Low 2-Dimensional CD4 T Cell Receptor Affinity for Myelin Sets in Motion Delayed Response Kinetics
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{"title"=>"Low 2-dimensional CD4 T cell receptor affinity for myelin sets in motion delayed response kinetics", "type"=>"journal", "authors"=>[{"first_name"=>"Kristen M.", "last_name"=>"Rosenthal", "scopus_author_id"=>"26032351200"}, {"first_name"=>"Lindsay J.", "last_name"=>"Edwards", "scopus_author_id"=>"23484826200"}, {"first_name"=>"Joseph J.", "last_name"=>"Sabatino", "scopus_author_id"=>"24336484800"}, {"first_name"=>"Jennifer D.", "last_name"=>"Hood", "scopus_author_id"=>"24171053900"}, {"first_name"=>"Heather A.", "last_name"=>"Wasserman", "scopus_author_id"=>"35207535000"}, {"first_name"=>"Cheng", "last_name"=>"Zhu", "scopus_author_id"=>"55584801040"}, {"first_name"=>"Brian D.", "last_name"=>"Evavold", "scopus_author_id"=>"7004114695"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"364373839", "doi"=>"10.1371/journal.pone.0032562", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22412888", "issn"=>"19326203", "scopus"=>"2-s2.0-84857853669", "sgr"=>"84857853669"}, "id"=>"569fc21b-373e-3ae0-af03-affd681be99c", "abstract"=>"T cells recognizing self-peptides that mediate autoimmune disease and those that are responsible for efficacious immunity against pathogens may differ in affinity for antigen due to central and peripheral tolerance mechanisms. Here we utilize prototypical self-reactive (myelin) and viral-specific (LCMV) T cells from T cell receptor (TCR) transgenic mice (2D2 and SMARTA, respectively) to explore affinity differences. The T cells responsive to virus possessed >10,000 fold higher 2D affinity as compared to the self-reactive T cells. Despite their dramatically lower affinity for their cognate ligand, 2D2 T cells respond with complete, albeit delayed, activation (proliferation and cytokine production). SMARTA activation occurs rapidly, achieving peak phosphorylation of p38 (1 minute), Erk (30 minutes), and Jun (3 hours) as well as CD69 and CD25 upregulation (3 and 6 hours, respectively), with a corresponding early initiation of proliferation. 2D2 stimulation with MOG results in altered signaling--no phospho-Erk or phospho-p38 accumulation, significantly delayed activation kinetics of Jun (12 hours), and delayed but sustained SHP-1 activity--as well as delayed CD69 and CD25 expression (12-24 hours), and slow initiation of proliferation. This delay was not intrinsic to the 2D2 T cells, as a more potent antigen with >100-fold increased 2D affinity restored rapid response kinetics in line with those identified for the viral antigen. Taken together, these data demonstrate that time can offset low TCR affinity to attain full activation and suggest a mechanism by which low affinity T cells participate in autoimmune disease.", "link"=>"http://www.mendeley.com/research/low-2dimensional-cd4-t-cell-receptor-affinity-myelin-sets-motion-delayed-response-kinetics", "reader_count"=>17, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Other"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Other"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>3, "Neuroscience"=>2, "Chemistry"=>1, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Neuroscience"=>{"Neuroscience"=>2}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Czech Republic"=>1, "United States"=>2, "Germany"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/671212"], "description"=>"<p>(A) Splenocytes from SMARTA or 2D2 mice were stimulated with 10 µM of the indicated antigen and the percent of maximal Erk phosphorylation was assessed at various time points by flow cytometry (gated on CD4<sup>+</sup> cells). (B) Splenocytes from SMARTA or 2D2 mice were stimulated with 10 µM of the indicated antigen and the percent of maximal p38 MAPK phosphorylation was assessed at various time points by flow cytometry (gated on CD4<sup>+</sup> cells). (C) Splenocytes from SMARTA or 2D2 mice were stimulated with 10 µM of the indicated antigen and SHP-1 phosphatase activity was assessed at various time points using a colorimetric assay for free phosphate with a phosphorylated peptide substrate specific for SHP-1. The percent of maximal response was assessed to highlight the kinetics of the signaling response and was calculated using GraphPad Prism/ All experiments were repeated at least three times.