Detection and Characterizationof Cellular Immune Responses Using Peptide–MHC Microarrays
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{"title"=>"Detection and characterization of cellular immune responses using peptide-MHC microarrays", "type"=>"journal", "authors"=>[{"first_name"=>"Yoav", "last_name"=>"Soen", "scopus_author_id"=>"6602347180"}, {"first_name"=>"Daniel S.", "last_name"=>"Chen", "scopus_author_id"=>"16165697600"}, {"first_name"=>"Daniel L.", "last_name"=>"Kraft", "scopus_author_id"=>"35748684200"}, {"first_name"=>"Mark M.", "last_name"=>"Davis", "scopus_author_id"=>"7404850298"}, {"first_name"=>"Patrick O.", "last_name"=>"Brown", "scopus_author_id"=>"35414776900"}], "year"=>2003, "source"=>"PLoS Biology", "identifiers"=>{"scopus"=>"2-s2.0-2942730105", "sgr"=>"2942730105", "issn"=>"15449173", "isbn"=>"1545-7885 (Electronic)\r1544-9173 (Linking)", "pmid"=>"14691537", "doi"=>"10.1371/journal.pbio.0000065", "pui"=>"39112141"}, "id"=>"2e578a18-f634-370e-b353-b1a6684956c0", "abstract"=>"The detection and characterization of antigen-specific T cell populations is critical for understanding the development and physiology of the immune system and its responses in health and disease. We have developed and tested a method that uses arrays of peptide-MHC complexes for the rapid identification, isolation, activation, and characterization of multiple antigen-specific populations of T cells. CD4(+) or CD8(+) lymphocytes can be captured in accordance with their ligand specificity using an array of peptide-MHC complexes printed on a film-coated glass surface. We have characterized the specificity and sensitivity of a peptide-MHC array using labeled lymphocytes from T cell receptor transgenic mice. In addition, we were able to use the array to detect a rare population of antigen-specific T cells following vaccination of a normal mouse. This approach should be useful for epitope discovery, as well as for characterization and analysis of multiple epitope-specific T cell populations during immune responses associated with viral and bacterial infection, cancer, autoimmunity, and vaccination.", "link"=>"http://www.mendeley.com/research/detection-characterization-cellular-immune-responses-using-peptidemhc-microarrays", "reader_count"=>6, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>1, "Student > Bachelor"=>1, "Unspecified"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>1, "Student > Bachelor"=>1, "Unspecified"=>1}, "reader_count_by_subject_area"=>{"Materials Science"=>1, "Medicine and Dentistry"=>1, "Agricultural and Biological Sciences"=>2, "Immunology and Microbiology"=>1, "Unspecified"=>1}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>1, "United Kingdom"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/964649"], "description"=>"<div><p>(A) A schematic diagram illustrating T cell binding to its cognate peptide–MHC spot via peptide–MHC–TCR interactions (not drawn to scale).</p>\n <p>(B) Cells immobilized on the cellular microarray are immediately visible to inspection. When cell coverage exceeds approximately 20% confluency, the cluster becomes visible to the naked eye. Shown are cell clusters bound to several triplicate spots of either peptide–MHC complexes or antibodies. The image was taken using a digital camera. The arrows and legends indicate the locations and identities of each of the printed triplicates. Spot diameter and interspot distance are about 400 μm and 700 μm, respectively. Scale bar is 1.4 mm in length.</p></div>", "links"=>[], "tags"=>["cellular"], "article_id"=>634576, "categories"=>["Biotechnology", "Microbiology", "Infectious Diseases", "Immunology", "Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0000065.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_Cellular_Microarray_/634576", "title"=>"A Cellular Microarray", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:20:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/964719"], "description"=>"<div><p>(A) DiD-labeled OT-1 OVA-specific cells (red) were diluted 1:100 (top panels) or 1:1,000 (bottom panels) in DiO-labeled, monocyte-depleted C57BL/6 mouse lymph node cells (green). DiD-labeled OT-1 cells of 2.7 × 10<sup>4</sup> (top panels) and 4.41 × 10<sup>3</sup> (bottom panels) were mixed, respectively, with 2.7 × 10<sup>6</sup> and 4.41 × 10<sup>6</sup> of DiO-labeled CD11b-depleted lymph node cells. The different dilutions were applied to identical but separate arrays printed with the OVA/K<sup>b</sup> tetramer, the LCMV/K<sup>d</sup> control tetramer, and three different antibody spots (mouse anti-CD8, mouse anti-CD4, and mouse anti-CD28). Following 10-min incubation at 37°C, the cells were washed in RPMI and the slide was scanned. Shown are the OT-1 (red) and lymph node (green) cells that were captured by the OVA tetramer (left), LCMV tetramer (second left), and the antibody spots.