OMIPs Directory

OMIP-025

Evaluation of human T- and NK-cell responses including memory and follicular helper phenotype by intracellular cytokine staining

Cytometry PART A, Volume 87, Issue 4, 289-292 (2015)
Gemma Moncunill, Carlota Dobano, M. Juliana McElrath, Stephen C. Rosa

Description: This panel was developed to assess antigen-specific T cells using peptide pools to various antigens of interest, although other types of antigens such as recombinant proteins or whole pathogens could be considered using different stimulation times. In addition to multiple functional markers, the panel includes differentiation markers and markers to assess follicular helper T cells and NK cells. It was optimized using cryopreserved peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV) uninfected and HIV infected adults with known cytomegalovirus (CMV) responses and it underwent assay qualification. The panel is being used to evaluate the responses to HIV and malaria vaccine candidates in adults and children from different geographic areas.

Species: Human

Tissue: Peripheral blood

Cell Subset: T cell, T helper, Tfh, T memory, Th1, Th2, Ctl, Treg, Nk

Keyword: Antigen-specific t cell response, Cryopreserved pbmc, Cytomegalovirus (cmv), Hiv, Malaria, Vaccination

Machine: BD LSR2

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
IL-21 Human 3A3-N2 APC Function T cell, Tfh Low
Dead cells Human Live/Dead Fix Aqua viability
CD4 Human SK3 Brilliant Ultraviolet 395 T cell lineage T cell, Tfh, Th1, Th2 High
IFN-gamma Human B27 V450 Function T cell, Th1 High
CD14 Human M5E2 Brilliant Violet 510 monocyte (dump) High
CD40L Human 24-31 Brilliant Violet 605 Function T cell, T memory Med
CD56 Human HCD56 Brilliant Violet 650 NK, NKT-like cells NK Low
IL-4 Human MP4-25D2 PerCP-Cy5.5 Function T cell, Th2 Low
IL-2 Human MQ1-17H12 PE Function T cell, Th1 Med
CD185 Human MU5UBEE PE-eFluor 610 Tfh T cell, Tfh Low
CD45RA Human HI100 APC-H7 memory/differentiation T cell, T memory Med
CD8a Human RPA-T8 Brilliant Violet 711 T cell lineage T cell High
CCR7 Human G043H7 Brilliant Violet 785 memory/differentiation T cell, T memory Low
TNF Human Mab11 FITC Function T cell, Th1 Med
PD-1 Human eBioJ105 (J105) PE-Cy7 Tfh T cell, Tfh Low
CD3 Human UCHT1 Brilliant Violet 570 T cell lineage T cell, Tfh, Th1, Th2 High