Pro5 - Model MHC -Pentamer

• Uniform bright staining
• Flexible labelling options
• Excellent long term stability
• More than 1800 scientific publications
• Better uniform avidity than MHC tetramers
• Wide range of disease areas and catalog items
• Custom made for the peptides you require
• Application support from the experienced immunology team at ProImmune

Examples of Published Pro5® Pentamer Staining Data:

Collaborative publication by an international team in China and the UK including Oxford University and ProImmune identifying and characterizing SARS-CoV-2 specific T cell responses in convalescent patients:

Peng Y., Dong T., et al. (2020). “Broad and strong memory CD4+ and CD8+ T cells induced by SARS-CoV-2 in UK convalescent COVID-19 patients”

Figure 1. memory phenotype and differentiation status of HLA-B*40:01 restricted NP322-331 (MEVTPSGTWL)-specific T cells. PBMC were isolated and stained with NP/B*40:01 Pentameric complexes and markers of T cell differentiation, and analysed using flow cytometry. A) Expression of differentiation markers of CCR7 and CD45RA on CD8+ Pentamer+ T cells. B): Expression of differentiation markers of CD27 and CD28 on CD8+ Pentamer+ T cells.

In this short clip, Prof. Tao Dong (Professor of Immunology, University of Oxford) describes her latest work in antigen-specific memory T cell responses in SARS-CoV-2 patients, highlighting the recent Nature Immunology publication from her group and the use of Pro5® MHC Pentamers in their work:

If you are interested in Professor Tao Dong’s work and want to hear more, follow this link to our interview with Tao about her career in T cell immunology and recent work with SARS-CoV-2: https://youtu.be/wHIjQn6wwUo

Ferdosi S. R. et. al. (2019). “Multifunctional CRISPR-Cas9 with engineered immunosilenced human T cell epitopes”

Figure 2. Pro5® Pentamer-specific SpCas9 immunodominant epitope-specific CD8+T cell recognition is abolished after anchor residue mutation. a Epitope β-specific CD8+T cell response detected using β-specific Pro5® Pentamer in PBMCs stimulated with peptide β-pulsed antigen presenting cells. b The percentage of CD8+ Pro5® Pentamerβ+T cells was reduced to 0.3% when healthy donor B cell APCs were pulsed with the mutated peptide β2. Table: Positions, sequences, and IEDB HLA binding percentile rank of epitopes α and β before and after mutation of the anchor (2nd and/or 9th) residues. Sb, normalized binding score; Si, normalized immunogenicity score.

Webb J. et. al. (2010). “Profound elevation of CD8+ T cells expressing the intraepithelial lymphocyte marker CD103 (αE/β7 Integrin) in high-grade serous ovarian cancer.”

Figure 3. CD8+ T cells specific for the tumor antigen NY-ESO-1 are predominantly CD103+. Flow cytometry analysis of T cells derived from the malignant ascites of an HLA-A2+ EOC patient (IROC013) who demonstrated reactivity to an HLA-A2-restricted epitope of NY-ESO-1. The analysis shows the frequency of CD8+ lymphocytes staining positive with an A*02:01/SLLMWITQV (NY-ESO-1157–165) Pro5® Pentamer. Pentamer-positive CD8+ T cells were further characterized for CD103 expression (right panel). All events were first gated on total lymphocyte populations by forward and side scatter.

Pro5® Pentamers: consistency by design for the most demanding applications

When you are trying to detect a rare population of antigen specific T cells you will understand the importance of having a research tool that is sensitive, accurate and gives reproducible results. Pro5® MHC Class I Pentamers are designed for these objectives. They bind directly to T cell receptors of a particular specificity, determined by the Major Histocompatibility Complex (MHC) allele and peptide combination. Pro5® Pentamers can be used to detect and separate antigen-specific CD8+ T cell populations as rare as 0.02% of lymphocytes. They are also suitable for detailed epitope characterization and further immune monitoring.

Unlike MHC multimers from other sources, which can be of variable quality and stability, Pro5® Pentamers benefit from a superior design and are made to a professional standard. Each Pentamer is purified four times before being subjected to several stringent quality control steps. Using the Pentamer technology enables you to achieve accurate and reproducible results.

Figure 4. TIGIT expression on CD8+ terminal effector T cells and HIV-specific CD8+ T cells co-stained with various Pro5® MHC Class I Pentamers.

Chew GM et al. (2016) “TIGIT Marks Exhausted T Cells, Correlates with Disease Progression, and Serves as a Target for Immune Restoration in HIV and SIV Infection.” PLoS Pathog 12(1):e1005349. doi:10.1371/journal.ppat.1005349

Praise for Pro5® Pentamers fromDr Cath BollardBaylor College of Medicine, Houston TX, USA

“Pro5® MHC Pentamers have played an important part in our study on adoptive immunotherapy for cancer and viral infections, post-transplant. We use Pentamers to detect and quantify the populations of antigen-specific T cells in CTL lines made for clinical use. We also use them to detect antigen-specific CTL in the peripheral blood of transplant patients, pre and post-CTL-infusion. The Pentamers were chosen for their stability and for the increased intensity and specificity of the positive population compared to MHC tetramers. We have found Pentamers to be consistent and reliable, and ProImmune’s customer service is excellent”.

Pro5® Pentamer Labeling Options

• Pre-labeled with R-PE, APC or biotin Pentamer Fluorotag. Choose this option for ready to go Pentamers for your application.
• Unlabelled, for labeling with R-PE, APC or biotin Pentamer Fluorotag. Choose this option for long-term storage of unlabeled pentamers, projects which require larger batch qualification, changing labels over time, or staining with Pentamer pools.
• Biotin labelled Pentamers allow for unconstrained secondary staining with, e.g. fluochrome-conjugate streptavidin of choice to address any colour or mass cytometry channel.
• Fluorochrome labeled or biotin labeled pentamers are readily usable for bead isolation and subsequent culture of antigen-specific CD8+ T cell.