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PepTivator
®
BKV VP1
PepTivator
®
BKV VP1
Stimulate T cells reactive to the VP1 capsid protein of the BK virus (BKV) using PepTivator Peptide Pools. PepTivators are pools of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap. PepTivator
®
BKV VP1 is a pool of lyophilized peptides, covering the sequence of the BKV VP1 capsid protein (UniProt ID: P14996).
The PepTivator is available in two different quality grades. The research-grade product has an average purity of 70%, whereas peptides of the premium-grade product are individually purified via HPLC having each a purity of >80%.
In vitro
stimulation of antigen-specific T cells with PepTivator Peptide Pools causes the secretion of
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
+
and CD8
+
T cells.
Specifications for
PepTivator
®
BKV VP1
Overview
Stimulate T cells reactive to the VP1 capsid protein of the BK virus (BKV) using PepTivator Peptide Pools. PepTivators are pools of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap. PepTivator
®
BKV VP1 is a pool of lyophilized peptides, covering the sequence of the BKV VP1 capsid protein (UniProt ID: P14996).
The PepTivator is available in two different quality grades. The research-grade product has an average purity of 70%, whereas peptides of the premium-grade product are individually purified via HPLC having each a purity of >80%.
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.
Detailed product information
Background information
BK virus (BKV) is a ubiquitous polyomavirus closely related to the JC polyomavirus (JCV). After primary infection of humans, BKV persists in a latent state especially in the kidney and the urinary tract. In healthy individuals the infection is asymptomatic, but in immunocompromised patients virus reactivation may occur, which then can cause, e.g., haemorrhagic cystitis after allogenic stem cell transplantation or loss of the allograft after renal transplantation. VP1 is the major capsid protein of BKV, expressed late in the lytic cycle and a target of T cell immunity.
Downstream applications
PepTivator BKV VP1 has been specifically developed for efficient
in vitro
stimulation of BKV VP1–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 VP1–specific effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
Isolation of viable BKV VP1–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 VP1–specific effector/memory T cells from naive T cell populations.
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Detection and isolation of viable BKV–specific T cells by intracellular staining using MACS® Rapid Cytokine Inspector (CD4/CD8 T Cell) Kit (# 130-097-343)
From a BKV
+
donor, 10
6
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). PBMCs stimulated with PepTivator CEF MHC Class I Plus (# 130-098-426) were used as positive control.
The cells were stained with the CD4/CD8 T Cell Detection Cocktail and Rapid Cytokine Inspector Anti-IFN-γ-PE as described and were analyzed by flow cytometry using the MACSQuant
®
Analyzer. Cell debris, monocytes, and B cells were excluded from the analysis. IFN-γ expression was analyzed for CD4
Stimulate T cells reactive to the VP1 capsid protein of the BK virus (BKV) using PepTivator Peptide Pools. PepTivators are pools of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap. PepTivator
®
BKV VP1 is a pool of lyophilized peptides, covering the sequence of the BKV VP1 capsid protein (UniProt ID: P14996).
The PepTivator is available in two different quality grades. The research-grade product has an average purity of 70%, whereas peptides of the premium-grade product are individually purified via HPLC having each a purity of >80%.
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.
Detailed product information
Background information
BK virus (BKV) is a ubiquitous polyomavirus closely related to the JC polyomavirus (JCV). After primary infection of humans, BKV persists in a latent state especially in the kidney and the urinary tract. In healthy individuals the infection is asymptomatic, but in immunocompromised patients virus reactivation may occur, which then can cause, e.g., haemorrhagic cystitis after allogenic stem cell transplantation or loss of the allograft after renal transplantation. VP1 is the major capsid protein of BKV, expressed late in the lytic cycle and a target of T cell immunity.
Downstream applications
PepTivator BKV VP1 has been specifically developed for efficient
in vitro
stimulation of BKV VP1–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 VP1–specific effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
Isolation of viable BKV VP1–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 VP1–specific effector/memory T cells from naive T cell populations.