</p>\n <p>(B) Class II MHC-mediated detection and sorting of helper T cells. DiD-labeled 5c.c7 lymphocytes (3.5 × 10<sup>4</sup>) (red) were diluted 100-fold in 3.6 × 10<sup>6</sup> B10A DiO splenocytes (green). The mixed suspension was incubated at 37°C for 10 min prior to wash with RPMI. Shown are the 5c.c7 sells (red) and splenocytes (green) captured by the MCC (left) and anti-CD28 (right) spots.</p></div>", "links"=>[], "tags"=>["tetramer-mediated", "detection", "ova-specific", "ctls", "mcc-specific", "helper"], "article_id"=>634642, "categories"=>["Biotechnology", "Microbiology", "Infectious Diseases", "Immunology", "Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0000065.g003", "stats"=>{"downloads"=>1, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sensitivity_of_Peptide_8211_MHC_Tetramer_Mediated_Detection_of_OVA_Specific_CTLs_and_MCC_Specific_Helper_T_Cells_/634642", "title"=>"Sensitivity of Peptide–MHC Tetramer-Mediated Detection of OVA-Specific CTLs and MCC-Specific Helper T Cells", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:21:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/474064", "https://ndownloader.figshare.com/files/474069", "https://ndownloader.figshare.com/files/474077", "https://ndownloader.figshare.com/files/474085"], "description"=>"<div><p>The detection and characterization of antigen-specific T cell populations is critical for understanding the development and physiology of the immune system and its responses in health and disease. We have developed and tested a method that uses arrays of peptide–MHC complexes for the rapid identification, isolation, activation, and characterization of multiple antigen-specific populations of T cells. CD4<sup>+</sup> or CD8<sup>+</sup> lymphocytes can be captured in accordance with their ligand specificity using an array of peptide–MHC complexes printed on a film-coated glass surface. We have characterized the specificity and sensitivity of a peptide–MHC array using labeled lymphocytes from T cell receptor transgenic mice. In addition, we were able to use the array to detect a rare population of antigen-specific T cells following vaccination of a normal mouse. This approach should be useful for epitope discovery, as well as for characterization and analysis of multiple epitope-specific T cell populations during immune responses associated with viral and bacterial infection, cancer, autoimmunity, and vaccination.</p> </div>", "links"=>[], "tags"=>["detection", "characterizationof", "cellular", "responses", "microarrays"], "article_id"=>153397, "categories"=>["Biotechnology", "Cancer", "Microbiology", "Immunology", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>["https://dx.doi.org/10.1371/journal.pbio.0000065.sg003", "https://dx.doi.org/10.1371/journal.pbio.0000065.sg001", "https://dx.doi.org/10.1371/journal.pbio.0000065.sg002", "https://dx.doi.org/10.1371/journal.pbio.0000065.sd001"], "stats"=>{"downloads"=>16, "page_views"=>106, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Detection_and_Characterizationof_Cellular_Immune_Responses_Using_Peptide_MHC_Microarrays/153397", "title"=>"Detection and Characterizationof Cellular Immune Responses Using Peptide–MHC Microarrays", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-01-20 11:44:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/964757"], "description"=>"<div><p>CTLs from OVA-vaccinated and mock-vaccinated mice were analyzed in parallel using an MHC array and flow cytometry analysis. Mice received base-of-tail injections of OVA/Freund's adjuvant emulsions or PBS (mock)/Freund's adjuvant on day 0 and day 7. Draining lymph nodes were harvested on day 11, and dissociated into a single-cell suspension.</p>\n <p>(A) CTLs were enriched on an anti-CD8-bead column. Cells (2 × 10<sup>6</sup> and 3.2 × 10<sup>6</sup>) from the OVA- and mock-vaccinated mice, respectively, were incubated on identical but separate arrays printed with OVA/K<sup>b</sup> tetramer, LCMV/K<sup>d</sup> (control) tetramer, and various antibodies. The three panels show the relevant array results. A rare population of cells from the OVA-vaccinated mouse was captured on the OVA/K<sup>b</sup> (left), but not on the LCMV/K<sup>d</sup> (right), tetramer spot. The cells captured on the OVA/K<sup>b</sup> tetramer spot were CD8<sup>+</sup> (as determined by counter-staining using an anti-CD8–FITC mAb on the array). An arrowhead points to some of the cells that were bound to that spot. The cells from the mock-vaccinated mouse did not bind the OVA tetramer (middle spot) or the LCMV/K<sup>d</sup> tetramer (data not shown). Spot regions are marked with a blue color by overlaying the tetramer's PE fluorescent signal onto the DIC image.</p>\n <p>(B) FACS analysis of the cells from the OVA-vaccinated (left panel) and mock-vaccinated mice (right). Of the total CD8<sup>+</sup> cells from the vaccinated mouse, 0.27% co-stain with OVA/K<sup>b</sup>, compared with 0.01% in the mock-vaccinated mouse.