Gads−/− Mice Reveal Functionally Distinct Subsets of TCRβ+ CD4−CD8− Double-Negative Thymocytes

Zeng, Ling; Dalheimer, Stacy L.; Yankee, Thomas M.

doi:10.4049/jimmunol.179.2.1013

Cited by 14 publications

(50 citation statements)

References 37 publications

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Section: Discussionmentioning

confidence: 99%

“…In Gads -/-mice and in transgenic mice expressing an SH2 domain deletion of Gads, there is a decrease in the number of SP thymocytes [46][47][48]. Additionally, Gads -/-mice have an increase in the percentage of DN thymocytes and defects in survival and proliferation during the transition from the DN3 to DP stage [49].Consistent with these data, we observed an increase in the relative proportion of DN3 cells and a substantial effect on SP thymocyte development in GBF-expressing mice. However, we did not observe a specific defect in the relative proportion of DP thymocytes using the retroviral system.…”

mentioning

confidence: 99%

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In vivo disruption of T cell development by expression of a dominant‐negative polypeptide designed to abolish the SLP‐76/Gads interaction

Jordan¹,

Kliche²,

Shabason³

et al. 2007

Eur J Immunol

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Multi-molecular complexes nucleated by adaptor proteins play a central role in signal transduction. In T cells, one central axis consists of the assembly of several signaling proteins linked together by the adaptors linker of activated T cells (LAT), Src homology 2 domain-containing leukocyte-specific phosphoprotein of 76 kDa (SLP-76), and Grb2-related adaptor downstream of Shc (Gads). Each of these adaptors has been shown to be important for normal T cell development, and their proper sub-cellular localization is critical for optimal function in cell lines. We previously demonstrated in Jurkat T cells and a rat basophilic leukemic cell line that expression of a 50-amino acid polypeptide identical to the site on SLP-76 that binds to Gads blocks proper localization of SLP-76 and SLP-76-dependent signaling events. Here we extend these studies to investigate the ability of this polypeptide to inhibit TCR-induced integrin activity in Jurkat cells and to inhibit in vivo thymocyte development and primary T cell function. These data provide evidence for the in vivo function of a dominant-negative peptide based upon the biology of SLP-76 action and suggest the possibility of therapeutic potential of targeting the SLP-76/Gads interaction. IntroductionStimulation of T cells through the T cell receptor (TCR) leads to recruitment and localization of several critical signaling proteins, which results in the generation of a macro-molecular signaling complex. Key components of this complex, or signalosome, include the adaptors Src homology 2 (SH2) domain-containing leukocyte-specific phosphoprotein of 76 kDa (SLP-76), and Grb2-related adaptor downstream of Shc (Gads), which serves to link SLP-76 and its associated molecules to the transmembrane adaptor linker of activated T cells (LAT) [1].As an adaptor protein, SLP-76 possesses no enzymatic activity but functions via interaction with multiple proteins by way of three structural domains [2][3][4]. The N terminus harbors three tyrosines that become phos- phorylated upon TCR engagement [5,6] and have been reported to serve as docking sites for the guanine nucleotide exchange factor Vav, the adaptor protein Nck, the Tec family kinase Itk, and the p85 subunit of PI3K [7][8][9][10][11][12][13][14][15][16]. A single SH2 domain resides in the C-terminal portion of SLP-76 and links it to the adhesion and degranulation-promoting adaptor protein (ADAP) and to the hematopoietic progenitor kinase 1 [17][18][19]. Hematopoietic progenitor kinase 1, in turn, has been shown to phosphorylate SLP-76, creating a binding site for 14-3-3, a negative regulator of TCR signaling [20,21]. The central proline-rich region binds phospholipase Cc- Targeted deletion of SLP-76 in mice results in an inability of thymocytes to progress past the CD4 -CD8 -double-negative (DN) stage of development and thus a complete absence of mature T cells [28,29]. Since T cells do not develop in the absence of SLP-76, much of our knowledge of the role of SLP-76 in mature TCR signaling has been derived from the study of J14 ...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

In vivo disruption of T cell development by expression of a dominant‐negative polypeptide designed to abolish the SLP‐76/Gads interaction

