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OMIP-009

Characterization of antigen-specific human T-cells

Laurie Lamoreaux, Richard A. Koup, Mario Roederer Cytometry PART A, Volume 81A, Issue 5, 363-363 (2012)

PURPOSE: The panels described in this article are designed to characterize the immunological response of human T-cells to vaccination by measuring the frequency, phenotype, and function of CD4 and CD8 T-cells. Subsequent qualification of the panel allows for comparison of intra- and inter-laboratory outcomes between different vaccine trials such that those vaccine formulations that reveal a possible correlate of protection against infection may be moved forward in the regulatory process. Although the panel was developed for batch analysis of cryopreserved PBMC samples, the assay may also be performed with fresh cells (Table 1).

CELL TYPE: Cryopreserved or Fresh Peripheral Blood Mononuclear Cells

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human N/A Live/Dead Fix Violet viability
CD3 Human SP34-2 APC-Cy7 lineage
CD45RA Human L48 PE-Cy7 memory/differentiation
CD28 Human PE-Cy7 memory/differentiation
ccr7 Human 150503 Alexa Fluor 680 memory/differentiation
CD4 Human T4 ECD lineage
CD8 Human RPA-T8 Pacific Blue lineage
IFN-gamma Human B27 APC function
IL-2 Human MQ1-17H12 PE function
TNF alpha Human Mab11 FITC function