Analysis of Tumor-infiltrating Lymphocytes Following CD45 Enrichment

Autor: Rachel Perret, Sophie Sierro, Natalia Botelho, Stephanie Corgnac, Alena Donda, Pedro Romero
Jazyk: angličtina
Rok vydání: 2014
Předmět:
Zdroj: Bio-Protocol, Vol 4, Iss 16 (2014)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.1218
Popis: Measuring antigen-specific T cell responses in the blood and lymphoid organs of vaccinated mice can give us a useful indication of the potency of a vaccine formulation. Unfortunately, systemic or even localized lymphoid T cell responses are not always predictive of the ability of a vaccine to induce tumor protection. Measuring the antigen-specific T cell response within the tumor infiltrating lymphocytes is a more accurate indicator or vaccine efficacy. However, multi-parameter flow cytometric analysis of T cells isolated from tumor tissue can be quite challenging due to the over-whelming number of tumor cells present in relation to the tumor infiltrating lymphocytes (TIL) and to problems associated to the large and adhesive nature of many tumor cell. Here we take advantage of a pre-flow separation of CD45+ leukocytes from the tumor tissue using the MACS magnetic cell sorting system, resulting in a much cleaner cell preparation with which to proceed to flow cytometric staining and analysis.
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