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. 2020 Oct 9;8(2):e000706. doi: 10.1136/jitc-2020-000706

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© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

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Consistent liberation of viable CD8+ effector memory T cells from clear cell renal cell carcinoma (ccRCC). (A) Creation of a fresh tumor digest (FTD) from ccRCC. Green arrows indicate viable presumptive lymphocytes, and red arrows indicate viable presumptive tumor cells after trypan blue exclusion. (B) Representative CD4+ and CD8+ staining of pre-expansion CD3+ RCC tumor-infiltrating lymphocytes (TILs). (C) Frequencies of pre-expansion CD8+, CD4+, double-negative (DN), and double-positive (DP) T cells from 10 ccRCC samples. (D) Representative effector (eff), effector memory (em), central memory (cm) and stem-cell like memory/naive (scm/nv) staining of pre-expansion CD3+/CD8+ RCC TILs. (E) Frequencies of pre-expansion CD3+/CD8+ with an eff, em, cm, and scm/nv phenotype from 10 ccRCC samples. (F) Outline of the three TIL expansion methods used. ACD, adherent cell depletion; ACK, ammonium chloride–potassium; IL-2, aldesleukin; PreREP, Pre-Rapid Expansion Protocol.