OMIP‐050: A 28‐color/30‐parameter Fluorescence Flow Cytometry Panel to Enumerate and Characterize Cells Expressing a Wide Array of Immune Checkpoint Molecules

By Leonard Nettey, Amber J. Giles, Pratip K. Chattopadhyay. 2018

44 citations recorded by the source corpus.

This 28-color/30-parameter panel catalogs, enumerates and characterizes cells expressing molecules that regulate T-cell responses (“checkpoint” molecules). The primary purpose is to measure combinatorial expression of PD-1, CTLA-4, TIM-3, CD244/2B4, TIGIT, BTLA, CD137/4-1BB, GITR, OX40, CD27, and CD278/ICOS, many of which are targets of the immunotherapy drugs in preclinical or clinical development. This panel is optimized for cryopreserved healthy human peripheral blood mononuclear cells (PBMCs) and cryopreserved mononuclear cells isolated from tumor specimens (tissue MNCs). CD45 is used to distinguish leukocytes from stromal cells and debris often present in tissue digests. CD3, CD4, and CD8 identify T-cells, whose differentiation status is then assessed with CD45RO, CD95, CCR7, CD27, CD57, and CD28. CD25, HLA-DR, and CD69 mark activated T-cells. CD69 is also used to identify tissue-resident lymphocytes in combination with CD103. CXCR3 and CXCR6 mediate lymphocyte trafficking toward chemokines often found in the tumor microenvironment. Regulatory T-cells are defined here by antibodies against CD25 and CD127. In sum, this panel comprehensively characterizes cells expressing checkpoint markers.

Subfields: Immunology, Oncology

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In Immunology

1. Josef Spidlen
2. Richard L. Halpert
3. Chris Ciccolella
4. R. Anno
5. J. Snyder-Cappione
6. Anna C. Belkina

In Oncology

1. Josef Spidlen
2. Richard L. Halpert
3. Chris Ciccolella
4. R. Anno
5. J. Snyder-Cappione
6. Anna C. Belkina

Citations in the corpus

• Automated Optimal Parameters for T-Distributed Stochastic Neighbor Embedding Improve Visualization and Allow Analysis of Large Datasets (2018, background): “With recent advances in instrumentation and reagent availability, flow cytometry datasets with >20 parameters are quickly becoming prevalent and even standard in the field 29-31 , yet the proper data assessment tools are lacking for general use.”