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. 1997 Jun 10;94(12):6216-21.
doi: 10.1073/pnas.94.12.6216.

A new member of the tumor necrosis factor/nerve growth factor receptor family inhibits T cell receptor-induced apoptosis

G Nocentini  1 , L Giunchi, S Ronchetti, L T Krausz, A Bartoli, R Moraca, G Migliorati, C Riccardi
Affiliations

A new member of the tumor necrosis factor/nerve growth factor receptor family inhibits T cell receptor-induced apoptosis

G Nocentini et al. Proc Natl Acad Sci U S A. .

Abstract

By comparing untreated and dexamethasone-treated murine T cell hybridoma (3DO) cells by the differential display technique, we have cloned a new gene, GITR (glucocorticoid-induced tumor necrosis factor receptor family-related gene) encoding a new member of the tumor necrosis factor/nerve growth factor receptor family. GITR is a 228-amino acids type I transmembrane protein characterized by three cysteine pseudorepeats in the extracellular domain and similar to CD27 and 4-1BB in the intracellular domain. GITR resulted to be expressed in normal T lymphocytes from thymus, spleen, and lymph nodes, although no expression was detected in other nonlymphoid tissues, including brain, kidney, and liver. Furthermore, GITR expression was induced in T lymphocytes upon activation by anti-CD3 mAb, Con A, or phorbol 12-myristate 13-acetate plus Ca-ionophore treatment. The constitutive expression of a transfected GITR gene induced resistance to anti-CD3 mAb-induced apoptosis, whereas antisense GITR mRNA expression lead to increased sensitivity. The protection toward T cell receptor-induced apoptosis was specific, because other apoptotic signals (Fas triggering, dexamethasone treatment, or UV irradiation) were not modulated by GITR transfection. Thus, GITR is a new member of tumor necrosis factor/nerve growth factor receptor family involved in the regulation of T cell receptor-mediated cell death.

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Figures

Figure 1
Figure 1
Modulation of GITR mRNA of 3DO cells after DEX treatment (10−7 M), analyzed by Northern blotting (A) and competitive RT-PCR (B). (A) A β-actin probe was hybridized to the same filter as the internal control. (B) The expected length of the PCR products were 120 bp for GITR and 180 bp for the competitor. (C) Modulation of GITR gene expression investigated by run-on assay. The densitometric analysis of the film is reported in the histogram: after normalization with β-actin, a significant (P < 0.01) 6-fold increase was observed in the 24-h DEX-treated cells.
Figure 2
Figure 2
Protein putatively coded by GITR. Potential glycosylation (∗) and phosphorylation (#) sites are reported. The cysteine pseudorepeats and the respective cysteine position into the repeats (from 1 to 6 for cysteines from the first to the sixth position, and x for the extra cysteines, as referred to the canonical repeat) are also reported.
Figure 3
Figure 3
(A) Homology among the GITR cysteine pseudorepeats and those of the other murine members of TNF/NGFR family (the nontruncated only). After the name of the protein, the position of the pseudorepeat is reported (with respect to the other pseudorepeats of the protein and with respect to the position of the residues in the native protein giving the pseudorepeat). Amino acids identical or with similar function (R, K, H; D, E, N, Q; V, L, I, M; S, T; A, G) in more than 50% of the proteins were considered to be consensus and shaded; the identical are in boldface type. The respective cysteine positions in the repeats are also reported (see Fig. 2). (B) Homology among the cytoplasmic domains of murine and human CD27 and 4–1BB and of GITR. Charge (∧) of the amino acid residues present in at least two chains belonging to different receptors is indicated.
Figure 4
Figure 4
Expression of GITR in tissues analyzed by Northern blotting (A) and competitive RT-PCR (B). Overexpression of GITR in lymphocytes from thymus (C), spleen (D), and lymph nodes (E) activated with different stimuli for 1 and 3 days. The expected length of the PCR products were 120 bp for GITR and 180 bp for the competitor.
Figure 5
Figure 5
Expression of exogenous GITR (A) and antisense GITR (RTIG) (B) in transfected 3DO cells as evaluated by competitive RT-PCR. The expected length of the PCR products was 1074 bp for GITR and RTIG and 851 bp for the competitor. (C) Anti-CD3-induced apoptosis of GITR or RTIG overexpressing clones in comparison to apoptosis of untransfected (nuc) and pCR3-transfected clones. The expression of TCR–CD3 was similar in transfected and untransfected clones (data not shown).
Figure 6
Figure 6
Fas- (A), DEX- (B), and UV- (C) induced apoptosis of GITR or RTIG overexpressing clones in comparison to apoptosis of untransfected (nuc) and pCR3-transfected clones. Sensitivity of overexpressing clones was not significantly different from that of control clones (P > 0.05).
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References

    1. Wyllie A H, Kerr J F, Currie A R. Int Rev Cytol. 1980;68:251–306. - PubMed
    1. Smith C A, Farrah T, Goodwin R G. Cell. 1994;76:959–962. - PubMed
    1. Beutler B, van Huffel C. Science. 1994;264:667–668. - PubMed
    1. Flier J S, Underhill L H. N Engl J Med. 1996;334:1717–1725. - PubMed
    1. Gruss H, J, Dower S K. Blood. 1995;85:3378–3404. - PubMed

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