Jordan¹,

Kliche²,

Shabason³

et al. 2007

Eur J Immunol

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“…Lck-Cre-dependent cleavage is typically thought to occur in CD4/CD8 double-negative stage 2 (DN2; CD25 ϩ /CD44 ϩ ) to double-negative stage 4 (DN4; CD25 Ϫ /CD44 Ϫ ), and small numbers of GFP ϩ cells were first detected in DN2, but they continued to accumulate through DN3 (CD25 Ϫ /CD44 ϩ ) and into DN4 (data not shown) (30,31). When we gated on GFP ϩ cells, we noted 50% more CD25…”

Section: Hdac3 Is Essential For Efficient Development Of Functional Tmentioning

confidence: 99%

Histone Deacetylase 3 Is Required for Efficient T Cell Development

Stengel

Zhao

Klus

et al. 2015

Molecular and Cellular Biology

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Hdac3 is a key target for Hdac inhibitors that are efficacious in cutaneous T cell lymphoma. Moreover, the regulation of chromatin structure is critical as thymocytes transition from an immature cell with open chromatin to a mature T cell with tightly condensed chromatin. To define the phenotypes controlled by Hdac3 during T cell development, we conditionally deleted Hdac3 using the Lck-Cre transgene. This strategy inactivated Hdac3 in the double-negative stages of thymocyte development and caused a significant impairment at the CD8 immature single-positive (ISP) stage and the CD4/CD8 double-positive stage, with few mature CD4؉ or CD8 ؉ single-positive cells being produced. When Hdac3 ؊/؊ mice were crossed with Bcl-xL-, Bcl2-, or TCR␤-expressing transgenic mice, a modest level of complementation was found. However, when the null mice were crossed with mice expressing a fully rearranged T cell receptor ␣␤ transgene, normal levels of CD4 single-positive cells were produced. Thus, Hdac3 is required for the efficient transit from double-negative stage 4 through positive selection.

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“…Cells were filtered through a 100 mm nylon mesh. Surface, intracellular staining and DNA staining were performed as described earlier (Zeng et al, 2007). Briefly, cells were surface labelled by incubating cells with antibodies for 30 min on ice in staining buffer [PBS containing 2% Fetal Clone I (HyClone Laboratories, Inc., Logan, UT, USA)].…”

Section: Cell Labelling and Flow Cytometrymentioning

confidence: 99%

“…DN thymocytes can be divided into the DN1 (CD44 hi CD25 -), DN2 (CD44 hi CD25 + ), DN3E (CD44 lo CD25 hi ), DN3L (CD44 lo CD25 lo ) and DN4 (CD44 lo CD25 -) subsets (Godfrey et al, 1993;Zeng et al, 2007). During the DN1 and DN2 stages, cells receive signals that induce commitment to the T-cell lineage and begin rearrangement of the genomic locus that encodes the T-cell receptor (TCR) b chain.…”

Section: Introductionmentioning

confidence: 99%

Depletion and recovery of lymphoid subsets following morphine administration

Zhang

Xiong

Parker

et al. 2011

British J Pharmacology

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BACKGROUND AND PURPOSEOpioid use and abuse has been linked to significant immunosuppression, which has been attributed, in part, to drug-induced depletion of lymphocytes. We sought to define the mechanisms by which lymphocyte populations are depleted and recover following morphine treatment in mice. EXPERIMENTAL APPROACHMice were implanted with morphine pellets and B-and T-cell subsets in the bone marrow, thymus, spleen and lymph nodes were analysed at various time points. We also examined the effects of morphine on T-cell development using an ex vivo assay. KEY RESULTSThe lymphocyte populations most susceptible to morphine-induced depletion were the precursor cells undergoing selection. As the lymphocytes recovered, more lymphocyte precursors proliferated than in control mice. In addition, peripheral T-cells displayed evidence that they had undergone homeostatic proliferation during the recovery phase of the experiments. CONCLUSIONS AND IMPLICATIONSThe recovery of lymphocytes following morphine-induced depletion occurred in the presence of morphine and via increased proliferation of lymphoid precursors and homeostatic proliferation of T-cells.

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Gads−/− Mice Reveal Functionally Distinct Subsets of TCRβ+ CD4−CD8− Double-Negative Thymocytes

Cited by 14 publications

References 37 publications

In vivo disruption of T cell development by expression of a dominant‐negative polypeptide designed to abolish the SLP‐76/Gads interaction

In vivo disruption of T cell development by expression of a dominant‐negative polypeptide designed to abolish the SLP‐76/Gads interaction

Histone Deacetylase 3 Is Required for Efficient T Cell Development

Depletion and recovery of lymphoid subsets following morphine administration

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