Adhesion- and Degranulation-Promoting Adapter Protein Is Required for Efficient Thymocyte Development and Selection

Wu, Jiansheng; Gheith, Shereen M F; Bezman, Natalie; Liu, Qing Hua; Fostel, Lindsey V.; Swanson, Andrew M.; Freedman, Bruce D.; Koretzky, Gary A.; Peterson, Erik

doi:10.4049/jimmunol.176.11.6681

Cited by 27 publications

(47 citation statements)

References 43 publications

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“…Conjugates are thought to be partially dependent upon LFA-1 for the formation of stable conjugates [50,51]. Thus, our result may reflect the limited effect of GBF on LFA-1 activation, perhaps pointing to SLP-76/Gads-independent pathways to LFA-1 activation and/or the contribution of other factors required for T cell/B cell conjugate formation.…”

Section: Discussionmentioning

confidence: 78%

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: 78%

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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“…Adaptor proteins possess binding sites and domains needed for complex-complex formation (31,32,41). Immune cell-specific adaptors include LAT (linker for activation of T cells), GADS (Grb-2-like adaptor downstream of Shc), SLP-76 (SH2 domain-containing leukocyte protein of 76 kDa), ADAP (adhesion-and degranulation-promoting adaptor protein; previously known as FYN T-binding protein/SLP-76-associated protein [FYB/SLAP]), and SKAP-55 (Src kinase-associated phosphoprotein of 55 kDa; also known as SCAP1) (3,23,24,26,31,32,(40)(41)(42). Phosphorylation of LAT recruits phospholipase C␥1, Grb-2, and GADS-SLP-76 and induces Ca 2ϩ mobilization and cytokine transcription (32,33,41,44).…”

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confidence: 99%

Functional Defects of SKAP-55-Deficient T Cells Identify a Regulatory Role for the Adaptor in LFA-1 Adhesion

Wang

Liu

et al. 2007

Molecular and Cellular Biology

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T-cell receptors (TCRs) undergo microclustering and supramolecular activation cluster formation at the immunological synapse (IS) during conjugation between T cells and antigen-presenting cells (APCs) (5,21,35). Microclustering can in turn activate GTP-binding proteins, protein kinases, phosphatases, and the phosphorylation of adaptor proteins. CD4-and CD8-p56lck activation leads to immunoreceptor tyrosinebased activation motif phosphorylation on the CD3 and TCR chains, the recruitment of ZAP-70 (zeta-associated proteintyrosine kinase of 70 kDa), and the activation of TEC kinases ITK/RLK (interleukin-2-inducible/resting lymphocyte kinase) (1,(31)(32)(33)41). Adaptor proteins possess binding sites and domains needed for complex-complex formation (31,32,41). Immune cell-specific adaptors include LAT (linker for activation of T cells), GADS (Grb-2-like adaptor downstream of Shc), SLP-76 (SH2 domain-containing leukocyte protein of 76 kDa), ADAP (adhesion-and degranulation-promoting adaptor protein; previously known as FYN T-binding protein/SLP-76-associated protein [FYB/SLAP]), and SKAP-55 (Src kinase-associated phosphoprotein of 55 kDa; also known as SCAP1) (3,23,24,26,31,32,(40)(41)(42). Phosphorylation of LAT recruits phospholipase C␥1, Grb-2, and GADS-SLP-76 and induces Ca 2ϩ mobilization and cytokine transcription (32,33,41,44).Binding of leukocyte function-associated antigen 1 (LFA-1; also known as CD11/CD18 or ␣ L ␤ 2 ) to intercellular adhesion molecules 1 and 2 (ICAM-1 and -2) on APCs mediates T-cell-APC conjugation (5,8,11,21,37). Following initial adhesion, TCR/CD3 ligation induces signals (i.e., "inside-out signaling") that further activate integrin adhesion (2,5,8,11,37). Conversion of LFA-1 to intermediate-or higher-affinity forms involves changes in conformation and receptor clustering (11,37). Multiple signaling proteins mediate this process. They include the GTP-binding protein Rap-1, its ligand RapL (regulator of cell adhesion and polarization enriched in lymphoid tissues), RIAM (Rap1-GTP-interacting adaptor molecule), the guanine nucleotide exchange factor Vav-1, and the adaptors ADAP,10,13,15,16,17,18,20,22,27,28,34). The protein-tyrosine kinase ZAP-70 phosphorylates YESP sites in SLP-76, which allows binding to the Src homology 2 (SH2) domain of Vav-1 (29), while the SH2 domain of SLP-76 binds to two YDDV sites in ADAP (30, 39). T-cell lines lacking SLP-76 show impaired superantigen-induced conjugation (44). ADAP is an immune cell-specific adaptor with a unique N-terminal region, a proline-rich region, a canonical and a noncanonical SH3 domain, one Ena/VASP homology 1 (EVH1) binding domain, and two putative nuclear localization motifs (3,4,19,26,31). ADAP is preferentially phosphorylated by the Src kinase p59fynT (4,25) and can cooperate with p59 fynT and SLP-76 in amplifying TCR-induced interleukin-2 (IL-2) transcription (30). The adaptor can up-regulate integrin-mediated adhesion in certain basophilic cell lines (9), while ADAP Ϫ/Ϫ T cells show profound defects in ␤1 and ␤2 integrin clusterin...

