MHC class I-restricted CD8+ T cell responses are important for controlling virus replication in both human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infection. Since macaque and human MHC class I molecules have been found to bind peptides derived from similar regions of the Gag and Env proteins of HIV and SIV, detailed studies of the rhesus macaque CD8+ T cell response to SIV infection are of considerable benefit to our understanding of the process of HIV infection and successful vaccine development.
In order to enhance SIV research, ProImmune offers Pro5® MHC Class I Pentamers for simian alleles Mamu-A*01 and Mamu-A*02 for use in flow cytometry, to detect and separate antigen-specific CD8+ T cells in the rhesus macaque model. Pro5® Pentamers deliver consistent and reproducible results and are suitable for detailed epitope characterization, as well as analysis on a single cell level when used in combination with intracellular cytokine staining.
ProImmune’s Mamu-A*01 and Mamu-A*02 Pro5® Pentamers were tested by Dr. Marcelo Kuroda at the Tulane National Primate Research Center (USA).
Mamu-A*01-restricted Pro5® Pentamer containing peptide derived from SIV Gag 181-189 (CTPYDINQM), and Mamu-A*02-restricted Pro5® Pentamers containing peptides derived from SIV Nef 159-167 (YTSGPGIRY) and SIV Gag 71-79 (GSENLKSLY), were tested on whole blood from SIVmac251-infected Mamu-A*01+/Mamu-A*02+ rhesus macaques. Cells were incubated with 1 test of R-PE- or APC-labeled Pentamer for 10 minutes at room temperature (22ºC). Following washing, cells were incubated with 1 test anti CD8-PerCP (clone SK1) for 20 minutes at 4ºC. Following a further 2 washes, cells were fixed for analysis by flow cytometry.
The table shows the Pro5® Pentamer specificities that are available as catalog items, with a dispatch time of 5-10 working days. Pro5® Pentamers are available with R-PE, APC or biotin label. Alternatively, an unlabeled Pentamer can be ordered along with a Pro5® Fluorotag (R-PE or APC) or Pro5® Biotag (biotin label). Unlabeled Pentamers are perfect for long term studies as they are guaranteed for 12 months when stored at -80ºC. The corresponding matched peptide can also be ordered in 1 mg or 2 mg quantity.
Alternatively, a Custom Pro5® Pentamer with Mamu-A*01 or Mamu-A*02 can be made to your specification in 4-6 weeks.