Cellular and peptide requirements for in vitro clonal deletion of immature thymocytes

K. Iwabuchi, K. I. Nakayama, R. L. McCoy, F. Wang, T. Nishimura, S. Habu, K. M. Murphy, D. Y. Loh

Research output: Contribution to journalArticle

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Abstract

Thymocytes from DO10 T-cell-receptor transgenic mice undergo apoptosis, or programmed cell death, when chicken ovalbumin-(323-339) peptide is administered in vivo. Using DO10 mice thymocytes, we have now developed a simple in vitro model system that recapitulates the in vivo clonal-deletion process. When transgenic thymocytes were cocultured with fibroblasts, B cells, or thymic nurse cell lines (all bearing I-A(d)) in the presence of chicken ovalbumin-(323-339), deletion of the transgenic TCR+CD4+CD8+ thymocytes was seen within 8-20 hr. Thymocytes designed to bear I-A(d) on their surface could mediate the deletion themselves. Thus, thymocyte clonal deletion entirely depends on the stage at which the thymocytes are vulnerable to the onset of apoptosis, rather than on the nature of the peptide antigen- presenting cells. Furthermore, thymic nurse cell line TNC-R3.1 could cause deletion, strongly suggesting that some thymic epithelial/stromal components are potentially capable of participating in negative selection. In all cases examined, little deletion could be induced at a peptide concentration <10 nM, thus defining the minimum amount of peptide antigen required for negative selection. The peptide-dependent in vitro negative-selection system will allow further dissection of the molecular and cellular processes involved in clonal deletion due to apoptosis in the thymus.

Original languageEnglish
Pages (from-to)9000-9004
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number19
DOIs
Publication statusPublished - Oct 13 1992
Externally publishedYes

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Clonal Deletion
Thymocytes
Peptides
Apoptosis
Chickens
Nurses
Cell Line
Ovalbumin
Antigen-Presenting Cells
T-Cell Antigen Receptor
In Vitro Techniques
Thymus Gland
Transgenic Mice
Dissection
B-Lymphocytes
Cell Death
Fibroblasts
Antigens

All Science Journal Classification (ASJC) codes

  • General

Cite this

Cellular and peptide requirements for in vitro clonal deletion of immature thymocytes. / Iwabuchi, K.; Nakayama, K. I.; McCoy, R. L.; Wang, F.; Nishimura, T.; Habu, S.; Murphy, K. M.; Loh, D. Y.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 19, 13.10.1992, p. 9000-9004.

Research output: Contribution to journalArticle

Iwabuchi, K. ; Nakayama, K. I. ; McCoy, R. L. ; Wang, F. ; Nishimura, T. ; Habu, S. ; Murphy, K. M. ; Loh, D. Y. / Cellular and peptide requirements for in vitro clonal deletion of immature thymocytes. In: Proceedings of the National Academy of Sciences of the United States of America. 1992 ; Vol. 89, No. 19. pp. 9000-9004.
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