Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 Nov 4;286(44):38054-38059.
doi: 10.1074/jbc.M111.281808. Epub 2011 Sep 13.

Allele-dependent processing pathways generate the endogenous human leukocyte antigen (HLA) class I peptide repertoire in transporters associated with antigen processing (TAP)-deficient cells

Elena Lorente  1 , Ruth García  1 , Daniel López  2
Affiliations

Allele-dependent processing pathways generate the endogenous human leukocyte antigen (HLA) class I peptide repertoire in transporters associated with antigen processing (TAP)-deficient cells

Elena Lorente et al. J Biol Chem. .

Abstract

The transporters associated with antigen processing (TAP) allow the supply of peptides derived from the cytosol to translocate to the endoplasmic reticulum, where they complex with nascent human leukocyte antigen (HLA) class I molecules. However, infected and tumor cells with TAP molecules blocked or individuals with nonfunctional TAP complexes are able to present HLA class I ligands generated by TAP-independent processing pathways. These peptides are detected by the CD8(+) lymphocyte cellular response. Here, the generation of the overall peptide repertoire associated with four different HLA class I molecules in TAP-deficient cells was studied. Using different protease inhibitors, four different proteolytic specificities were identified. These data demonstrate the different allele-dependent complex processing pathways involved in the generation of the HLA class I peptide repertoire in TAP-deficient cells.

PubMed Disclaimer

Figures

FIGURE 1.
FIGURE 1.
Surface reexpression of HLA class I molecules after acid stripping in the presence of BFA. T2 and T2-B27 cells were untreated (black bars) or incubated with 5 μg/ml BFA for 6 h (hatched bars) after acid washing. Stability at the cell surface of HLA-A2, -B27, -B51, and -Cw1 class I molecules of the TAP-deficient cells was measured by flow cytometry using monoclonal Ab ME1 (anti-HLA-B27) in T2-B27 cells; polyclonal H00003106-B01P (specific for HLA-B class I molecules) in T2 cells; and monoclonal Abs PA2.1 (anti-HLA-A2) and polyclonal SC-19438 (specific for HLA-C class I molecules) in T2 and T2-B27 cells simultaneously. The data are expressed as MFI ± S.D. (error bars) (upper) or percentage of inhibition ± S.D. (lower) of HLA surface reexpression in presence of BFA and are the means of three or four different experiments.
FIGURE 2.
FIGURE 2.
Effect of several proteasome inhibitors on surface reexpression of HLA class I molecules after acid washing. T2 and T2-B27 cells as in Fig. 1 were incubated with lactacystin (open bars) or epoxomicin (filled bars) at the indicated concentrations (Table 1) as in Fig. 1. The data are expressed as percentage of inhibition ± S.D. (error bars) as in Fig. 1 and are the means of four or five different experiments.
FIGURE 3.
FIGURE 3.
Surface reexpression of HLA class I molecules after acid stripping in the presence of several protease classes inhibitors. Upper, T2 and T2-B27 cells as in Fig. 1 were incubated with the indicated inhibitors at the concentrations summarized in Table 1. HLA-A2 (filled bars), -B27 (open bars), -B51 (right hatched bars), and -Cw1 (left hatched bars) surface reexpression was measured. The data are expressed as percentage of inhibition ± S.D. (error bars) as in Fig. 1 and are the means of three to six different experiments. Lower, a representative experiment with T2 cells stained with anti-HLA-A2 Ab is depicted. The code used is as follows: shaded histogram, second Ab alone (negative control); thin line, no inhibitor; and thick line, 30 μm leucinethiol.
FIGURE 4.
FIGURE 4.
Outcome of different protease-specific inhibitors or a lysosomotropic agent on surface reexpression of HLA class I molecules after acid washing. Upper, T2 and T2-B27 cells as in Fig. 1 were incubated with the indicated drugs at the concentrations summarized in Table 1. HLA-A2 (filled bars), -B27 (open bars), -B51 (right hatched bars), and -Cw1 (left hatched bars) surface reexpression was calculated. The data are expressed as percentage of inhibition ± S.D. (error bars) as in Fig. 1 and are the means of three to six different experiments. Lower, a representative experiment with T2 cells stained with anti-HLA-A2 Ab is depicted. The code used as in follows: shaded histogram, second Ab alone; thin line, no inhibitor; and thick line, 100 μm z-LL2.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Seifert U., Marañón C., Shmueli A., Desoutter J. F., Wesoloski L., Janek K., Henklein P., Diescher S., Andrieu M., de la Salle H., Weinschenk T., Schild H., Laderach D., Galy A., Haas G., Kloetzel P. M., Reiss Y., Hosmalin A. (2003) Nat. Immunol. 4, 375–379 - PubMed
    1. Guil S., Rodríguez-Castro M., Aguilar F., Villasevil E. M., Antón L. C., Del Val M. (2006) J. Biol. Chem. 281, 39925–39934 - PubMed
    1. York I. A., Bhutani N., Zendzian S., Goldberg A. L., Rock K. L. (2006) J. Immunol. 177, 1434–1443 - PubMed
    1. Stoltze L., Schirle M., Schwarz G., Schröter C., Thompson M. W., Hersh L. B., Kalbacher H., Stevanovic S., Rammensee H. G., Schild H. (2000) Nat. Immunol. 1, 413–418 - PubMed
    1. Parmentier N., Stroobant V., Colau D., de Diesbach P., Morel S., Chapiro J., van Endert P., Van den Eynde B. J. (2010) Nat. Immunol. 11, 449–454 - PubMed

Publication types

MeSH terms