ABSTRACT
Mouse thymic stromal cell line 4 (MTSC4) is one of the stromal cell lines established in our laboratory. While losing the characteristics of epithelial cells, they express some surface markers shared with thymic dendritic cells (TDCs). To further study the biological functions of these cells, we compared the capability of MTSC4 with TDCs in the induction of thymocyte apoptosis, using thymic reaggregation culture system. Apoptosis of thymocytes induced by MTSC4 and TDCs was measured by Annexin V and PI staining and analyzed by flow cytometry. We found that MTSC4 selectively augmented the apoptosis of CD4+8+ (DP) thymocytes. This effect was Fas/FasL independent and could not be blocked by antibodies to MHC class I and class II molecules. In addition, MTSC4 enhanced the apoptosis of DP thymocytes from different strains of mice, which implies that MTSC4-induced thymocyte apoptosis is not mediated by the TCR recognition of self peptide/MHC molecules. In contrast to MTSC4, thymocyte apoptosis induced by TDCs was MHC-restricted. Thus, MHC-independent fashion of stromal-DP thymocyte interaction may be one of the ways to induce thymocyte apoptosis in thymus. Our study has also shown that the interaction of MTSC4 stromal cells and thymocytes is required for the induction of thymocyte apoptosis.
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Abbreviations
- BSS:
-
balanced salt solution
- DN:
-
double-negative
- DP:
-
double-positive
- Int:
-
intermediate
- MTEC1:
-
mouse thymic epithelial cell line 1
- MTSC4:
-
mouse thymic stromal cell line 4
- NCS:
-
newborn calf serum
- SP:
-
single-positive
- TDC:
-
thymic dendritic cell
- TRC:
-
thymic reaggregation culture
References
Anderson G, Moore NC, Owen JJT, Jenkinson EJ . Cellular interactions in thymocyte development. Annu Rev Immunol 1996; 14:73–99.
Sebzda E, Mariathasan S, Ohteki T, Jones R, Bachmann MF, Ohashi PS . Selection of the T cell repertoire. Annu Rev Immunol 1999; 17:829–74.
Brocker T, Riedinger M, Karjalainen K . Targeted expression of major histocompatibility complex (MHC) class II molecules demonstrates that dendritic cells can induce negative but not positive selection of thymocytes in vivo. J Exp Med 1997; 185(3):541–50.
Anderson G, Partington KM, Jenkinson EJ . Differential effects of peptide diversity and stromal cell type in positive and negative selection in the thymus. J Immunol 1998; 161:6599–603.
Pircher H ., K. Brduscha, U. Steinhoff, et al. Tolerance induction by clonal deletion of CD4+CD8+ thymocytes in vitro does dot require dedicated antigen-presenting cells. Eur J Immunol 1993; 23:669–74.
Tanaka Y, Mamalaki C, Stockinger B, Kioussis D . In vitro negative selection of αβ T cell receptor transgenic thymocytes by conditionally immortalized thymic cortical epithelial cell lines and dendritic cells. Eur J Immunol 1993; 23:2614–21.
Capone M, Romagnoli P, Beermann F, MacDonald HR, van Meerwijk JPM . Dissociation of thymic positive and negative selection in transgenic mice expressing major histocompatibility complex class I molecules exclusively on thymic cortical epithelial cells. Blood 2001; 97:1336–42.
Shu SZ, Chen WF, Liu ZG, Zhang PX, Wu JS . Establishment and identification of eight mouse thymic stromal cell lines. J. Beijing Medical University. (in Chinese) 1993; 25:237–41.
Pei LH, Liu ZG, Chen WF . Manipulation of mouse dendritic cell line clones and their characteristics. Chin J Immunol (in Chinese) 1999; 15:257–9.
Vremec D, Zorbas M, Scollay R, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: Investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med 1992; 176:47–58.
Lu LS, Chen WF . The functional development and the expression of Qa-2 antigen onthe medullary CD4+CD8− cells by the MTEC1 cells. Science in China (in Chinese) Ser B 1995; 25:1161–6.
Dong HD, Pang XW, Chen WF . The different effects of two thymic stromal cells on programmed cell death of thymocytes cultured in vitro. Chin J Microbiol Immunol (in Chinese) 1996; 16:284–8.
Klein L, Kyewski B . Self-antigen presentation by thymic stromal cells: subtle division of labor. Curr Opin in Immunol 2000; 12:179–86.
Sebzda E, Wallace VA, Mayer J, eung RSMY, Mak TW, Ohashi PS . Positive and negative thymocyte selection induced by different concentrations of a single peptide. Science 1994; 263:1615–8.
Delaney JR, Sykulev Y, Eisen HN, Tonegawa S . Differences in the level of expression of class I major histocompatibility complex proteins on thymic epithelial and dendritic cells influence the decision of immature thymocytes between positive and negative selection. Proc Natl Acad Sci USA 1998; 95:5235–40.
Dong HD, Chen J, Chen WF . Membrane molecules in induction of apoptosis of thymocytes by mouse thymic dendritic cells which express Fas ligands. Science in China (Series C) 1998; 41:189–94.
Liu ZG, Chen WF, Cao LX, Pang XW, Zhang PX, Yu Q . Cytokine production by MTSC4 cells, an established mouse thymic dendritic cell line. Acta Biologiae Experimentalis Sinica (in Chinese) 1994; 27:91–8.
Pang XW, Wu Q, Qian XP, Wu JS, Chen WF . Capability of antigen presentation by mouse thymic stromal cells. Chin J Microbiol Immunol (in Chinese) 2000; 20:28–31.
Surh CD, Sprent J . T-cell apoptosis detected in situ during positive and negative selection in the thymus. Nature 1994; 372:100–3.
Guo J, Hawwari A, Li H, Sun Z, Mahanta SK . Regulation of the TCRalpha repertoire by the survival window of CD4+8+ thymocytes. Nat Immunol 2002; 3:469–76.
Amsen,D . and A.M.Kruisbeek . Thymocyte selection: not by TCR alone. Immunological Reviews 1998; 165:209–29.
Lenschow DJ, Walunas TL, Bluestone JA . CD28/B7 system of T cell costimulation. Annu Rev Immunol 1996; 14:233–58.
Foy TM, Page DM, Waldschmidt TJ . et al. An essential role for gp39, the ligand for CD40, in thymic selection. J Exp Med 1995; 182:1377–88.
Kishimoto H, Surh CD, Sprent J . A role for Fas in negative selection of thymocytes in vivo. J Exp Med 1998; 187:1427–38.
Acknowledgements
The authors would like to thank Jia Ping TAO and Shi Liang MA for FACS sorting.
This work was funded by grants from Major State Bascic Research Development Program of China (No. G1999053904), National Natural Science Foundation of China (No. 30271200) and Young Scientist Cooperation Research Fund (No. 39928019).
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HE, X., LI, J., QIAN, X. et al. The thymic stromal cell line MTSC4 induced thymocyte apoptosis in a non-MHC-restricted manner. Cell Res 14, 125–133 (2004). https://doi.org/10.1038/sj.cr.7290211
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DOI: https://doi.org/10.1038/sj.cr.7290211