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The B cell line 721.174 has lost the ability to present intracellular antigens to major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocytes (CTL). This phenotype results from a homozygous deletion in the MHC that includes the peptide transporter genes TAP1 and TAP2, and the proteasome subunits LMP2 and LMP7. Recent work has shown that such cells transfected with TAP genes load their class I molecules with endogenous peptides, and present several viral epitopes to class I-restricted CTL. These data implied that the LMP2 and LMP7 genes were not required for the presentation of most epitopes through class I molecules. By contrast, while confirming the previous reports, we have identified several epitopes that appear to require genes in the MHC in addition to the TAP for their presentation. Further analysis localizes the defect to proteolysis in the cytosol. In one case, presentation could be partially restored by re-expression of full-length LMP7. Control experiments with LMP7, from which the putative pro-region had been removed, failed to restore presentation, and this lack of effect correlated with failure of the shortened LMP7 to incorporate into the proteasome. These results suggest a role for LMP7 in the generation of a viral epitope, but leave open the possibility that additional genes within the .174 deletion are required for full restoration of antigen presentation.

Original publication

DOI

10.1002/eji.1830250238

Type

Journal article

Journal

Eur J Immunol

Publication Date

02/1995

Volume

25

Pages

554 - 562

Keywords

ATP-Binding Cassette Sub-Family B Member 2, ATP-Binding Cassette Transporters, ATP-Binding Cassette, Sub-Family B, Member 3, Amino Acid Sequence, Animals, Antigen Presentation, Base Sequence, Cell Line, Cysteine Endopeptidases, H-2 Antigens, Histocompatibility Antigen H-2D, Humans, Major Histocompatibility Complex, Molecular Sequence Data, Multienzyme Complexes, Proteasome Endopeptidase Complex, Proteins, Rats, Transfection, Vaccinia virus