PepTivator® CMV IE-1 ( #130-093-494 #130-093-493 )

PepTivator® CMV IE-1

Description

The PepTivator^® CMV IE-1 is a peptide pool that consists mainly of 15-mer peptides with 11–amino acid overlap, covering the complete sequence of the cytomegalovirus IE-1 protein.
PepTivator peptide pools have been especially developed for the efficient in vitro stimulation of antigen–specific CD4⁺ and CD8⁺ T cells, as peptides of 15 aa length with 11-aa overlap represent an optimized solution for stimulating both CD4⁺ and CD8⁺ T cells in various applications.¹
Quantitative, phenotypical, or functional analysis of CMV IE-1–specific T cell immunity can provide important information on the natural course of immune responses in healthy or immunocompromised individuals.

Details

Background information
CMV IE-1 (immediate early protein 1), also known as UL123, is a non-structural protein, one of the first CMV antigens expressed in an infected cell, and predominantly induces a CD8⁺ T cell response. IE-1-specific T cells occur in infected individuals at frequencies comparable to those of pp65-specific CD8⁺ T cells.²Both CMV antigens, IE-1 and pp65, are considered dominant T cell targets.

Downstream applications
The in vitro stimulation of IE-1-specific CD4⁺ and CD8⁺ T cells with PepTivator CMV IE-1 causes the secretion of effector cytokines and the upregulation of activation markers, which then allow the detection and isolation of IE-1-specific T cells ³.

Detection and analysis of CMV IE-1-specific CD4⁺ and CD8⁺ effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular staining, or other technologies.
Isolation of viable CMV IE-1-specific CD4⁺ T cells with the CD154 MicroBead Kit, or of CD4⁺ and CD8⁺ T cells using the CD137 MicroBead Kit or MACS Cytokine Secretion Assay - Cell Enrichment and Detection Kits. Subsequently, cells can be expanded for generation of T cell lines.
Generation of CMV IE-1-specific CD4⁺ and CD8⁺ effector/memory T cells from naive T cell populations.
Pulsing of antigen-presenting cells for research on dendritic cell vaccination.

Figure 1

From a CMV⁺ donor 10⁶ human PBMCs were restimulated for 6 hours with 20 μL/mL of reconstituted PepTivator CMV IE-1 or incubated without antigen. After 2 hours 1 μg/mL brefeldin A was added. Cells were fixed, permeabilized, and intracellularly stained with Anti-IFN-γ-PE. T cells were counterstained for CD8 expression.

Stimulated sample

Unstimulated control