OMIPs Directory

OMIP-080

29-Color flow cytometry panel for comprehensive evaluation of NK and T cells reconstitution after hematopoietic stem cells transplantation

Cytometry PART A, Volume 101, Issue 1, 21-26 (2022)
Sarka Vanikova,Abhishek Koladiya,Jan Musil

Description: This 29-color panel was developed and optimized for the monitoring of NK cell and T cell reconstitution in peripheral blood of patients after HSCT. We considered major post-HSCT complications during the design, such as relapses, viral infections, and GvHD and identification of lymphocyte populations relevant to their resolution. The panel includes markers for all major NK cell and T cell subsets and analysis of their development and qualitative properties. In the NK cell compartment, we focus mainly on CD57 + NKG2C+ cells and the expression of activating (NKG2D, DNAM-1) and inhibitory receptors (NKG2A, TIGIT). Another priority is the characterization of T cell reconstitution; therefore, we included detection of CD4+ RTEs based on CD45RA, CD62L, CD95, and CD31 as a marker of thymus function. Besides that, we also analyze the emergence and properties of major T cell populations with a particular interest in CD8, Th1, ThCTL and Treg subsets. Overall, the panel allows for comprehensive analysis of the reconstituting immune system and identification of potential markers of immune cell dysfunction.

Species: Human

Cell Subset: T cell, Ctl, T helper, T memory, Tcrgd, Th1, Treg, Nk, Recent thymic emigrants (rte)

Keyword: Activation, Allogeneic stem cell transplantation, Graft versus host disease (gvhd), Immune checkpoint

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD8a Human RPA-T8 Brilliant Ultraviolet 805 CD8 T cell and NKT-like cell lineage marker T cell, T memory High
Live/Dead Fix Blue All Species Live/Dead Fix Blue Viability
CD69 Human FN50 Brilliant Ultraviolet 496 NK and T cell activation marker T cell, NK, Thelper, Treg, TCRgd Med
CCR6 Human 11A9 Brilliant Ultraviolet 563 Chemokine receptor; Th subset identification T helper, T cell Med
CD27 Human M-T271 Brilliant Ultraviolet 615 T cell differentiation T cell, T memory, Thelper, Treg, TCRgd Med
PD-1 Human EH12.1 Brilliant Ultraviolet 661 T cell inhibitory receptor T cell Low
CD45RA Human HI100 Pacific Blue T cell differentiation T cell, T memory, TCRgd Med
CD366 (TIM-3) Human 7D3 Brilliant Violet 480 T cell inhibitory receptor T cell Low
CD4 Human RPA-T4 Brilliant Violet 570 CD4 T cell lineage marker T cell, T memory, Thelper, Treg High
TCR gamma/delta Human 11F2 Brilliant Violet 711 γδ T cells TCRgd Low
CD226 Human DX11 Brilliant Violet 750 T cell and NK cell activating receptor T cell, NK Med
CD31 Human WM59 Brilliant Violet 786 Adhesion molecule; identification of RTE High
CCR10 Human 1B5 Brilliant Blue 515 Chemokine receptor; Th subset identification Thelper, T cell Low
CD194 Human [1G1] Brilliant Blue 700 Chemokine receptor; Th subset identification T cell, T helper Med
TIGIT Human 741182 Brilliant Blue 755-P T cell and NK cell inhibitory receptor T cell, NK Low
CD314 (NKG2D) Human 1D11 Brilliant Blue 790-P NK cell activating receptor NK Low
CD56 Human NCAM16.2 PE-CF594 NK cell and NKT-like cell lineage marker NK Low
CD183 (CXCR3) Human 1C6/CXCR3 PE-Cy5 Chemokine receptor; Th subset identification T cell, Thelper Med
CD3 Human UCHT1 Alexa Fluor 647 T cell and NKT-like cell lineage marker T cell, Treg, Thelper, T memory High
CD16 Human 3G8 Alexa Fluor 700 NK cell differentiation NK Med
CD159a (NKG2A) Human 131411 Brilliant Violet 421 NK and NKT-like cell inhibitory receptor NK Low
CD95 Human Dx2 Brilliant Violet 650 T cell activation and differentiation T cell, T memory Low
CD159c Human 134591 PE NK cell activating receptor NK Med
CD39 Human A1 PE-Cy7 Treg activation marker T cell, Treg Med
Perforin Human B-D48 APC-Fire 750 Cytolytic function T cell, NK High
CD25 Human 2A3 Brilliant Ultraviolet 737 T cell activation; Treg identification T cell, Treg Low
CD62L Human Dreg-56 Brilliant Ultraviolet 395 T cell differentiation T memory, NK, Treg, Thelper, T cell, TCRgd Med
CD57 Human QA17A04 Brilliant Violet 605 T cell and NK cell differentiation NK, T cell, TCRgd High
FOXP3 Human PCH101 PE-Cy5.5 Master transcription factor for Tregs T cell, Treg Low