PepTivator - AdV5 Hexon
Description The PepTivator – AdV5 Hexon is a peptide pool that consists mainly of 15-mer peptides with 11–amino acid overlap. Adenoviruses (AdV) are non-enveloped, ubiquitous, lytic double-stranded DNA viruses. Up to now, 52 different human serotypes (1–52), divided into six species (A-F) are described. The hexon protein is one of the structural proteins in the viral coat and is synthesized during late infection. It is the immunodominant T cell target among capsid proteins and contains multiple epitopes conserved among serotypes. The PepTivator – AdV5 Hexon has been especially developed for efficient in vitro stimulation of AdV5 hexon–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 Applications Quantitative, phenotypical, or functional analysis of AdV5 hexon–specific T cell immunity can provide important information on the natural course of immune responses in healthy or immunocompromised individuals. The in vitro stimulation of AdV5 hexon–specific CD4+ and CD8+ T cells with PepTivator – AdV5 Hexon causes the secretion of effector cytokines and the upregulation of activation markers, such as CD154 or CD137. These cytokines or activation markers then allow the detection and isolation of AdV5 hexon–specific T cells: - Detection and analysis of AdV5 hexon–specific CD4+ and CD8+ effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies. - Isolation of viable AdV5 hexon–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 for research on immunotherapy of AdV infection. - Generation of AdV5 hexon–specific CD4+ and CD8+ effector/memory T cells from naive T cell populations for research on immunotherapy and vaccination. PepTivator – AdV5 Hexon can also be used for pulsing of antigen-presenting cells for research on dendritic cell vaccination.		Figure 1 107 human PBMCs of an AdV+ donor were restimulated for 4 hours with 20 μL/mL of reconstituted PepTivator – AdV5 Hexon. AdV5 hexon–specific cells were stained and magnetically enriched according to their secretion of IFN-γ using the IFN-γ Secretion Assay - Cell Enrichment and Detection Kit. T cells were counterstained for CD4 and CD8 expression. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence. IFN-γ secretion of viable lymphocytes is shown. CD4+ T cells before enrichment after enrichment CD8+ T cells before enrichment after enrichment

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