</p>", "links"=>[], "tags"=>["cells", "stimulated", "mog", "altered", "signaling"], "article_id"=>341693, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g004", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2D2_T_cells_stimulated_with_MOG_have_an_altered_signaling_program_/341693", "title"=>"2D2 T cells stimulated with MOG have an altered signaling program.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:02:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/671513"], "description"=>"<p>Splenocytes were harvested and dose response curves were generated with the indicated peptides up to a maximal concentration of 100 µM to determine the proliferative capacity of 2D2 CD4<sup>+</sup> T cells. The nested sets of peptides were generated from the known full length MOG<sub>35–55</sub> epitope. Three pluses represent proliferation similar to the parent epitope, with each deduction of a plus representing a log shift in the dose required for maximal proliferation. A minus represents no proliferation above background.</p>", "links"=>[], "tags"=>["2d2", "mog", "epitope"], "article_id"=>342002, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.t001", "stats"=>{"downloads"=>0, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_2D2_core_MOG_epitope_is_MOG_39_8211_48_/342002", "title"=>"The 2D2 core MOG epitope is MOG<sub>39–48</sub>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 08:04:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/671092"], "description"=>"<p>Splenocytes from SMARTA or 2D2 mice were stimulated with 10 µM of the indicated antigen and signaling events were assessed. (A) A representative plot of pErk expression assessed at various time points by flow cytometry, gated on CD4<sup>+</sup> cells. (B) A representative plot of p-c-Jun expression assessed at various time points by flow cytometry, gated on CD4<sup>+</sup> cells. Graphical representation of averaged p-Erk (C) and p-c-Jun (D) expression are from at least three independent experiments at various time points. (E) Cell proliferation was assessed after treatment with the Erk-specific MEK inhibitor U0126. All experiments were repeated at least three times.</p>", "links"=>[], "tags"=>["cells", "stimulated", "mog", "detectible", "perk", "delayed", "phosphorylation"], "article_id"=>341574, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g003", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2D2_cells_stimulated_with_MOG_have_no_detectible_pErk_and_delayed_phosphorylation_of_c_Jun_/341574", "title"=>"2D2 cells stimulated with MOG have no detectible pErk and delayed phosphorylation of c-Jun.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:02:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/670896"], "description"=>"<p>(A) 2D2 or SMARTA T cells were stained with antigen-specific I-A<sup>b</sup> tetramer (MOG<sub>38–49</sub> or gp<sub>66–77</sub>, respectively) or a negative control I-A<sup>b</sup> tetramer and analyzed by flow cytometry, gated on CD4<sup>+</sup> cells. (B) Human RBCs were coated with the indicated pMHC complex and brought into contact with the corresponding T cell by micropipette numerous times. The resulting adhesion frequency was used to derive the effective 2D affinity (<i>A<sub>c</sub>K<sub>a</sub></i>, in µm<sup>4</sup>). All experiments were performed at least three times.</p>", "links"=>[], "tags"=>["2d", "affinity", "2d2", "smarta", "cells"], "article_id"=>341368, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_effective_2D_affinity_of_2D2_and_SMARTA_CD4_T_cells_differs_/341368", "title"=>"The effective 2D affinity of 2D2 and SMARTA CD4<sup>+</sup> T cells differs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:01:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/671320"], "description"=>"<p>SMARTA or 2D2 splenocytes were stimulated with 10 µM antigen for various time points and CD69 (A) and CD25 (B) expression on CD4<sup>+</sup> cells was analyzed by flow cytometry. SMARTA or 2D2 splenocytes were stimulated with various concentrations of antigen for 24 hrs. and CD69 (C) and CD25 (D) expression on CD4<sup>+</sup> cells was analyzed by flow cytometry. Experiments were performed at least three times.