Critical requirement for the membrane-proximal cytosolic tyrosine residue for CD28-mediated costimulation in vivo

Y. Harada, M. Tokushima, Y. Matsumoto, S. Ogawa, M. Otsuka, Katsuhiko Hayashi, B. D. Weiss, C. H. June, R. Abe

Research output: Contribution to journalArticle

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Abstract

The YMNM motif that exists in the CD28 cytoplasmic domain is known as a binding site for phosphatidylinositol 3-kinase and Grb-2 and is considered to be important for CD28-mediated costimulation. To address the role of the YMNM motif in CD28 cosignaling in primary T cells, we generated transgenic mice on a CD28 null background that express a CD28 mutant lacking binding ability to phosphatidylinositol 3-kinase and Grb-2. After anti-CD3 and anti-CD28 Ab stimulation in vitro, the initial proliferative response and IL-2 secretion in CD28 Y189F transgenic T cells were severely compromised, while later responses were intact. In contrast to anti-CD3 and anti-CD28 Ab stimulation, PMA and anti-CD28 Ab stimulation failed to induce IL-2 production from CD28 Y189F transgenic T cells at any time point. Using the graft-vs-host reaction system, we assessed the role of the YMNM motif for CD28-mediated costimulation in vivo and found that CD28 Y189F transgenic spleen cells failed to engraft and could not induce acute graft-vs-host reaction. Together, these results suggest that the membrane-proximal tyrosine of CD28 is required for costimulation in vivo. Furthermore, these results indicate that the results from in vitro assays of CD28-mediated costimulation may not always correlate with T cell activation in vivo.

Original languageEnglish
Pages (from-to)3797-3803
Number of pages7
JournalJournal of Immunology
Volume166
Issue number6
DOIs
Publication statusPublished - Mar 15 2001

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Tyrosine
Graft vs Host Reaction
Phosphatidylinositol 3-Kinase
T-Lymphocytes
Membranes
Interleukin-2
Transgenic Mice
Spleen
Binding Sites
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Critical requirement for the membrane-proximal cytosolic tyrosine residue for CD28-mediated costimulation in vivo. / Harada, Y.; Tokushima, M.; Matsumoto, Y.; Ogawa, S.; Otsuka, M.; Hayashi, Katsuhiko; Weiss, B. D.; June, C. H.; Abe, R.

In: Journal of Immunology, Vol. 166, No. 6, 15.03.2001, p. 3797-3803.

Research output: Contribution to journalArticle

Harada, Y, Tokushima, M, Matsumoto, Y, Ogawa, S, Otsuka, M, Hayashi, K, Weiss, BD, June, CH & Abe, R 2001, 'Critical requirement for the membrane-proximal cytosolic tyrosine residue for CD28-mediated costimulation in vivo', Journal of Immunology, vol. 166, no. 6, pp. 3797-3803. https://doi.org/10.4049/jimmunol.166.6.3797
Harada, Y. ; Tokushima, M. ; Matsumoto, Y. ; Ogawa, S. ; Otsuka, M. ; Hayashi, Katsuhiko ; Weiss, B. D. ; June, C. H. ; Abe, R. / Critical requirement for the membrane-proximal cytosolic tyrosine residue for CD28-mediated costimulation in vivo. In: Journal of Immunology. 2001 ; Vol. 166, No. 6. pp. 3797-3803.
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