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The death domain kinase RIP protects thymocytes from tumor necrosis factor receptor type 2-induced cell death.

Cusson N, Oikemus S, Kilpatrick ED, Cunningham L, Kelliher M - J. Exp. Med. (2002)

Bottom Line: Unlike Fas-associated death domain, rip does not regulate T cell proliferation, as rip-/- T cells respond to polyclonal activators.Surprisingly, the rip-associated thymocyte apoptosis was not rescued by the absence of TNFR1, but appears to be rescued by an absence of TNFR2.Taken together, this study implicates RIP and TNFR2 in thymocyte survival.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics and Microbiology, Program in Immunology/Virology, University of Massachusetts Medical School, Worcester, MA 01605, USA.

ABSTRACT
Fas and the tumor necrosis factor receptor (TNFR)1 regulate the programmed cell death of lymphocytes. The death domain kinase, receptor interacting protein (rip), is recruited to the TNFR1 upon receptor activation. In vitro, rip-/- fibroblasts are sensitive to TNF-induced cell death due to an impaired nuclear factor kappaB response. Because rip-/- mice die at birth, we were unable to examine the effects of a targeted rip mutation on lymphocyte survival. To address the contribution of RIP to immune homeostasis, we examined lethally irradiated mice reconstituted with rip-/- hematopoietic precursors. We observed a decrease in rip-/- thymocytes and T cells in both wild-type C57BL/6 and recombination activating gene 1-/- irradiated hosts. In contrast, the B cell and myeloid lineages are unaffected by the absence of rip. Thus, the death domain kinase rip is required for T cell development. Unlike Fas-associated death domain, rip does not regulate T cell proliferation, as rip-/- T cells respond to polyclonal activators. However, rip-deficient mice contain few viable CD4+ and CD8+ thymocytes, and rip-/- thymocytes are sensitive to TNF-induced cell death. Surprisingly, the rip-associated thymocyte apoptosis was not rescued by the absence of TNFR1, but appears to be rescued by an absence of TNFR2. Taken together, this study implicates RIP and TNFR2 in thymocyte survival.

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Related in: MedlinePlus

Flow cytometric analysis of mice reconstituted with rip+/+ or rip−/− fetal liver precursors. (A) Flow cytometric analysis of thymus. Single cell suspensions of thymocytes were stained for the donor-specific Ly9.1 marker and for CD4 or CD8 12 wk after reconstitution. (B) Flow cytometric analysis of peripheral lymphocytes. Single cell suspensions of cervical, inguinal, and mesenteric lymph nodes from rip+/+ and rip−/− reconstituted mice were stained with FITC-Ly9.1 and PE–anti-CD3, PE-CD4, or PE-CD8. (C) Splenocytes were also stained with FITC-Ly9.1 and CD3-PE, CD4-PE, CD8-PE, PE-B220, or PE–Mac-1. 10,000 events were collected. Plots are representative of three independent experiments.
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fig1: Flow cytometric analysis of mice reconstituted with rip+/+ or rip−/− fetal liver precursors. (A) Flow cytometric analysis of thymus. Single cell suspensions of thymocytes were stained for the donor-specific Ly9.1 marker and for CD4 or CD8 12 wk after reconstitution. (B) Flow cytometric analysis of peripheral lymphocytes. Single cell suspensions of cervical, inguinal, and mesenteric lymph nodes from rip+/+ and rip−/− reconstituted mice were stained with FITC-Ly9.1 and PE–anti-CD3, PE-CD4, or PE-CD8. (C) Splenocytes were also stained with FITC-Ly9.1 and CD3-PE, CD4-PE, CD8-PE, PE-B220, or PE–Mac-1. 10,000 events were collected. Plots are representative of three independent experiments.

Mentions: In addition to fewer cells in the rip−/− reconstituted thymus, the percentage of wild-type versus rip−/−-derived cells was also different. In rip+/+ reconstituted mice, 97% of the thymus contained Ly9.1+ cells, whereas only 3% of thymocytes in the rip−/− reconstituted mice were donor derived (Fig. 1 A). Although few rip−/− thymocytes were detected in the reconstituted mice, the CD4/CD8 profile did not reveal any developmental changes in rip−/− thymocytes.


