Functional activity of staphylococcal enterotoxin A requires interactions with both the alpha and beta chains of HLA-DR.
The staphylococcal enterotoxins, SEA and SEE, bind one zinc atom per molecule of protein. The presence of this metal atom enhances the binding of the toxins to MHC class II molecules, presumably through an interaction with histidine 81 of the beta chain. L cell transfectants expressing HLA-DR1 and HLA-DR7 molecules, with mutations in either the alpha1 or beta1 domains, were tested for their ability to bind SEA and present it to T cells. Cells expressing DR1 molecules with alanine at positions 77, 78, 80, 83, 84 and 85, or serine at position 79 could all bind SEA and present it to either polyclonal or monoclonal T cells. Most point mutations within the alpha-helical portion of the DR7 beta chain had no effect on binding and presentation. However, substitution of histidine 81 with alanine, glutamate, or aspartate, abrogated SEA binding as well as T cell stimulation by the superantigen. This effect was also observed when the non-polymorphic aspartate, at position 76 was changed to alanine. Mutation of the asparagine at position 82 had an intermediate effect. Point mutations of the DR alpha chain had little effect on binding of SEA as determined by a flow cytometric assay. However, mutation of lysine at position 39 of the alpha chain and, to a lesser extent methionine at position 36, markedly decreased the ability of SEA to stimulate toxin-responsive mouse T cell hybridomas. Finally, the monoclonal antibody, L243 binds to the alpha chain of HLA-DR, and was able to block T cell activation by SEA without blocking SEA binding. These data support the model whereby HLA-DR has two binding sites for SEA. A low affinity site, present on the alpha chain, is required for T cell stimulation by the superantigen, but is insufficient to mediate toxin binding. High affinity binding of HLA-DR to SEA occurs solely through residues on the beta chain, including both histidine 81 and aspartate 76.
Dowd
J E
JE
Simmons Arthritis Research Center, UT Southwestern Medical Center, Dallas, TX 75235-8884, USA.
Karr
R W
RW
Karp
D R
DR
D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence / Q000276:immunology; D004768:Enterotoxins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006684:HLA-DR Antigens / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology; D013211:Staphylococcus aureus / Q000276:immunology* / Q000502:physiology*; D018089:Superantigens / Q000737:chemistry / Q000378:metabolism / Q000502:physiology
D004768:Enterotoxins; D006684:HLA-DR Antigens; D018089:Superantigens; C026066:enterotoxin A, Staphylococcal