PepTivator® BKV LT is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the complete sequence of the human BK virus (BKV) large T (LT) antigen (UniProt ID: P14999).
The in vitro stimulation of antigen-specific T cells with PepTivator Peptide Pools causes the secretion of effector cytokines and the up-regulation of activation markers, which then allow the detection and isolation of antigen-specific T cells.
BKV is a ubiquitous polyomavirus. After primary infection BKV persists in a latent state. Virus reactivation might occur in immunodeficiency or during immunosuppression. The LT antigen is expressed early in the lytic cycle and is involved in virus replication.
PepTivator BKV LT - research grade has been specifically developed for efficient in vitro stimulation of BKV LT–specific T cells. Peptides of 15 amino acids in length and 11 amino acids overlap represent an optimized solution for stimulating both CD4+ and CD8+ T cells in various applications, including:
- Detection and analysis of BKV LT–specific effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
- Isolation of viable BKV LT–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 BKV LT–specific effector/memory T cells from naive T cell populations.
- Pulsing of antigen-presenting cells.
Detection and isolation of viable BKV–specific T cells using MACS® IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE)
From a BKV+ donor, 107 human peripheral blood mononuclear cells (PBMCs) were restimulated for 4 hours either with 20 μL/mL of reconstituted PepTivator® BKV LT1 (A), 20 µL/mL BKV VP1 (B), or a mixture of both peptide pools (C). BKV-specific T cells were stained and magnetically enriched according to their secretion of IFN-γ using the IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE) (# 130-054-201). T cells were counterstained for CD4 and CD8 expression. Cell debris and dead cells are excluded from the analysis based on scatter signals and propidium iodide fluorescence. IFN-γ secretion of viable lymphocytes is shown.