The binding affinity of the pMHCI–CD8 interaction, measured by surface plasmon resonance, is largely conserved across the majority of MHCI allotypes studied to date (Tables 1a–c). Notably, the average human pMHCI–CD8αα interaction exhibits very low solution binding affinities (average KD = 145 μm) in a relatively tight range (KD = 100–220 μm) (Table 1a)

and is characterized by extremely rapid kinetics (Koff > 18 s−1).[36, selleck inhibitor 37] There are, however, some exceptions to this overall uniformity. For example, HLA-A*6801 and HLA-B*4801 contain A245V and A245T mutations, respectively, in their α3 domains that substantially reduce CD8 binding (KD ∼ 1000 μm) (Table 1a).[38] The biology that underlies these anomalies remains poorly defined, although the fact that CD8 can still bind, albeit with very low binding affinity, is likely to be important to impose MHCI restriction Aloxistatin mw upon T cells restricted by these alleles.[34] Furthermore, the extremely weak binding affinity of CD8 to HLA-A*6801 still allows most of the benefits, in terms of antigen recognition, that are seen with the wild-type interaction.[38] In the murine system, affinity measurements have been reported for CD8αα and CD8αβ binding to a range of different MHCI alleles (Table 1b,c).

The average binding affinity for CD8αα (KD = 69 μm) is similar to that of CD8αβ (KD = 49 μm) despite the small structural differences reported for pMHCI–CD8αα and pMHCI–CD8αβ,[29] but the range of affinity measurements is somewhat larger than in the human system (CD8αα KD = 6·7–210 μm and CD8αβ KD = 14·1–135 μm). Hence, unlike in the human system, there seems to be some substantial differences in binding affinity between alleles. However, this observation should be considered with caution as there are inconsistencies for some measurements. For example, the interaction between CD8αβ and H2-Db has been measured by one group as KD = 14·1 μm [39] and by another group as KD > 1000 μm.[40] The H2-Db molecules used in these separate experiments were complexed to different peptides, raising the possibility that peptide-induced modulation

of CD8 binding could be at play. However, there has been no evidence in Astemizole any other MHCI system to suggest that the bound peptide can affect CD8 binding, hence it is possible that differences in protein synthesis and experimental design may have had some impact on these disparate findings. Nonetheless, it is clear that CD8 operates at a very weak binding affinity compared with the TCR in both the human and murine systems. Although pMHCI–CD8 binding affinity measurements have shown that the interaction is weak, there is potential for CD8 to bind to pMHCI simultaneously with the TCR. This begs the question of whether the TCR, or CD8, binds more strongly to pMHCI during TCR–pMHCI–CD8 tripartite complex formation compared with the dipartite interactions.