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. 1998 Apr 14;95(8):4516-21.
doi: 10.1073/pnas.95.8.4516.

Cathepsins B and D are dispensable for major histocompatibility complex class II-mediated antigen presentation

J Deussing  1 , W Roth, P Saftig, C Peters, H L Ploegh, J A Villadangos
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Cathepsins B and D are dispensable for major histocompatibility complex class II-mediated antigen presentation

J Deussing et al. Proc Natl Acad Sci U S A. .

Abstract

Antigen presentation by major histocompatibility complex (MHC) class II molecules requires the participation of different proteases in the endocytic route to degrade endocytosed antigens as well as the MHC class II-associated invariant chain (Ii). Thus far, only the cysteine protease cathepsin (Cat) S appears essential for complete destruction of Ii. The enzymes involved in degradation of the antigens themselves remain to be identified. Degradation of antigens in vitro and experiments using protease inhibitors have suggested that Cat B and Cat D, two major aspartyl and cysteine proteases, respectively, are involved in antigen degradation. We have analyzed the antigen-presenting properties of cells derived from mice deficient in either Cat B or Cat D. Although the absence of these proteases provoked a modest shift in the efficiency of presentation of some antigenic determinants, the overall capacity of Cat B-/- or Cat D-/- antigen-presenting cells was unaffected. Degradation of Ii proceeded normally in Cat B-/- splenocytes, as it did in Cat D-/- cells. We conclude that neither Cat B nor Cat D are essential for MHC class II-mediated antigen presentation.

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Figures

Figure 1
Figure 1
Normal maturation of MHC class II in Cat B knock-out mice. Cat B+ (A) or Cat B− (B) splenocytes were metabolically labeled for 30 min and chased for the times indicated in the absence or presence of leupeptin (1 mM) or LHVS (3 nM). MHC class II molecules were immunoprecipitated with mAb N22 and run in 12.5% reducing SDS/PAGE without (NB) or after (B) boiling. The position of the class II α and β chains, Ii, the p41 form of Ii, SDS-stable (peptide-bound) αβ complexes, the leupeptin-induced polypeptide LIP10, and the I-Ab-LIP10 SDS-stable complex (21, 47, 48), are indicated
Figure 2
Figure 2
Antigen presentation by Cat D−/− APC. Splenocytes from Cat D+/− (solid bars) or Cat D−/− mice (shaded bars) were used as APC in assays of antigen recognition by the panel of hybridomas shown in Table 1 in the presence of the indicated concentration of antigen. The epitope recognized by each T cell line is shown under its name. The results shown are representative of at least two experiments in which all the hybridomas specific for the same MHC background were tested with the same combination of Cat D+/− and Cat D−/− splenocytes of the appropriate MHC specificity. All values are means of triplicate determinations, and the SD of each value is represented.
Figure 3
Figure 3
Antigen presentation by Cat B−/− APC. Splenocytes from Cat B+/+ (solid bars) or Cat B−/− mice (shaded bars) were used as APC in assays of antigen recognition by the hybridomas shown in the presence of the indicated concentration of antigen. The results shown are representative of two independent experiments in which each value is the mean of triplicate determinations.
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