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Altogether, 10 patients with CLL were included in the study. Seven were clinically in very good condition, and three were in poor condition at the time of sampling. The median age of the patients was 72 years and the median follow-up time was 31 months. All patients were treated with the PI3K inhibitor, Ibrutinib. The subtypes of CLL according to the Hans classification were as follows: 4 were type I, 3 were type II and 3 were type III. Two samples were of chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) combined type and one was of follicular lymphoma. The median number of white blood cells (WBC) in the blood for the patients with CLL at diagnosis was 9.3 x 109/L. The median number of platelets in the blood at diagnosis was 209 x 109/L. The patients were classified according to the Binet stage as follows: 4 were in stage A, 2 in stage B, 2 in stage C and 2 in stage D.
Stimulation of T cells is a critical step in the success of an adoptive immunotherapy (AIT) strategy. Studies have shown that strong co-stimulation can elicit a powerful response and proliferation, whereas suboptimal co-stimulation can often lead to cell death. In recent years, a number of studies have addressed the most effective co-stimulation combinations. It has been demonstrated that CD28 and CD137 costimulation pathways are critical for robust proliferation and cytokine production by T cells [7, 8]. We have tested various combinations of these costimulation pathways to determine the optimal combination for T-cell activation and expansion of CAR-T cells. Here we describe a flow cytometric method to detect successful incorporation and persistence of CAR on the surface of primary T cells after transduction with a retroviral vector carrying CD137 and CD28.
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One of the drawbacks to this approach was that we needed to recompile every time we made even a small change to the code. This slowed everything else we were doing down and because we couldn't always have someone waiting for the application to finish building. 827ec27edc