Overnight Resting of PBMC Changes Functional Signatures of Antigen Specific T- Cell Responses: Impact for Immune Monitoring within Clinical Trials
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{"title"=>"Overnight Resting of PBMC Changes Functional Signatures of Antigen Specific T- Cell Responses: Impact for Immune Monitoring within Clinical Trials", "type"=>"journal", "authors"=>[{"first_name"=>"Sarah", "last_name"=>"Kutscher", "scopus_author_id"=>"25621301100"}, {"first_name"=>"Claudia J.", "last_name"=>"Dembek", "scopus_author_id"=>"25621368600"}, {"first_name"=>"Simone", "last_name"=>"Deckert", "scopus_author_id"=>"55879880300"}, {"first_name"=>"Carolina", "last_name"=>"Russo", "scopus_author_id"=>"55309177500"}, {"first_name"=>"Nina", "last_name"=>"Körber", "scopus_author_id"=>"55880042800"}, {"first_name"=>"Johannes R.", "last_name"=>"Bogner", "scopus_author_id"=>"7005719945"}, {"first_name"=>"Fabian", "last_name"=>"Geisler", "scopus_author_id"=>"7005793867"}, {"first_name"=>"Andreas", "last_name"=>"Umgelter", "scopus_author_id"=>"23026255800"}, {"first_name"=>"Michael", "last_name"=>"Neuenhahn", "scopus_author_id"=>"15061765600"}, {"first_name"=>"Julia", "last_name"=>"Albrecht", "scopus_author_id"=>"55668628600"}, {"first_name"=>"Antonio", "last_name"=>"Cosma", "scopus_author_id"=>"55880115100"}, {"first_name"=>"Ulrike", "last_name"=>"Protzer", "scopus_author_id"=>"55880004900"}, {"first_name"=>"Tanja", "last_name"=>"Bauer", "scopus_author_id"=>"10439345400"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"370004577", "doi"=>"10.1371/journal.pone.0076215", "sgr"=>"84885418048", "scopus"=>"2-s2.0-84885418048", "isbn"=>"1932-6203", "pmid"=>"24146841"}, "id"=>"be244a73-3d23-38c7-b1f2-956b8d25b95a", "abstract"=>"Polyfunctional CD4 or CD8 T cells are proposed to represent a correlate of immune control for persistent viruses as well as for vaccine mediated protection against infection. A well-suited methodology to study complex functional phenotypes of antiviral T cells is the combined staining of intracellular cytokines and phenotypic marker expression using polychromatic flow cytometry. In this study we analyzed the effect of an overnight resting period at 37 °C on the quantity and functionality of HIV-1, EBV, CMV, HBV and HCV specific CD4 and CD8 T-cell responses in a cohort of 21 individuals. We quantified total antigen specific T cells by multimer staining and used 10-color intracellular cytokine staining (ICS) to determine IFNγ, TNFα, IL2 and MIP1β production. After an overnight resting significantly higher numbers of functionally active T cells were detectable by ICS for all tested antigen specificities, whereas the total number of antigen specific T cells determined by multimer staining remained unchanged. Overnight resting shifted the quality of T-cell responses towards polyfunctionality and increased antigen sensitivity of T cells. Our data suggest that the observed effect is mediated by T cells rather than by antigen presenting cells. We conclude that overnight resting of PBMC prior to ex vivo analysis of antiviral T-cell responses represents an efficient method to increase sensitivity of ICS-based methods and has a prominent impact on the functional phenotype of T cells.", "link"=>"http://www.mendeley.com/research/overnight-resting-pbmc-changes-functional-signatures-antigen-specific-t-cell-responses-impact-immune", "reader_count"=>105, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Researcher"=>37, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>21, "Student > Postgraduate"=>9, "Other"=>10, "Student > Master"=>8, "Student > Bachelor"=>7, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Researcher"=>37, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>21, "Student > Postgraduate"=>9, "Other"=>10, "Student > Master"=>8, "Student > Bachelor"=>7, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>2, "Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>4, "Mathematics"=>1, "Agricultural and Biological Sciences"=>49, "Medicine and Dentistry"=>19, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Immunology and Microbiology"=>24}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>19}, "Neuroscience"=>{"Neuroscience"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>24}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>49}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Canada"=>2, "Belgium"=>1, "United States"=>1, "Japan"=>1, "Italy"=>1, "United Kingdom"=>3, "Australia"=>1, "Germany"=>1, "Spain"=>1}, "group_count"=>6}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1233798"], "description"=>"<p>(A) Flow cytometric analysis of lymphocyte viability compared between not rested and rested cells in a cohort of 21 individuals (** p<0.005, Wilcoxon matched pairs test; median indicated by black line). (B) Flow cytometric live/dead (NIR-/NIR+) discrimination and costaining with Annexin V for resolution between dead (NIR+) and apoptotic (NIR-/Annexin V+) cells in not rested and rested PBMC. NIR: near-infrared.