</p>", "links"=>[], "tags"=>["stimulation", "mog", "delayed", "activation"], "article_id"=>341807, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g005", "stats"=>{"downloads"=>1, "page_views"=>64, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2D2_CD4_T_cell_stimulation_with_MOG_results_in_delayed_expression_of_activation_markers_/341807", "title"=>"2D2 CD4<sup>+</sup> T cell stimulation with MOG results in delayed expression of activation markers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:03:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/671436"], "description"=>"<p>Splenocytes from SMARTA or 2D2 mice were CFSE labeled and stimulated with 10 µM of the indicated antigen for various times. (A) Representative plots from at least three independent experiments show CFSE dilution of CD4<sup>+</sup> cells, assessed by flow cytometry, to detect proliferation at indicated times. (B) On various days, total CD4<sup>+</sup> T cell numbers were assessed using BD Trucount tubes (BD Biosciences) to gauge cellular accumulation after peptide stimulation (On Day 7, SMARTA:gp61 and 2D2:NFM, p = 0.1; SMARTA:gp61 and 2D2:MOG p = 0.5; 2D2:NFM and 2D2:MOG p = 0.2). (C) After the indicated APC contact time, CD4+ T cells were MACS purified to remove the cells from the APCs. The percent of CD25+ and CD69+ CD4+ T cells was assessed by flow cytometry at 24 hours, as indicated on the x-axis. Experiments were performed at least three times; p values were generated using student's t-test on GraphPad Prism.</p>", "links"=>[], "tags"=>["stimulation", "delayed", "initiation", "proliferation", "eventual", "accumulation"], "article_id"=>341917, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g006", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MOG_stimulation_results_in_delayed_initiation_of_proliferation_but_eventual_accumulation_of_CD4_T_cells_/341917", "title"=>"MOG stimulation results in delayed initiation of proliferation but eventual accumulation of CD4<sup>+</sup> T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:04:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/670997"], "description"=>"<p>(A) 6×10<sup>5</sup> splenocytes from SMARTA or 2D2 mice were stimulated with various doses of the indicated antigen for 72 hrs. and <sup>3</sup>H-Thy was added during the last 18 hrs. to assess proliferation. (B) The concentration of peptide needed to reach the half-maximal response (EC<sub>50</sub> values, µM) were derived from the above proliferation assay using GraphPad Prism. (C) The reciprocal EC<sub>50</sub> was plotted against the effective 2D affinity. The open circle for 2D2:MOG denotes the uncertainty of 2D2:MOG affinity, as it was below 10<sup>−8</sup> µm<sup>4</sup>, the limit of detection for this assay. (D) 1.5×10<sup>6</sup> splenocytes were stimulated for 24 hrs. and supernatants were harvested to determine the amount of IL-2 by ELISA. (E) 1.5×10<sup>6</sup> splenocytes were stimulated for 24 hrs. and supernatants were harvested to determine the amount of IFN- γ by ELISA. (F) 1.5×10<sup>6</sup> splenocytes were stimulated with 10 µM of the indicated antigen and supernatants were harvested (24 hrs. for IL-2 and IFN-γ or 48 hrs. for IL-4) to determine the amount of cytokine by ELISA (<i>P</i> value: * = 0.025, ** = 0.005, ***<0.002). All experiments were performed at least three times.</p>", "links"=>[], "tags"=>["splenocytes", "stimulated", "mog"], "article_id"=>341487, "categories"=>["Molecular Biology", "Immunology"], "users"=>["Kristen M. Rosenthal", "Lindsay J. Edwards", "Joseph J. Sabatino Jr", "Jennifer D. Hood", "Heather A. Wasserman", "Cheng Zhu", "Brian D. Evavold"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0032562.g002", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2D2_splenocytes_stimulated_with_MOG_have_a_low_functional_avidity_/341487", "title"=>"2D2 splenocytes stimulated with MOG have a low functional avidity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:01:44"}

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Relative Metric

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