PepTivator® EBV LMP2A ( #130-093-615 #130-093-616 )

PepTivator® EBV LMP2A

Description

PepTivator^® EBV LMP2A is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the sequence of the Epstein-Barr virus LMP2A protein.
PepTivator peptide pools have been developed for the efficient stimulation of CD4⁺ and CD8⁺ T cells, as peptides of 15 aa length and 11 aa overlap represent the optimal solution for stimulating both, CD4⁺ and CD8⁺ T cells in various applications.
Quantitative, phenotypical, or functional analysis of EBV LMP2A–specific T cell immunity can provide important information on the natural course of immune responses in healthy or immunocompromised individuals.

Details

Background information
Epstein-Barr virus (EBV) is a human γ-herpesvirus with B cell growth–transforming ability and lymphomagenic potential. More than 90% of human adults are infected with EBV. In healthy individuals, EBV typically establishes a persistent latent infection, in which the virus can be detected in resting, non-proliferating peripheral B lymphocytes. Expression of the viral protein latent membrane protein 2A (LMP2A) maintains the latency phase.
T cells are essential for the control of the outgrowth of EBV-infected B cells and LMP2A is one of the immunogenic targets. In EBV-associated malignancies, such as nasopharyngeal carcinoma and Hodgkin’s lymphoma, LMP2A is one of a restricted number of expressed viral proteins.¹

Downstream applications
The in vitro stimulation of LMP2A–specific CD4⁺ and CD8⁺ T cells with PepTivator EBV LMP2A causes the secretion of effector cytokines and the up-regulation of activation markers, which then allow the detection and isolation of LMP2A–specific T cells.

Detection and analysis of LMP2A–specific CD4⁺ and CD8⁺ effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
Isolation of viable LMP2A–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 LMP2A–specific CD4⁺ and CD8⁺ effector/memory T cells from naive T cell populations for research on immunotherapy and vaccination.
Pulsing of antigen-presenting cells for research on dendritic cell vaccination.

PBMCs from an EBV⁺ donor were restimulated for 6 hours with 20 μL/mL of PepTivator EBV LMP2A. After 2 hours, Brefeldin A was added. Cells were fixed, permeabilized, and LMP2A–specific cells were intracellularly stained with Anti-IFN-γ-PE. T cells were counterstained for CD4 and CD8 expression. IFN-γ production of lymphocytes is shown.

CD4⁺ T cells

Stimulated sample

Non-stimulated control

CD8⁺ T cells

Stimulated sample

Non-stimulated control