OMIP-062
A 14‐Color, 16‐Antibody Panel for Immunophenotyping Human Innate Lymphoid, Myeloid and T Cells in Small Volumes of Whole Blood and Pediatric Airway Samples
Cytometry PART A, Volume 95, Issue 12, 1231-1235 (2019)
Dawid Swieboda, Yanping Guo, Sophie Sagawe, Ryan S. Thwaites, Simon Nadel, Peter J.M. Openshaw, Fiona J. Culley
Description: This 14‐color, 16‐antibody OMIP was designed for enumeration of leukocyte responses in pediatric samples, where sample volumes and cell numbers can be very low. Leukocytes identified by this panel include all major members of the innate lymphoid cell (ILC) family (ILC1s, ILC2s, and ILC3s), natural killer cells (NK cells), granulocytes (neutrophils and eosinophils), T‐cells (CD4+ and CD8+), mucosal‐associated invariant T cells (MAIT cells) and NKT‐like cells. The protocol was optimized using small volumes of peripheral blood and validated in airway samples obtained from children (< 2 years of age) admitted to a pediatric intensive care unit (PICU). Given this backdrop, this OMIP is widely applicable to clinical research using low volume or paucicellular samples, such as studies of innate and adaptive immune responses in infants and children, with potential clinical application in diagnostics and monitoring of patients by pediatricians.
Species: Human
Tissue: Cord blood, Nasal aspirate, Peripheral blood, Tracheal aspirate
Cell Subset: T cell, Mait, Ilc, Nk, Nkt, Granulocyte, Eosinophil, Neutrophil
Keyword: Pediatric
Machine: BD LSR Fortessa
Panel details
Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
---|---|---|---|---|---|---|
CD14 | Human | 63D3 | Brilliant Violet 510 | Lineage | High | |
CD19 | Human | HIB19 | Brilliant Violet 510 | Lineage | Low | |
CD123 | Human | 6H6 | Brilliant Violet 510 | Lineage | Med | |
CD4 | Human | RPA-T4 | Brilliant Violet 605 | CD4+ T cells | T cell | High |
TCR V alpha 7.2 | Human | 3C10 | Brilliant Violet 785 | MAIT cells | MAIT | Med |
CD45 | Human | HI30 | FITC | Leukocyte | ILC, T cell, Neutrophil, NK | High |
CD117 | Human | A3C6E2 | PerCP-Cy5.5 | ILC3 | ILC | Med |
CD3 | Human | OKT3 | PE | T cells | T cell | High |
CD161 | Human | HP-3G10 | PE-Dazzle 594 | MAIT cells | MAIT, ILC | Low |
CD56 | Human | 5.1H11 | PE-Cy7 | NK/NKT-like cells | NK | Low |
CD294 | Human | BM16 | Alexa Fluor 647 | ILC2/Th2/Tc2 subsets | ILC | Med |
CD66b | Human | G10F5 | Alexa Fluor 700 | Eosinophils | Neutrophil | High |
Dead cells | Human | Live/Dead Fix Near IR (775) | Viability | |||
Fc epsilon R1 alpha | Human | AER-37 (CRA-1) | Brilliant Violet 510 | Lineage | High | |
CD8a | Human | SK1 | Brilliant Violet 711 | CD8+ T cells | T cell | High |
CD16 | Human | 3G8 | Brilliant Violet 650 | NK cells/neutrophils | NK, Neutrophil | Med |
CD127 | Human | A019D5 | Brilliant Violet 421 | ILCs | ILC | Med |