Titlebook: Immune Checkpoint Blockade; Methods and Protocol Yago Pico de Coa?a Book 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Systems-Level Immune Monitoring by Mass Cytometry immune responses in human patients are needed. Mass cytometry allows for detailed profiling of all immune cell populations and their functional responses using a simple blood sample. When combined with appropriate computational analyses, the resolution for distinguishing desired responses from unpr

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Immune Monitoring of Cancer Patients by Multi-color Flow Cytometryo these novel treatments. Multicolor flow cytometry is a powerful tool that enables tumor immunologists to characterize the immune system of patients before and in response to immunotherapy. We present here a protocol for purifying human peripheral blood mononuclear cells and staining them with a se

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High-Throughput, Fast, and Sensitive Immunopeptidomics Sample Processing for Mass Spectrometryand other diseases. So far, methodologies for recovery of HLA class I and II peptides for subsequent mass spectrometry-based analysis have been a major limitation. In this chapter we describe a detailed protocol for a high-throughput, reproducible, and sensitive immunoaffinity-purification of HLA-I

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Expansion of Tumor-Infiltrating Lymphocytes from Melanoma Tumorsom tumors is usually performed in two steps. The details of the procedure differ between different laboratories, but the general concept remains the same. In the first step, small fragments of a tumor are placed in culture medium containing one or more T cell stimulating growth factors. Here the mos

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Establishment of Two Dimensional (2D) and Three-Dimensional (3D) Melanoma Primary Cultures as a Toolottom of plastic flasks. Melanoma cells can be cultured with a considerable degree of success, and, depending on the further use of the cells obtained in the culture, methodologies have to be adjusted to obtain reliable results. Although there are many melanoma continuous cell lines, in vitro 2D and

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Expansion and Determination of Antigen-Reactive T Cells by Flow Cytometryt these cells face major limitations, like the predetermination of the given HLA type. The herein described protocol provides a means of overcoming many of the obstacles associated with the use of multimers and other common approaches in this field. In order to be able to detect rare cells which are