</p>", "links"=>[], "tags"=>["resting", "post-thaw", "viability"], "article_id"=>820610, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_reduces_post_thaw_viability_of_PBMC_/820610", "title"=>"Overnight resting reduces post-thaw viability of PBMC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233799"], "description"=>"<p>(A) Comparison of total numbers of functional CD8 (left panel) and CD4 (right panel) T cells specific for HIV, HBV, HCV and EBV in a cohort of 21 individuals. (B) Single marker expression of antiviral CD8 (left panel) and CD4 (right panel) T cells. All analyses were performed on not rested and rested PBMC as indicated. Background values as determined in unstimulated controls are subtracted and a predefined threshold on subpopulation level is applied before calculating the total response and amount of single cytokines (see materials and methods). (*** p≤0.001; ** p≤0.05; * p≤0.05, Wilcoxon matched pairs test; median indicated by black line).</p>", "links"=>[], "tags"=>["resting", "antiviral", "t-cell", "responses", "detectable"], "article_id"=>820611, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_increases_the_magnitude_of_antiviral_T_cell_responses_detectable_by_ICS_/820611", "title"=>"Overnight resting increases the magnitude of antiviral T-cell responses detectable by ICS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233803"], "description"=>"<p>(A) HIV Nef specific CD8 (upper panel) and CD4 (lower panel) T cells were determined by ICS. Unstimulated PBMC served as negative controls (neg ctrl). Dot plots show functional T cells with numbers as% of total CD8 and CD4 T cells, respectively. Pie charts show the relative contribution of each T-cell subpopulation within the total HIV Nef specific T-cell response according to their functionality (mono- to poly-functional). One representative experiment is shown. (B) Pie charts represent the relative functional composition of total HIV, HBV, HCV and EBV specific CD8 (upper panel) and CD4 (lower panel) T-cell responses in a cohort of 21 individuals. All analyses were performed on not rested and rested PBMC as indicated.</p>", "links"=>[], "tags"=>["resting", "functionality", "antiviral"], "article_id"=>820613, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_increases_functionality_of_antiviral_T_cells_/820613", "title"=>"Overnight resting increases functionality of antiviral T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233804"], "description"=>"<p>(A) Bar charts indicate frequencies of bi- and mono-functional (blue: IFN-γ+/MIP-1β+; grey: MIP-1β+) HIV Nef specific CD8 T cells for a given peptide concentration (0.125–4 µg/ml) as percentage of total CD8 T cells. Cells were tested without (upper panel) or with (lower panel) overnight resting. (B) Pie charts show the functional profiles of HIV Nef specific CD8 T cells for a given peptide concentration. Cells were tested without (upper panel) or with (lower panel) overnight resting. (A) and (B) one representative experiment is shown. (C) Relative mean fluorescent intensity (rMFI) of IFN-γ staining of HIV Nef specific CD8 T cells of eight individuals normalized to rMFI of total CD3 T cells. All analyses were performed on not rested and rested PBMC as indicated. (** p<0.005, Wilcoxon matched pairs test; median indicated by black line).</p>", "links"=>[], "tags"=>["resting", "heightens"], "article_id"=>820614, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_heightens_sensitivity_to_antigens_/820614", "title"=>"Overnight resting heightens sensitivity to antigens.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233805"], "description"=>"<p>(A) Frequencies (left panel) and functionality (right panel) of HIV and CMV specific CD8 T cells were determined by ICS. Data indicate numbers of functional T cells as% of total CD8 T cells. Pie charts show the relative contribution of each functional subpopulation within the total CD8 T-cell response according to their functionality. (B) Frequencies of HIV and CMV specific CD8 T cells determined by multimer staining. Data indicate numbers of multimer-positive T cells as% of total CD8 T cells. (A) and (B): data from five HIV (n = 3) or CMV (n = 2) seropositive individuals are shown; median indicated by black line. (C) Comparison of T cell ICS and multimer staining. Bar charts show frequencies of functional epitope specific CD8 T cells determined by ICS calculated as percentage of corresponding multimer-positive T cells. Representative results of two individuals with known epitope specificities (HLA-B8 restricted HIV Nef and HLA-A2 restricted CMV IE1, respectively) are shown. All analyses were performed on not rested and rested PBMC as indicated.</p>", "links"=>[], "tags"=>["resting", "epitope"], "article_id"=>820615, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_does_not_alter_the_total_number_of_epitope_specific_T_cells_/820615", "title"=>"Overnight resting does not alter the total number of epitope specific T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233807"], "description"=>"<p>(A) Autologous EBV-transformed B-lymphoblastoid cell lines were used as antigen presenting cells to stimulate antigen specific T cells of an EBV seropositive subject. PBMC without the addition of EBV-transformed B-lymphoblastoid cell lines provided the negative controls used for background subtraction. (B) CD3 T cells of a CMV seropositive subject were isolated by magnetic cell sorting. After cryopreservation, CD3 T cells were stimulated directly or after overnight rest with a pool of overlapping peptides corresponding to the CMV-IE-1 protein. (A) and (B) IFNγ, IL2 and MIP1β production was determined by ICS directly or after an overnight resting. Frequencies (bar charts) and functional composition (pie charts) of EBV (A) and CMV (B) specific CD8 T cells are shown. CD8 T-cell subpopulations are depicted according to their functionality (three functions: green; two functions: blue; monofunctional cells: grey). One representative experiment is shown.