T cells (MPT) display similar monocyte activating ability to whole T cells, isolated T cell membranes, or solubilized T cell membranes.
The resulting average area of an isolated T cell comprised 51 µm2 (range being 28 80 µm2; standard deviation 10.65 µm2).
Freshly isolated T cell-depleted splenocytes were CFSE-labeled and the fate of the transferred cells determined by flow cytometry and immunofluorescence microscopy.
The differing proliferation results in PBMC cultures as compared to isolated T cell cultures led us to investigate the production of sCD25 as a function of PBMC proliferation (Figure 7).
While isolated cells (28 to 80 µm2) are counted directly, the number of cells contained in a conglomerate is estimated by dividing the area of the conglomerate in thin tissues sections (≤6 µm) by the median area covered by an isolated T cell which we determined as 58 µm2.
Freshly isolated T cells from WT and Olig1−/− MOG-immunized mice were stimulated with different concentrations of MOG.
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