Daysianne de Lira - Academia.edu (original) (raw)

Immunity, 1999

Cathepsins have been implicated in the degradation Rodriguez and Diment, 1995; Riese et al., 1996... more Cathepsins have been implicated in the degradation Rodriguez and Diment, 1995; Riese et al., 1996). Howof proteins destined for the MHC class II processing ever, in sharp contrast with many biochemical in vitro pathway and in the proteolytic removal of invariant experiments, genetic analyses using cathepsin B-and chain (Ii), a critical regulator of MHC class II function. D-deficient mice revealed that these two major lyso-Mice lacking the lysosomal cysteine proteinase casomal proteinases are not necessary for Ii degradation thepsin S (catS) demonstrated a profound inhibition and processing of a number of exogenous and endogeof Ii degradation in professional APC in vivo. A marked nous protein antigens (Villadangos et al., 1997; Deussing variation in the generation of MHC class II-bound Ii et al., 1998; T. N. et al., unpublished data). This sugfragments and presentation of exogenous proteins gested that the complexity and redundancy of lysowas observed between B cells, dendritic cells, and somal proteinases may impede the elucidation of the macrophages lacking catS. CatS-deficient mice showed function of individual enzymes. However, our recent diminished susceptibility to collagen-induced arthritis, analysis of cathepsin L (catL)-deficient mice revealed suggesting a potential therapeutic target for regulaa profound defect in Ii degradation in thymic cortical tion of immune responsiveness. epithelial cells (cTEC) but not in bone marrow-derived APCs (B cells, DC, and Mph). This tissue-specific defect correlates with the restricted pattern of expression of later. Macrophages were cultured in teflon plates in the presence of 50 U/ml recombinant IFN␥ for 48 hr. Nonadherent cells were We thank M. Appleby, F. Ramsdell, and D. Walker (Chiroscience, washed with warm medium, and macrophages were harvested by Bothell, WA) for generous assistance with cell sorting; Immunex trypsinization. The class II expression on activated Mph was as-Corporation for providing recombinant human Flt3 ligand; D. Wilson sessed by staining in the presence of Fc receptor blocking MAb and M. Gomez for technical assistance; H. Caldwell (NIAID Rocky 24G.2 (ATCC 197) with F4/80-FITC MAb (Pharmingen) and biotinyl-Mountain Laboratory) for C. trachomatis and 116.3 T hybrids; S. ated anti-class II MAbs followed by streptavidin-Tricolor. Purity of Kahn (University of Washington) for recombinant SA85 T. cruzi prothe macrophage population was over 95% as determined by two tein and 71.5 T hybrids; N. Shastri (UC Berkeley) for BO4 T hybrids; color flow cytometry. M. Bevan, P. Fink, and P. Wong for reading of the manuscript; and members of our laboratories for helpful comments and discussion. . Kovats et al., 1998) were cultured in duplicates for 20 hr with variable numbers of control or mutant SC. In parallel, 1 ϫ 10 5 SC were References incubated with I-A b -restricted T hybrids (Nakagawa et al., 1998) specific for HEL (BO4), KLH (2BH11. A1), myoglobin (1BE6A1), T. Avva, R.R., and Cresswell, P. (1994). In vivo and in vitro formation cruzi SA85 (71.5), and C. trachomatis (116.3) and I-A q -restricted T and dissociation of HLA-DR complexes with invariant chain-derived cell hybrids specific for HEL (qLy30.0), OVA (qO/H166.0), and bovine peptides. Immunity 1, 763-774. collagen type II (qCII85.33) (Rosloniec et al., 1996) with variable amounts of exogenous antigens. IL-2 production was assessed us-Barrett, A.J. (1987). The cystatins: a new class of peptidase inhibiing HT-2 proliferation as determined by the Alamar Blue colorimetric tors. Trends Biochem. Sci. 12, 193-196. assay. The results are expressed as arbitrary OD units (A570-A600). Baumgart, M., Moos, V., Schuhbauer, D., and Muller, B. (1998). Differential expression of major histocompatibility complex class II Induction and Scoring of Collagen-Induced Arthritis genes on murine macrophages associated with T cell cytokine pro-CIA was induced and scored in male catS Ϫ/Ϫ or wt mice (9-13 weeks file and protective/suppressive effects. Proc. Natl. Acad. Sci., USA old; 10-12 animals per group) immunized at the base of the tail on 95, 6936-6940. day 0 and 21 with chick type II collagen in CFA as described (Griffiths Bennett, K., Levine, T., Ellis, J.S., Peanasky, R.J., Samloff, I.M., Kay, et al., 1995). Severity of the disease was evaluated by inspection of J., and Chain, B.M. (1992). Antigen processing for presentation by the paws and joints and a score was assigned (0, normal paw; 1, class II major histocompatibility complex requires cleavage by caswelling and/or redness of one toe or finger joint; 2, two or more thepsin E. Eur. J. Immunol. 22, 1519-1524. joints involved; 3, severe arthritis in the entire paw; maximum score Bevec, T., Stoka, V., Pungercic, G., Dolenc, I., and Turk, V. (1996). for each animal-12). Collagen-specific antibodies were measured Major histocompatibility complex class II-associated p41 invariant in the serum of mice at the termination of the experiment (day 58) by ELISA. Immune and control sera were titrated on Nunc-Immuno chain fragment is a strong inhibitor of lysosomal cathepsin L. J. Maxisorp plates coated with chick collagen (Marie Griffith, University Exp. Med. 183, 1331-1338. of Utah). The presence of collagen-specific antibodies was deter-Blum, J.S., and Cresswell, P. (1988). Role of intracellular proteases mined with goat anti-mouse IgG1, IgG2a, IgG2b, and IgG3 HRPin the processing and transport of class II HLA antigens. Proc. Natl. conjugated antibodies (Southern Biotech). Total collagen-specific Acad. Sci., USA 85, 3975-3980. IgG and IgM was measured with analogous HRP-conjugated goat Brachet, V., Raposo, G., Amigorena, S., and Mellman, I. (1997). Ii antibodies (Jackson ImmunoResearch Laboratory, Inc.). chain controls the transport of major histocompatibility complex class II molecules to and from lysosomes. . (1994). Characterization of the T cell Proteinase Active Site Labeling determinants in the induction of autoimmune arthritis by bovine Splenocytes, DC, B cells, and Mph were preincubated for 1 hr at 37ЊC in methionine/cysteine-free RPMI 1640 supplemented with 200 ␣1(II)-CB11 in H-2 q Mice. J. Immunol. 152, 3088-3097.