</p></div>", "links"=>[], "tags"=>["biotechnology", "oncology", "cell biology", "immunology", "Infectious diseases", "microbiology", "molecular biology"], "article_id"=>634680, "categories"=>["Biotechnology", "Microbiology", "Infectious Diseases", "Immunology", "Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0000065.g004", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_a_Weak_Immune_Response_to_Vaccination_/634680", "title"=>"Detection of a Weak Immune Response to Vaccination", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:21:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/964690"], "description"=>"<p>Prelabeled, OVA-specific OT-1 (green) and MCC-specific 5c.c7 (red) lymphocytes (6 ×10<sup>5</sup>) were mixed at a 1:1 ratio and added to the preprinted peptide–MHC array. While anti-CD3 and anti-CD28 mAb spots (left and right top panels) immobilize both lymphocyte populations, the OVA and MCC tetramers (left and right bottom panels) capture only the OVA-specific and MCC-specific cells, respectively. Cells were labeled with DiO and DiD lipophilic tracers that incorporate into the cell membrane and do not affect the specificity of cell capture. The printed peptide–MHC tetramer spots contain the fluorescent dye, which allows visualization of peptide–MHC spot borders (data not shown).</p>", "links"=>[], "tags"=>["lymphocyte", "immobilization"], "article_id"=>634617, "categories"=>["Biotechnology", "Microbiology", "Infectious Diseases", "Immunology", "Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0000065.g002", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Specificity_of_Lymphocyte_Immobilization_by_Peptide_8211_MHC_Arrays_/634617", "title"=>"Specificity of Lymphocyte Immobilization by Peptide–MHC Arrays", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:20:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/964792"], "description"=>"<div><p>OT-1 lymphocytes were preloaded with Fura-2 dye for 20 min at room temperature prior to exposure to the cellular microarray (time 0). The cells were incubated at 37°C, 5% CO<sub>2</sub>, and fluorescence intensities at 340 nm and 380 nm were measured every 30 s from each of the spots on the array. Calcium flux signal was triggered in the OVA-specific cells as soon as they reached the OVA/K<sup>b</sup> tetramer spots, the activating antibody spots (anti-CD3 and anti-CD3/anti-CD28), but not the irrelevant MCC/E<sup>k</sup> tetramer or the nonactivating antibody spots (e.g. anti-CD28).</p>\n <p>(A) Calcium flux signal (yellow) overlaid onto a 10x DIC image of OT-1 cells on a combined FITC-labeled anti-CD3/anti-CD28 spot. Fura-2 fluorescence intensity ratios are represented in pseudo-color, and the overlaid spot region (blue) is determined from the FITC image. Note that the spot pattern can be inferred from the Fura-2 signal representing activation, even without removing unbound cells.</p>\n <p>(B) A time-lapsed series of images taken of a representative OT-1 lymphocyte specifically immobilized onto an OVA/K<sup>b</sup> spot. The cell undergoes a rapid, transient calcium flux as measured by Fura-2 fluorescence.</p>\n <p>(C) Averaged calcium flux traces (<i>n</i> = 3) recorded from an OVA/K<sup>b</sup> spot (purple circles), anti-CD3/anti-CD28 spot (brown triangles), anti-CD3 spot (green circles), and nonactivating anti-CD28 spot (orange triangles). Error bars represent calculated standard error of mean.</p></div>", "links"=>[], "tags"=>["ot-1", "lymphocytes", "preprinted", "spots", "mhc"], "article_id"=>634719, "categories"=>["Biotechnology", "Microbiology", "Infectious Diseases", "Immunology", "Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Yoav Soen", "Daniel S Chen", "Daniel L Kraft", "Mark M Davis", "Patrick O Brown"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0000065.g005", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Activation_of_OT_1_Lymphocytes_Following_an_Exposure_to_Preprinted_Spots_on_an_MHC_Array_/634719", "title"=>"Activation of OT-1 Lymphocytes Following an Exposure to Preprinted Spots on an MHC Array", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:22:29"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"3", "full-text"=>"2", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2016", "month"=>"9"}
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  • {"unique-ip"=>"9", "full-text"=>"8", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"6", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"4", "full-text"=>"3", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"7", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"7", "full-text"=>"6", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
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Relative Metric

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