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Perturbation of the T Lymphocyte Lineage In Transgenic Mice Expressing a Constitutive Repressor of Nuclear Factor (NF)-B

Mark R. Boothby^*,, Ana L. Mora^*,, David C. Scherer^*,, Jeffrey A. Brockman^*,, and Dean W. Ballard^*,

From the ^* Department of Microbiology and Immunology; Rheumatology Division, Department of Medicine; and the Howard Hughes Medical Institute, Vanderbilt University Medical Center, Nashville, Tennessee 37232

Members of the nuclear factor (NF)-B/Rel family transcription factors are induced during thymic selection and in mature T lymphocytes after ligation of the T cell antigen receptor (TCR). Despite these findings, disruption of individual NF-B/Rel genes has revealed no intrinsic defect in the development of mature T cells, perhaps reflecting functional redundancy. To circumvent this possibility, the T cell lineage was targeted to express a trans-dominant form of IB that constitutively represses the activity of multiple NF-B/Rel proteins. Transgenic cells expressing this inhibitor exhibit a significant proliferative defect, which is not reversed by the addition of exogenous interleukin-2. Moreover, mitogenic stimulation of splenocytes leads to increased apoptosis of transgenic T cells as compared with controls. In addition to deregulated T cell growth and survival, transgene expression impairs the development of normal T cell populations as evidenced by diminished numbers of TCR^hi CD8 single-positive thymocytes. This defect was significantly amplified in the periphery and was accompanied by a decrease in CD4⁺ T cells. Taken together, these in vivo findings indicate that the NF-B/Rel signaling pathway contains compensatory components that are essential for the establishment of normal T cell subsets.

The authors acknowledge A. Cherrington and W. Armistead for technical assistance; J. Wright and C. Pettipher (Vanderbilt Cancer Center Transgenic Core) for DNA microinjections; A. Lackey (Cytometry Associates, Brentwood, TN) and J. Price (Vanderbilt Cancer Center Core) for expert flow cytometry analyses; L. Glimcher, E. Oltz, T. Aune, G. Miller, D. Perkins, T. Laufer, J. Chen, L. Van Kaer, E. Robey, D. Kioussis, L. Kelley, and P. Fink for helpful discussions; and E. Vance for help with manuscript preparation. D.W. Ballard is an investigator of the Howard Hughes Medical Institute (HHMI). Supported by National Institutes of Health grant AI-33839 and the Howard Hughes Medical Institute (D.C. Scherer, J.A. Brockman, D.W. Ballard), and by National Institutes of Health grant AI-36997, GM-42550, and the Leukemia Society of America (A.L. Mora, M.R. Boothby).

¹Abbreviations used in this paper: BrdU, 5-bromo-2'-deoxyuridine; B-pd, B enhancer sequences based on the IL-2R promoter; NF-B, nuclear factor-B; SP, single positive.

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