The Ras/MAPK Pathway Is Required for Generation of iNKT Cells
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{"title"=>"The Ras/MAPK pathway is required for generation of iNKT cells", "type"=>"journal", "authors"=>[{"first_name"=>"Taishan", "last_name"=>"Hu", "scopus_author_id"=>"55434250500"}, {"first_name"=>"Idoia", "last_name"=>"Gimferrer", "scopus_author_id"=>"6603194908"}, {"first_name"=>"Amie", "last_name"=>"Simmons", "scopus_author_id"=>"35488997200"}, {"first_name"=>"David", "last_name"=>"Wiest", "scopus_author_id"=>"7005472565"}, {"first_name"=>"José", "last_name"=>"Alberola-Ila", "scopus_author_id"=>"7004285673"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"21572967", "sgr"=>"79955910259", "doi"=>"10.1371/journal.pone.0019890", "scopus"=>"2-s2.0-79955910259", "pui"=>"361753367", "issn"=>"19326203"}, "id"=>"e3235de0-a51b-30b8-af4c-33a70c3c9a62", "abstract"=>"iNKT cells derive from CD4(+)CD8(+) DP thymocytes, and are selected by thymocyte-thymocyte interactions through signals from their invariant Vα14-Jα18 TCR and from the costimulatory molecules SLAMF1 and SLAMF6. Genetic studies have demonstrated the contribution of different signaling pathways to this process. Surprisingly, current models imply that the Ras/MAPK pathway, one of the critical mediators of conventional αβ T cell positive selection, is not necessary for iNKT cell development. Using mice defective at different levels of this pathway our results refute this paradigm, and demonstrate that Ras, and its downstream effectors Egr-1 and Egr-2 are required for positive selection of iNKT cells. Interestingly our results also show that there are differences in the contributions of several of these molecules to the development of iNKT and conventional αβ T cells.", "link"=>"http://www.mendeley.com/research/rasmapk-pathway-required-generation-inkt-cells", "reader_count"=>12, "reader_count_by_academic_status"=>{"Researcher"=>4, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Other"=>1}, "reader_count_by_user_role"=>{"Researcher"=>4, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Other"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>5, "Medicine and Dentistry"=>1, "Arts and Humanities"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United Kingdom"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/777269"], "description"=>"<p>(<b>a</b>) Thymic profile of WT and dnRas mice (<b>b</b>) Percentages and absolute numbers of iNKT cells in the thymus (T), spleen (S) and liver mononuclear cells (L) of normal littermate controls (WT) and dnRas mice stained with CD4, CD8, PBS57-loaded CD1d tetramer and TCRβ. (<b>c</b>) Percentages and absolute numbers of iNKT cell subpopulations in gated TCR<sup>hi</sup>PBS57-CD1dtet<sup>+</sup> thymocytes from WT and dnRas mice. Results representative of nine independent dnRas and WT pairs analyzed in five experiments<b>,</b> except for the CD44/NK1.1 histograms (n = 2). The bar graphs show the average and SEM of all the experiments. Significance as assessed using a two-tailed unpaired t-test. ***<0.001, **<0.01. (<b>d</b>) Gating strategy used to sort the different populations. (<b>e</b>) Expression of Egr-1, Egr-2 and Id3 in sorted Tet<sup>+</sup> HSA<sup>hi</sup> and Tet<sup>+</sup> HSA<sup>lo</sup> from normal littermate control (WT) and dnRas mice. Bar graphs show relative expression of dnRas compared to WT for three independent experiments. Each experiment was an independent sort of a WT and dnRas pair. Expression in each experiment was normalized to the expression levelis in Tet<sup>+</sup> HSA<sup>hi</sup> WT cells. Significance as assessed using a two-tailed unpaired t-test. **<0.01.</p>", "links"=>[], "tags"=>["mice", "inkt"], "article_id"=>447636, "categories"=>["Molecular Biology", "Developmental Biology", "Immunology"], "users"=>["Taishan Hu", "Idoia Gimferrer", "Amie Simmons", "David Wiest", "José Alberola-Ila"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0019890.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_dnRas_mice_lack_iNKT_cells_/447636", "title"=>"dnRas mice lack iNKT cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 18:37:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/777354"], "description"=>"<p>(<b>a</b>) Contribution of WT and Egr1<sup>-/-</sup> (Egr1KO) cells to the iNKT compartment in thymus (T), spleen (S) and liver (L) of mixed bone marrow chimeras generated by injecting Egr1<sup>-/-</sup> (CD45.2) and F1(C57BL/6xB6-LY5.2/Cr) (CD45.1;CD45.2) bone marrow cells into lethally irradiated B6-LY5.2/Cr recipient mice (CD45.1). Mean and SEM is shown on the right (n = 5). Significance was assessed using a paired t-test. **<0.01, *<0.05. This is one out of two independent experiments. <b>(b)</b> Percentages and absolute numbers of iNKT cells in the thymus (T), spleen (S) and liver mononuclear cells (L) of WT and Egr-2<sup>f/f</sup>-<i>lck</i>-Cre (EGR2KO) mice stained with CD4, CD8, PBS57-loaded CD1d tetramer and TCRβ. <b>(c)</b> Percentages and absolute numbers of iNKT cell subpopulations in gated TCR-β<sup>hi</sup>PBS57-CD1dtet<sup>+</sup> thymocytes from WT and EGR2KO mice. Results representative of five independent pairs in three independent experiments. The bar graphs show the average and SEM of all the experiments. Significance was assessed using an unpaired t-test. ***<0.001, **<0.01.