期刊
CLINICAL AND EXPERIMENTAL IMMUNOLOGY
卷 207, 期 1, 页码 95-103出版社
OXFORD UNIV PRESS
DOI: 10.1093/cei/uxab012
关键词
memory T cell; ex vivo differentiation; naive T cell; CD4; CD8
类别
资金
- Japan Science and Technology Agency (JST) [SHW2019-03]
This study presents a method to culture memory-like T cells from naive human T cells, which involves activation of T cells and subsequent culture under hypoxic conditions. The surviving cells exhibited characteristics of memory T cells and resumed proliferation upon reactivation.
Memory T cells are crucial players in vertebrate adaptive immunity but their development is incompletely understood. Here, we describe a method to produce human memory-like T cells from naive human T cells in culture. Using commercially available human T-cell differentiation kits, both purified naive CD8(+) T cells and purified naive CD4(+) T cells were activated via T-cell receptor signaling and appropriate cytokines for several days in culture. All the T-cell activators were then removed from the medium and the cultures were continued in hypoxic condition (1% O-2 atmosphere) for several more days; during this period, most of the cells died, but some survived in a quiescent state for a month. The survivors had small round cell bodies, expressed differentiation markers characteristic of memory T cells and restarted proliferation when the T-cell activators were added back. We could also induce memory-like T cells from naive human T cells without hypoxia, if we froze the activated T cells or prepared the naive T cells from chilled filter buffy coats. Pre-activated human naive CD4(+) or CD8(+) T cells were cultured in activator-free medium to induce cell death for 5 to 10 days. Although no cell survived in normal oxygen condition (20% O2), 2 to 30% of the activated T cells could survive in hypoxia (1% O2) and became memory-like T cells. We named this simple procedure as 'the Death Assay'.
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