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“…1D) [47][48][49][50], we analyzed ADAP −/− and littermate control mice. Similar to slp-76 ace/ace mice, iNKT cells in the spleens and livers of ADAP −/− mice were reduced (Fig.…”

Section: Slpmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…The C-terminal SH2 domain of slp-76 binds to the hematopoietic progenitor kinase HPK-1 [43] and to the adhesion and degranulation-promoting adaptor protein ADAP [44,45]. While HPK-1 inhibits TCR signals [46], ADAP is required for thymocyte selection and TCR-mediated integrinactivation/-expression [47][48][49][50].Using slp-76 ace/ace mice that exhibit a threonine to isoleucine amino acid change at position 428 within the SH2 domain of slp-76 [51, 52], we characterize slp-76 as critical for the regulation of PLZF-and NP-1-expression, cytokine release and the tissue distribution of iNKT cells. While the accumulation of iNKT cells in the thymus and the peripheral lymph nodes is ADAPindependent, interactions of slp-76 with ADAP disrupt the distribution of iNKT cells to the spleens and livers.…”

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confidence: 99%

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A mutation within the SH2 domain of slp‐76 regulates the tissue distribution and cytokine production of iNKT cells in mice

et al. 2016

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TCR ligation is critical for the selection, activation, and integrin expression of T lymphocytes. Here, we explored the role of the TCR adaptor protein slp-76 on iNKT-cell biology. Compared to B6 controls, slp-76 ace/ace mice carrying a missense mutation (Thr428Ile) within the SH2-domain of slp-76 showed an increase in iNKT cells in the thymus and lymph nodes, but a decrease in iNKT cells in spleens and livers, along with reduced ADAP expression and cytokine response. A comparable reduction in iNKT cells was observed in the livers and spleens of ADAP-deficient mice. Like ADAP −/− iNKT cells, slp-76 ace/ace iNKT cells were characterized by enhanced CD11b expression, correlating with an impaired induction of the TCR immediate-early gene Nur77 and a decreased adhesion to ICAM-1. Furthermore, CD11b-intrinsic effects inhibited cytokine release, concanavalin A-mediated inflammation, and iNKT-cell accumulation in the liver. Unlike B6 and ADAP −/− mice, the expression of the transcription factors Id3 and PLZF was reduced, whereas NP-1-expression was enhanced in slp-76 ace/ace mice. Blockade of NP-1 decreased the recovery of iNKT cells from peripheral lymph nodes, identifying NP-1 as an iNKT-cell-specific adhesion factor. Thus, slp-76 contributes to the regulation of the tissue distribution, PLZF, and cytokine expression of iNKT cells via ADAP-dependent and -independent mechanisms.Keywords: ADAP r Cytokine r iNKT cell r Integrin r slp-76Additional supporting information may be found in the online version of this article at the publisher's web-site Eur. J. Immunol. 2016Immunol. . 46: 2121Immunol. -2136 Introduction iNKT cells express a panoply of NK-cell receptors [1] and a canonical TCR through which they recognize (glyco-)lipid antigens [2]. iNKT cells activate similar signaling cascades after TCR ligation like other T lymphocytes [3], but utilize unique transcription factors for their development such as the promyelocytic leukemia zinc finger (PLZF) [4][5][6]. According to two different developmental models PLZF characterizes distinct maturation stages and polarized subsets. The sequential lineage model suggests a gradual decrease of PLZFexpression following selection of iNKT cells which show a Th2-dominated cytokine profile during earlier and a Th1-dominated cytokine profile during later stages of intrathymic maturation [1,7]. The second model describes lineage diversification and simultaneous differentiation into Th1-, Th2-, or Th17-polarized subsets that are defined by the level of PLZF-expression [8]. Although many iNKT cells release both Th1 and Th2 cytokines on a single cell level [9], the production of IL-17 and IFN-γ is mutually exclusive within NK1.1 − cells [10][11][12]. Several transcription factors, such as Egr2, T-bet, ThPOK, Id2, Id3, and the Tec kinases Itk and Rlk have been implicated in the differentiation of iNKT cell subsets [6,8,[13][14][15][16][17] which home to distinct tissues. Specifically, liver and spleen constitute the main source for the Th1-polarized sublineage which is PLZF low . Th2-or ...

show abstract

Adhesion- and Degranulation-Promoting Adapter Protein Is Required for Efficient Thymocyte Development and Selection

Cited by 27 publications

References 43 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

Functional Defects of SKAP-55-Deficient T Cells Identify a Regulatory Role for the Adaptor in LFA-1 Adhesion

A mutation within the SH2 domain of slp‐76 regulates the tissue distribution and cytokine production of iNKT cells in mice

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