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Immunologic checkpoint expression on murine effector and memory T-cell subsets

Satoshi Nemoto, Adam W. Mailloux, Jodi Kroeger, James J. Mulé Cytometry PART A, Volume 89, Issue 5, 427–429 (2016)

PURPOSE: This panel was designed to assess the expression levels of cell surface inhibitory receptors known as “immune checkpoints” within the context of multiple naïve, activated, memory, and effector phenotypes among T-cells for subsequent adoptive transfer using the CD45.1/CD45.2 congenic system in C57BL/6 mice. It can be easily adapted to other congenic systems, or may be used without any congenic marker. While many panels have been published that analyze T-cell activation, memory phenotypes, or effector differentiation states, few, if any, are comprehensive enough to assess these compartments simultaneously while measuring inhibitory immune checkpoint receptor expression. The ability to do so within a congenic system creates a powerful tool for investigating the evolution of T-cell based immune responses in a broad range of contexts. Here, the panel is used to analyze the T-cell compartment in normal spleen, or T-cells infiltrating subcutaneous murine colon adenocarcinoma, MC38. However, any murine source of T-cells would serve as an appropriate sample source for this panel.

CELL TYPE: Any source containing murine T-cells


Marker Target Clone Fluorochrome Purpose
CD3 Mouse 145-2C11 BUV395 T-cell
CD4 Mouse Gk1.5 BUV805 TH
CD8 Mouse 53-6.7 Alexa Fluor 700 TC
CD69 Mouse H1.2F3 PE-CF594 Activation
CD44 Mouse IM7 Alexa Fluor 488 Memory
CD45RA Mouse 14.8 Brilliant Violet 786 Memory
CD27 Mouse LG.3A10 Brilliant Violet 510 Memory
CD62L Mouse Mel-14 PE-Cy7 Memory
KLRG1 Mouse 2F1/KLRG1 PerCP-Cy5.5 Differentiation
CD127 Mouse SB/199 BUV737 Differentiation
PD-1 Mouse J43 Brilliant Violet 605 Checkpoint
CTLA-4 Mouse UC10-4B9 PE Checkpoint
TIM-3 Mouse B8.2C12 APC Checkpoint
Lag-3 Mouse C9B7W Brilliant Violet 711 Checkpoint
CD45.2 Mouse 104 APC-Cy7 Congenic
Dead cells Mouse DAPI Viability