title: Stalk region of beta-chain enhances the coreceptor function of CD8.
abstract: CD8 glycoproteins are expressed as either alphaalpha homodimers or alphabeta heterodimers on the surface of T cells. CD8alphabeta is a more efficient coreceptor than the CD8alphaalpha for peptide Ag recognition by TCR. Each CD8 subunit is composed of four structural domains, namely, Ig-like domain, stalk region, transmembrane region, and cytoplasmic domain. In an attempt to understand why CD8alphabeta is a better coreceptor than CD8alphaalpha, we engineered, expressed, and functionally tested a chimeric CD8alpha protein whose stalk region is replaced with that of CD8beta. We found that the beta stalk region enhances the coreceptor function of chimeric CD8alphaalpha to a level similar to that of CD8alphabeta. Surprisingly, the beta stalk region also restored functional activity to an inactive CD8alpha variant, carrying an Ala mutation at Arg(8) (R8A), to a level similar to that of wild-type CD8alphabeta. Using the R8A variant of CD8alpha, a panel of anti-CD8alpha Abs, and three MHC class I (MHCI) variants differing in key residues known to be involved in CD8alpha interaction, we show that the introduction of the CD8beta stalk leads to a different topology of the CD8alpha-MHCI complex without altering the overall structure of the Ig-like domain of CD8alpha or causing the MHCI to employ different residues to interact with the CD8alpha Ig domain. Our results show that the stalk region of CD8beta is capable of fine-tuning the coreceptor function of CD8 proteins as a coreceptor, possibly due to its distinct protein structure, smaller physical size and the unique glycan adducts associated with this region.
authors.affiliation: Dana-Farber Cancer Institute, Department of Cancer Immunology and AIDS, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Jenny S
authors.initials: JS
authors.lastname: Wong
authors.forename: Xiaosong
authors.initials: X
authors.lastname: Wang
authors.forename: Torsten
authors.initials: T
authors.lastname: Witte
authors.forename: Linghu
authors.initials: L
authors.lastname: Nie
authors.forename: Nicolas
authors.initials: N
authors.lastname: Carvou
authors.forename: Petra
authors.initials: P
authors.lastname: Kern
authors.forename: Hsiu-Ching
authors.initials: HC
authors.lastname: Chang
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017951:Antigen Presentation / Q000235:genetics; D001120:Arginine / Q000235:genetics; D016827:CD8 Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D019281:Dimerization; D006031:Glycosylation; D006183:H-2 Antigens / Q000235:genetics / Q000502:physiology; D007136:Immunoglobulins / Q000378:metabolism / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary / Q000235:genetics; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D011971:Receptors, Immunologic / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000378:metabolism / Q000502:physiology; D014407:Tumor Cells, Cultured
chemical_list: D000276:Adjuvants, Immunologic; D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; C409469:CD8 antigen, alpha chain; C433839:CD8alphabeta antigen; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D007136:Immunoglobulins; D021122:Protein Subunits; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; D001120:Arginine; D000409:Alanine