</p>", "links"=>[], "tags"=>["mediated", "antigen", "presenting"], "article_id"=>820617, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resting_effect_is_not_mediated_by_antigen_presenting_cells_/820617", "title"=>"Resting effect is not mediated by antigen presenting cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233808"], "description"=>"<p>(A) Concentration of proinflammatory cytokines (TNFα, IL1β, IL6,) and chemokines (MIP1α, MIP1β, Rantes) in supernatants of unstimulated PBMC from five subjects were determined by cytometric bead array at indicated time points during the resting period of 18 hours. One representative experiment is shown.</p>", "links"=>[], "tags"=>["resting", "triggers", "proinflammatory", "cytokines"], "article_id"=>820618, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_triggers_production_of_proinflammatory_cytokines_and_chemokines_/820618", "title"=>"Overnight resting triggers production of proinflammatory cytokines and chemokines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233809"], "description"=>"<p>T-cell responses of an HIV infected patient were determined after whole blood stimulation and after stimulation of freshly isolated PBMC or cryopreserved PBMC of an HIV infected subject. IFNγ, IL2 and MIP1β production was determined by ICS. (A) Frequencies and (B) functional composition of HIV Nef specific CD8 T cells are shown. T-cell subpopulations are depicted according to their functionality (three functions: green; two functions: blue; monofunctional cells: grey). Analysis for (A) and (B) were performed on non-rested and rested PBMC as indicated. One representative experiment is shown.</p>", "links"=>[], "tags"=>["resting", "rescues", "cd8", "t-cell", "responses", "cryopreserved"], "article_id"=>820619, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_resting_rescues_CD8_T_cell_responses_of_cryopreserved_PBMC_/820619", "title"=>"Overnight resting rescues CD8 T-cell responses of cryopreserved PBMC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233810"], "description"=>"<p>Antibodies used for ICS of PBMC.</p>", "links"=>[], "tags"=>["ics"], "article_id"=>820620, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_used_for_ICS_of_PBMC_/820620", "title"=>"Antibodies used for ICS of PBMC.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233811"], "description"=>"<p>Antibodies used for ICS of whole blood.</p>", "links"=>[], "tags"=>["ics"], "article_id"=>820621, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_used_for_ICS_of_whole_blood_/820621", "title"=>"Antibodies used for ICS of whole blood.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-10-11 03:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/1233814", "https://ndownloader.figshare.com/files/1233815"], "description"=>"<div><p>Polyfunctional CD4 or CD8 T cells are proposed to represent a correlate of immune control for persistent viruses as well as for vaccine mediated protection against infection. A well-suited methodology to study complex functional phenotypes of antiviral T cells is the combined staining of intracellular cytokines and phenotypic marker expression using polychromatic flow cytometry. In this study we analyzed the effect of an overnight resting period at 37°C on the quantity and functionality of HIV-1, EBV, CMV, HBV and HCV specific CD4 and CD8 T-cell responses in a cohort of 21 individuals. We quantified total antigen specific T cells by multimer staining and used 10-color intracellular cytokine staining (ICS) to determine IFNγ, TNFα, IL2 and MIP1β production. After an overnight resting significantly higher numbers of functionally active T cells were detectable by ICS for all tested antigen specificities, whereas the total number of antigen specific T cells determined by multimer staining remained unchanged. Overnight resting shifted the quality of T-cell responses towards polyfunctionality and increased antigen sensitivity of T cells. Our data suggest that the observed effect is mediated by T cells rather than by antigen presenting cells. We conclude that overnight resting of PBMC prior to <i>ex vivo</i> analysis of antiviral T-cell responses represents an efficient method to increase sensitivity of ICS-based methods and has a prominent impact on the functional phenotype of T cells.</p></div>", "links"=>[], "tags"=>["resting", "pbmc", "signatures", "antigen", "t-", "monitoring"], "article_id"=>820624, "categories"=>["Biological Sciences"], "users"=>["Sarah Kutscher", "Claudia J. Dembek", "Simone Deckert", "Carolina Russo", "Nina Körber", "Johannes R. Bogner", "Fabian Geisler", "Andreas Umgelter", "Michael Neuenhahn", "Julia Albrecht", "Antonio Cosma", "Ulrike Protzer", "Tanja Bauer"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076215.s001", "https://dx.doi.org/10.1371/journal.pone.0076215.s002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overnight_Resting_of_PBMC_Changes_Functional_Signatures_of_Antigen_Specific_T_Cell_Responses_Impact_for_Immune_Monitoring_within_Clinical_Trials_/820624", "title"=>"Overnight Resting of PBMC Changes Functional Signatures of Antigen Specific T- Cell Responses: Impact for Immune Monitoring within Clinical Trials", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-10-11 03:16:34"}

PMC Usage Stats | Further Information

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Relative Metric

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