OMIPs Directory

OMIP-005

Quality and phenotype of antigen-responsive rhesus macaque T cells

Cytometry PART A, Volume 81A, Issue 5, 360-361 (2012)
Kathryn E. Foulds, Mitzi Donaldson, Mario Roederer

Description: The primary eight-color panel was designed to measure IFNγ, IL-2, and TNF production from viable CD4 and CD8 T cells from rhesus macaques in preclinical vaccine studies. An 11-color variant also allows for the assessment of memory subsets based on surface expression of CD28, CD45RA, and CCR7. The panel was optimized not only for use on cryopreserved peripheral blood mononuclear cell (PBMC) samples but also works well on fresh PBMC samples, cryopreserved tissue samples, and fresh tissue samples that have been treated with RBC lysis buffer. The eight-color panel and associated staining procedure were tested in a formal qualification study and shown to be highly reproducible with low inter-aliquot, inter-day, and inter-analyst variability according to the qualification criteria.

Species: Rhesus macaque

Tissue: Peripheral blood

Cell Subset: T cell, T helper, T memory

Keyword: Antigen-specific t cell response, Cryopreserved pbmc, Cryopreserved tissue, Vaccination

Machine: LSR II

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
Dead cells Rhesus macaque Live/Dead Fix Aqua viability
CD45RA Rhesus macaque L48 PE-Cy7 memory T cell, T memory Med
CD28 Rhesus macaque 28.2 PE-Cy5 memory T cell, T memory Med
IFN-gamma Rhesus macaque B27 APC cytokines T cell, T helper High
IL-2 Rhesus macaque MQ1-17H12 PE cytokines T cell, T helper Med
CD8a Rhesus macaque RPA-T8 Pacific Blue T cell T cell, T memory High
CD3 Rhesus macaque SP34-2 APC-Cy7 T cell T cell, T memory, T helper High
CD4 Rhesus macaque S3.5 Qdot 605 T cell T cell, T memory, T helper High
CD69 Rhesus macaque TP1.55.3 ECD background reduction T cell, T memory Med
TNF Rhesus macaque Mab11 FITC cytokines T cell, T memory, T helper High
CCR7 Rhesus macaque 150503 Alexa Fluor 680 memory T cell, T memory Low