</p>", "links"=>[], "tags"=>["egr2", "quantitatively", "inkt"], "article_id"=>447725, "categories"=>["Molecular Biology", "Developmental Biology", "Immunology"], "users"=>["Taishan Hu", "Idoia Gimferrer", "Amie Simmons", "David Wiest", "José Alberola-Ila"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0019890.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Egr1_and_Egr2_contribute_in_a_quantitatively_different_manner_to_iNKT_cell_development_/447725", "title"=>"Egr1 and Egr2 contribute in a quantitatively different manner to iNKT cell development.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 18:37:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/777460"], "description"=>"<p><b>(a)</b> Donor contributions in bone marrow chimeras generated by injecting MiG-Bcl2 transduced Egr2<sup>f/f</sup><i>lck</i>-Cre (Egr2KO) (CD45.2) and wild-type (CD45.1;CD45.2) cells into lethally irradiated CD45.1 recipients, analyzed at 8–12 weeks (left), and a representative histogram of GFP expression in MiG-Bcl2 transduced Egr2KO thymocytes (right). <b>(b)</b> TcR thymic profiles (upper panel) and iNKT cells frequency (lower panels) in wild-type, Egr2KO and Egr2KO-Bcl2 donor cells in the thymus (T) and Liver (L) of chimeras described in (a). <b>(c)</b> Percentages of different cell subpopulations from WT Egr2KO and Egr2KO;Bcl-2 in the mixed bone marrow chimeras mice. Results representative of eight independent chimeras in two independent experiments. The bar graphs show the average and SEM of all the experiments. Significance as assessed using the unpaired t-test. ***<0.001.</p>", "links"=>[], "tags"=>["inkt", "defect"], "article_id"=>447831, "categories"=>["Molecular Biology", "Developmental Biology", "Immunology"], "users"=>["Taishan Hu", "Idoia Gimferrer", "Amie Simmons", "David Wiest", "José Alberola-Ila"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0019890.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bcl_2_can_not_rescue_the_iNKT_defect_in_Egr2KOmice_/447831", "title"=>"Bcl-2 can not rescue the iNKT defect in Egr2KOmice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 18:38:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/777590"], "description"=>"<p>(<b>a</b>) Thymic profile of littermate wild-type (WT), or Egr1<sup>-/-</sup>;Egr2<sup>f/f</sup><i>lck</i>cre (DKO) mice <b>(b)</b> Percentages and absolute numbers of iNKT cells in the thymus (T), spleen (S) and liver mononuclear cells (L) of WT and EgrDKO mice stained with CD4, CD8, PBS57-loaded CD1d tetramer and TCRβ. <b>(c)</b> Percentages and absolute numbers of iNKT cell subpopulations in gated thymic TCR-βhiPBS57-CD1dtet<sup>+</sup> of WT and EgrDKO mice. Results representative of five independent pairs in three independent experiments. The bar graphs show the average and SEM of all the experiments. Significance as assessed using the unpaired t-test. ***<0.001, **<0.01</p>", "links"=>[], "tags"=>["mice", "inkt"], "article_id"=>447958, "categories"=>["Molecular Biology", "Developmental Biology", "Immunology"], "users"=>["Taishan Hu", "Idoia Gimferrer", "Amie Simmons", "David Wiest", "José Alberola-Ila"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0019890.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Egr1_Egr2_f_f_lck_cre_mice_have_a_complete_block_in_iNKT_development_/447958", "title"=>"Egr1<sup>-/-</sup>; Egr2<sup>f/f</sup><i>lck</i>cre mice have a complete block in iNKT development.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 18:38:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/777728"], "description"=>"<p>Slamf1, Slamf3, Slamf6, Slamf5 and CD1d expression levels in DP thymocytes mice were assessed by flow cytometry. Shown are representative histograms, and the mean and SEM of the normalized MFI of DP populations. In <b>(A)</b> WT vs. dnRas (n = 5). In <b>(B)</b> WT vs. Egr1<sup>-/-</sup>;Egr2<sup>f/f</sup>-<i>lck</i>-Cre (Egr-DKO) (n = 3). <b>(C)</b> SAP and Bcl<sub>xL</sub> expression levels in DP thymocytes from WT or dnRas mice were assessed by intracellular flow cytometry. Shown are representative histograms, and the mean and SEM of the normalized MFI of DP populations (n = 5). To normalize the MFI, we averaged the MFI for the WT mice in each experiment and considered that value 1. The bar graphs show the average and SEM of all the experiments. Significance was assessed using the unpaired t-test ***<0.001, **<0.01 *<0.05<b>.</b></p>", "links"=>[], "tags"=>["slamf6", "cd1d", "knockout"], "article_id"=>448095, "categories"=>["Molecular Biology", "Developmental Biology", "Immunology"], "users"=>["Taishan Hu", "Idoia Gimferrer", "Amie Simmons", "David Wiest", "José Alberola-Ila"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0019890.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Defects_in_Slamf1_Slamf6_and_CD1d_expression_in_dnRas_but_not_Egr_1_2_double_knockout_mice_/448095", "title"=>"Defects in Slamf1, Slamf6 and CD1d expression in dnRas, but not Egr-1,2 double knockout mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 18:39:32"}

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Relative Metric

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