Peptide-receptive class I major histocompatibility complex molecules on TAP-deficient and wild-type cells and their roles in the processing of exogenous antigens. 1999

R Song, and A Porgador, and C V Harding
Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA.

These studies addressed the nature and origin of peptide-receptive class I major histocompatibility complex (MHC-I) molecules used to present exogenous antigens. Peptide-receptive Kb molecules in transporter for antigen presentation (TAP)1-/- and TAP1+/+ macrophages were quantitated by exposing cells to exogenous ovalbumin (OVA)(257-264) peptide and then measuring OVA(257-264):Kb complexes with a T hybridoma assay or flow cytometry (using a complex-specific antibody). Relative to TAP1+/+ cells, TAP1-/- cells had decreased levels of pre-existing cell-surface peptide-receptive MHC-I molecules at 37 degrees. With continued exposure of viable cells to peptide, however, TAP1-/- and TAP1+/+ cells formed similar levels of OVA(257-264):Kb complexes, suggesting that nascent labile MHC-I molecules were captured and stabilized by exogenous peptide. Brefeldin A inhibited generation of OVA(257-264):Kb complexes on TAP1-/- (but not TAP1+/+) cells at 37 degrees, confirming the importance of a flux of unstable nascent MHC-I molecules in TAP1-/- cells at 37 degrees. In contrast, at 26 degrees both TAP1-/- and TAP1+/+ cells expressed brefeldin A-resistant, peptide-receptive MHC-I molecules at similar levels. Alternate MHC-I processing of exogenous particulate antigen correlated with ability to present exogenous peptide. Thus, processing was brefeldin A-sensitive with TAP1-/- macrophages at 37 degrees, but brefeldin A-resistant with TAP1+/+ cells at 37 degrees, as well as with TAP1+/+ or TAP1-/- cells at 26 degrees. We conclude that alternate MHC-I antigen processing normally utilizes pre-existing MHC-I molecules, but TAP1-/- cells at 37 degrees mainly use nascent MHC-I molecules, because of a lack of pre-existing, stable, peptide-receptive MHC-I molecules. The results support a vacuolar processing mechanism with binding of peptides to MHC-I molecules in post-Golgi compartments or on the cell surface.

UI MeSH Term Description Entries
D008810 Mice, Inbred C57BL One of the first INBRED MOUSE STRAINS to be sequenced. This strain is commonly used as genetic background for transgenic mouse models. Refractory to many tumors, this strain is also preferred model for studying role of genetic variations in development of diseases. Mice, C57BL,Mouse, C57BL,Mouse, Inbred C57BL,C57BL Mice,C57BL Mice, Inbred,C57BL Mouse,C57BL Mouse, Inbred,Inbred C57BL Mice,Inbred C57BL Mouse
D010047 Ovalbumin An albumin obtained from the white of eggs. It is a member of the serpin superfamily. Serpin B14
D010446 Peptide Fragments Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques. Peptide Fragment,Fragment, Peptide,Fragments, Peptide
D011500 Protein Synthesis Inhibitors Compounds which inhibit the synthesis of proteins. They are usually ANTI-BACTERIAL AGENTS or toxins. Mechanism of the action of inhibition includes the interruption of peptide-chain elongation, the blocking the A site of ribosomes, the misreading of the genetic code or the prevention of the attachment of oligosaccharide side chains to glycoproteins. Protein Synthesis Antagonist,Protein Synthesis Antagonists,Protein Synthesis Inhibitor,Antagonist, Protein Synthesis,Antagonists, Protein Synthesis,Inhibitor, Protein Synthesis,Inhibitors, Protein Synthesis,Synthesis Antagonist, Protein,Synthesis Inhibitor, Protein
D000071181 ATP Binding Cassette Transporter, Subfamily B, Member 2 An ATP-binding cassette, sub-family B protein (ATP BINDING CASSETTE TRANSPORTER, SUBFAMILY B) that functions in the transport of ANTIGENS from the CYTOPLASM to the ENDOPLASMIC RETICULUM for association with HISTOCOMPATIBILITY ANTIGENS CLASS I peptides. It also acts as a molecular scaffold for the final stage of MHC class I PROTEIN FOLDING. ABCB2 Protein,ATP Binding Cassette Transporter, Sub-Family B, Member 2,ATP-Binding Cassette Sub-Family B Member 2,Antigen Peptide Transporter-1,Peptide Transporter PSF1,Peptide Transporter TAP1,RING4 Protein,Really Interesting New Gene 4 Protein,TAP-1 Protein,TAP1 Protein,Transporter 1, ATP-Binding Cassette, Sub-Family B (MDR-TAP),ATP Binding Cassette Sub Family B Member 2,Antigen Peptide Transporter 1,PSF1, Peptide Transporter,TAP 1 Protein,TAP1, Peptide Transporter
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013696 Temperature The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms. Temperatures
D015395 Histocompatibility Antigens Class I Membrane glycoproteins consisting of an alpha subunit and a BETA 2-MICROGLOBULIN beta subunit. In humans, highly polymorphic genes on CHROMOSOME 6 encode the alpha subunits of class I antigens and play an important role in determining the serological specificity of the surface antigen. Class I antigens are found on most nucleated cells and are generally detected by their reactivity with alloantisera. These antigens are recognized during GRAFT REJECTION and restrict cell-mediated lysis of virus-infected cells. Class I Antigen,Class I Antigens,Class I Histocompatibility Antigen,Class I MHC Protein,Class I Major Histocompatibility Antigen,MHC Class I Molecule,MHC-I Peptide,Class I Histocompatibility Antigens,Class I Human Antigens,Class I MHC Proteins,Class I Major Histocompatibility Antigens,Class I Major Histocompatibility Molecules,Human Class I Antigens,MHC Class I Molecules,MHC-I Molecules,MHC-I Peptides,Antigen, Class I,Antigens, Class I,I Antigen, Class,MHC I Molecules,MHC I Peptide,MHC I Peptides,Molecules, MHC-I,Peptide, MHC-I,Peptides, MHC-I
D017737 Macrophages, Peritoneal Mononuclear phagocytes derived from bone marrow precursors but resident in the peritoneum. Peritoneal Macrophages,Macrophage, Peritoneal,Peritoneal Macrophage
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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