Single-cell analysis of signal transduction in CD4 T cells stimulated by antigen<i>in vivo</i>

Zell, Traci; Khoruts, Alexander; Ingulli, Elizabeth; Bonnevier, Jody; Mueller, Daniel L.; Jenkins, Marc K.

doi:10.1073/pnas.191567898

Cited by 72 publications

(64 citation statements)

References 35 publications

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“…A similar 2-fold inhibition was observed in vivo by intracellular staining following high dose peptide administration to the tolerant mice (data not shown; Ref. 62). The inhibition of ERK phosphorylation possibly stems from a decrease in RAS-GRP1 activity secondary to the impairment of PLC␥1 (63).…”

Section: Discussionsupporting

confidence: 78%

Adaptive Tolerance and Clonal Anergy Are Distinct Biochemical States

Chiodetti

Choi

Barber

et al. 2006

The Journal of Immunology

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Adaptive tolerance is a process by which T cells become desensitized when Ag stimulation persists following an initial immune response in vivo. To examine the biochemical changes in TCR signaling present in this state, we used a mouse model in which Rag2−/− TCR-transgenic CD4+ T cells were transferred into CD3ε−/− recipients expressing their cognate Ag. Compared with naive T cells, adaptively tolerant T cells had normal levels of TCR and slightly increased levels of CD4. Following activation with anti-TCR and anti-CD4 mAbs, the predominant signaling block in the tolerant cells was at the level of Zap70 kinase activity, which was decreased 75% in vitro. Phosphorylations of the Zap70 substrates (linker of activated T cells and phospholipase Cγ1 were also profoundly diminished. This proximal defect impacted mostly on the calcium/NFAT and NF-κB pathways, with only a modest decrease in ERK1/2 phosphorylation. This state was contrasted with T cell clonal anergy in which the RAS/MAPK pathway was preferentially impaired and there was much less inhibition of Zap70 kinase activity. Both hyporesponsive states manifested a block in IκB degradation. These results demonstrate that T cell adaptive tolerance and clonal anergy are distinct biochemical states, possibly providing T cells with two molecular mechanisms to curtail responsiveness in different biological circumstances.

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Section: Discussionsupporting

confidence: 78%

Adaptive Tolerance and Clonal Anergy Are Distinct Biochemical States

Chiodetti

Choi

Barber

et al. 2006

The Journal of Immunology

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“…[9][10][11][12][13][14] We describe here optimized conditions for p-ERK quantitation in primary patients cells by FCM (see Supplemental Information).…”

Section: Discussionmentioning

confidence: 99%

Quantitative single cell determination of ERK phosphorylation and regulation in relapsed and refractory primary acute myeloid leukemia

et al. 2005

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We investigated the constitutive activation of the MEK/ERK pathway in acute myelogenous leukemia (AML) via a flow cytometric technique to quantitate expression of phosphorylated ERK (p-ERK). A total of 42 AML samples (16 newly diagnosed, 26 relapsed/refractory) were analyzed. Normal bone marrow CD34(+) cells (n = 10) had little or no expression of p-ERK, while G-CSF-mobilized CD34(+) cells exhibited enhanced p-ERK levels. Markedly elevated p- ERK levels were found in 83.3% of the AML samples, with no differences observed between the newly diagnosed and relapsed/refractory samples. Treatment with a MEK inhibitor resulted in significantly decreased p- ERK levels in both the newly diagnosed and relapsed/refractory samples, which was associated with growth arrest, but not apoptosis induction. In summary, we defined conditions for the analysis of MAPK signaling in primary AML samples. Normal CD34(+) cells expressed very low levels of p- ERK, and increased p- ERK levels were found in normal G-CSF-stimulated circulating CD34(+) cells. Constitutively high p-ERK levels observed in the majority of AML samples suggest deregulation of this pathway that appears to be independent of disease status. The ability of ERK inhibition to promote growth arrest rather than apoptosis suggests that clinical trials of MEK/ERK inhibitors may be more effective when combined with chemotherapy

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“…into live naive P14 mice, or into P14 effector or memory mice. We sacrificed them 15 min later, and immediately fixed splenocytes to preserve activated molecules (37). We then analyzed phospho-LAT by intracellular FACS analysis using an Ab to phospho-LAT (tyrosine 191).…”

Section: More Lipid Rafts With More Phosphoproteins In Effector and Mmentioning

confidence: 99%

TCR Signal Transduction in Antigen-Specific Memory CD8 T Cells

Kersh

Kaech

Onami

et al. 2003

The Journal of Immunology

102

101

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Memory T cells are more responsive to Ag than naive cells. To determine whether memory T cells also have more efficient TCR signaling, we compared naive, effector, and memory CD8 T cells of the same antigenic specificity. Surprisingly, initial CD3 signaling events are indistinguishable. However, memory T cells have more extensive lipid rafts with higher phosphoprotein content before TCR engagement. Upon activation in vivo, they more efficiently induce phosphorylation of-LAT (linker for activation of T cells), ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase), and p38. Thus, memory CD8 T cells do not increase their TCR sensitivity, but are better poised to augment downstream signals. We propose that this regulatory mechanism might increase signal transduction in memory T cells, while limiting TCR cross-reactivity and autoimmunity.

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Single-cell analysis of signal transduction in CD4 T cells stimulated by antigenin vivo

Cited by 72 publications

References 35 publications

Adaptive Tolerance and Clonal Anergy Are Distinct Biochemical States

Adaptive Tolerance and Clonal Anergy Are Distinct Biochemical States

Quantitative single cell determination of ERK phosphorylation and regulation in relapsed and refractory primary acute myeloid leukemia

TCR Signal Transduction in Antigen-Specific Memory CD8 T Cells

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