CD8 (cluster of differentiation 8) is a cell surface glycoprotein found on most cytotoxic T lymphocytes that functions within the immune system to mediate cell-cell interactions. The functional forms of CD8 consist of either a heterodimer of two isoforms, CD8α and CD8β, or a homodimer of two CD8α molecules. CD8 acts as a coreceptor to facilitate binding between the T-cell Receptor (TCR) and the class I Major Histocompatibility Complex (MHC). Studying CD8 and its importance in isoimmunity can further our understanding of post-transplant recognition.
The CD8a/CD8b (CD8α/CD8β) Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain P2A-linked CD8a (NM_001768.6) and CD8b (NM_004931.5) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1. Schematic of the lenti-vector used to generate the CD8a/CD8b Lentivirus.
Two vials (500 µl x 2) of lentivirus at a titer ≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Formulation
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
UniProt #
CD8a: P01732; CD8b: P10966
Storage/Stability
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Applications
Study CD8-MHC class I interaction.
Generation of a stable cell line expressing CD8a and CD8b with puromycin selection.
Shipping Temperature
-80°C
Notes
To generate a CD8a/CD8b stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve) for antibiotics selection of transduced cells. Visit: https://bpsbioscience.com/cell-line-faq for guidelines on performing a kill curve.
Biosafety The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
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