The death domain kinase RIP protects thymocytes from tumor necrosis factor receptor type 2-induced cell death.

Cusson N, Oikemus S, Kilpatrick ED, Cunningham L, Kelliher M - J. Exp. Med. (2002)

Flow cytometric analysis of mice reconstituted with rip+/+ or rip−/− fetal liver precursors. (A) Flow cytometric analysis of thymus. Single cell suspensions of thymocytes were stained for the donor-specific Ly9.1 marker and for CD4 or CD8 12 wk after reconstitution. (B) Flow cytometric analysis of peripheral lymphocytes. Single cell suspensions of cervical, inguinal, and mesenteric lymph nodes from rip+/+ and rip−/− reconstituted mice were stained with FITC-Ly9.1 and PE–anti-CD3, PE-CD4, or PE-CD8. (C) Splenocytes were also stained with FITC-Ly9.1 and CD3-PE, CD4-PE, CD8-PE, PE-B220, or PE–Mac-1. 10,000 events were collected. Plots are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194008&req=5

fig1: Flow cytometric analysis of mice reconstituted with rip+/+ or rip−/− fetal liver precursors. (A) Flow cytometric analysis of thymus. Single cell suspensions of thymocytes were stained for the donor-specific Ly9.1 marker and for CD4 or CD8 12 wk after reconstitution. (B) Flow cytometric analysis of peripheral lymphocytes. Single cell suspensions of cervical, inguinal, and mesenteric lymph nodes from rip+/+ and rip−/− reconstituted mice were stained with FITC-Ly9.1 and PE–anti-CD3, PE-CD4, or PE-CD8. (C) Splenocytes were also stained with FITC-Ly9.1 and CD3-PE, CD4-PE, CD8-PE, PE-B220, or PE–Mac-1. 10,000 events were collected. Plots are representative of three independent experiments.
Mentions: In addition to fewer cells in the rip−/− reconstituted thymus, the percentage of wild-type versus rip−/−-derived cells was also different. In rip+/+ reconstituted mice, 97% of the thymus contained Ly9.1+ cells, whereas only 3% of thymocytes in the rip−/− reconstituted mice were donor derived (Fig. 1 A). Although few rip−/− thymocytes were detected in the reconstituted mice, the CD4/CD8 profile did not reveal any developmental changes in rip−/− thymocytes.

Bottom Line: Unlike Fas-associated death domain, rip does not regulate T cell proliferation, as rip-/- T cells respond to polyclonal activators.Surprisingly, the rip-associated thymocyte apoptosis was not rescued by the absence of TNFR1, but appears to be rescued by an absence of TNFR2.Taken together, this study implicates RIP and TNFR2 in thymocyte survival.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics and Microbiology, Program in Immunology/Virology, University of Massachusetts Medical School, Worcester, MA 01605, USA.

ABSTRACT
Fas and the tumor necrosis factor receptor (TNFR)1 regulate the programmed cell death of lymphocytes. The death domain kinase, receptor interacting protein (rip), is recruited to the TNFR1 upon receptor activation. In vitro, rip-/- fibroblasts are sensitive to TNF-induced cell death due to an impaired nuclear factor kappaB response. Because rip-/- mice die at birth, we were unable to examine the effects of a targeted rip mutation on lymphocyte survival. To address the contribution of RIP to immune homeostasis, we examined lethally irradiated mice reconstituted with rip-/- hematopoietic precursors. We observed a decrease in rip-/- thymocytes and T cells in both wild-type C57BL/6 and recombination activating gene 1-/- irradiated hosts. In contrast, the B cell and myeloid lineages are unaffected by the absence of rip. Thus, the death domain kinase rip is required for T cell development. Unlike Fas-associated death domain, rip does not regulate T cell proliferation, as rip-/- T cells respond to polyclonal activators. However, rip-deficient mice contain few viable CD4+ and CD8+ thymocytes, and rip-/- thymocytes are sensitive to TNF-induced cell death. Surprisingly, the rip-associated thymocyte apoptosis was not rescued by the absence of TNFR1, but appears to be rescued by an absence of TNFR2. Taken together, this study implicates RIP and TNFR2 in thymocyte survival.

Show MeSH
Related in: MedlinePlus