A Quantitative Study of the Mechanisms behind Thymic Atrophy in Gαi2-Deficient Mice during Colitis Development
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{"title"=>"A quantitative study of the mechanisms behind thymic atrophy in gαi2-deficient mice during colitis development", "type"=>"journal", "authors"=>[{"first_name"=>"Kristina", "last_name"=>"Elgbratt", "scopus_author_id"=>"21734218100"}, {"first_name"=>"Andreas", "last_name"=>"Jansson", "scopus_author_id"=>"8584517900"}, {"first_name"=>"Elisabeth", "last_name"=>"Hultgren-Hörnquist", "scopus_author_id"=>"7801398639"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84861002198", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0036726", "pmid"=>"22590596", "pui"=>"364803024", "scopus"=>"2-s2.0-84861002198"}, "id"=>"e89b6052-d921-3e2e-814d-a695964608da", "abstract"=>"Mice deficient for the G protein subunit Gαi2 spontaneously develop colitis, a chronic inflammatory disease associated with dysregulated T cell responses. We and others have previously demonstrated a thymic involution in these mice and an aberrant thymocyte dynamics. The Gαi2(-/-) mice have a dramatically reduced fraction of double positive thymocytes and an increased fraction of single positive (SP) thymocytes. In this study, we quantify a number of critical parameters in order to narrow down the underlying mechanisms that cause the dynamical changes of the thymocyte development in the Gαi2(-/-) mice. Our data suggest that the increased fraction of SP thymocytes results only from a decreased number of DP thymocytes, since the number of SP thymocytes in the Gαi2(-/-) mice is comparable to the control littermates. By measuring the frequency of T cell receptor excision circles (TRECs) in the thymocytes, we demonstrate that the number of cell divisions the Gαi2(-/-) SP thymocytes undergo is comparable to SP thymocytes from control littermates. In addition, our data show that the mature SP CD4(+) and CD8(+) thymocytes divide to the same extent before they egress from the thymus. By estimating the number of peripheral TREC(+) T lymphocytes and their death rate, we could calculate the daily egression of thymocytes. Gαi2(-/-) mice with no/mild and moderate colitis were found to have a slower export rate in comparison to the control littermates. The quantitative measurements in this study suggest a number of dynamical changes in the thymocyte development during the progression of colitis.", "link"=>"http://www.mendeley.com/research/quantitative-study-mechanisms-behind-thymic-atrophy-g%CE%B1i2deficient-mice-during-colitis-development", "reader_count"=>3, "reader_count_by_academic_status"=>{"Student > Ph. D. Student"=>2, "Student > Master"=>1}, "reader_count_by_user_role"=>{"Student > Ph. D. Student"=>2, "Student > Master"=>1}, "reader_count_by_subject_area"=>{"Agricultural and Biological Sciences"=>2, "Medicine and Dentistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/640554"], "description"=>"<p>Peripheral blood was collected and the volume recorded from individual mice, whereafter Gαi2<sup>+/−</sup> mice were pooled (3 mice/pool) and analysed, while peripheral blood from Gαi2<sup>−/−</sup> mice was analysed individually. The control group of wt mice were 4–9 weeks old (n = 11–16, white circles) and the Gαi2<sup>−/−</sup> mice were 4–8 weeks old (n = 4–14, gray/black symbols), grouped based on their colitis scores (no colitis = rectangles, mild colitis = triangles). PBMCs were isolated from peripheral blood and the frequencies of CD3<sup>+</sup> T lymphocytes was analysed by flow cytometry. The number of CD3<sup>+</sup> (A) and naïve T lymphocytes (B) are presented as total T lymphocytes of the entire blood volume in each mouse, 6–8 ml blood per 100 g body weight <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036726#pone.0036726-Hoff1\" target=\"_blank\">[40]</a>. (C) The frequencies of TREC<sup>+</sup> lymphocytes were analysed by real time-PCR on isolated DNA from flow cytometry sorted peripheral blood CD3<sup>+</sup> T lymphocytes. (D) Numbers of TREC<sup>+</sup> CD3<sup>+</sup> peripheral blood lymphocytes, as calculated from data (A) and (C). Horizontal lines indicate the mean within the group. Statistical analysis was performed using the Mann-Whitney non-parametric test and values of p≤0.05 were considered significant. *p≤0.05; **p≤0.01 and ***p≤0.001 between Gαi2<sup>−/−</sup> and wt mice.</p>", "links"=>[], "tags"=>["frequencies", "numbers", "trecs", "peripheral", "lymphocytes", "colitic"], "article_id"=>311040, "categories"=>["Chemistry", "Immunology"], "users"=>["Kristina Elgbratt", "Andreas Jansson", "Elisabeth Hultgren-Hörnquist"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036726.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decreased_frequencies_and_numbers_of_TRECs_in_peripheral_blood_CD3_T_lymphocytes_in_colitic_G_945_i2_8722_8722_mice_/311040", "title"=>"Decreased frequencies and numbers of TRECs in peripheral blood CD3<sup>+</sup> T lymphocytes in colitic Gαi2<sup>−/−</sup> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:22:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/640435"], "description"=>"<p>Mature CD4<sup>+</sup>CD8<sup>−</sup>CD62L<sup>+</sup>CD69<sup>−</sup> and CD8<sup>+</sup>CD4<sup>−</sup>CD62L<sup>+</sup>CD69<sup>−</sup> thymocytes from wt mice and Gαi2<sup>−/−</sup> mice were sorted by flow cytometry. DNA from the sorted thymocytes were analysed for the amount of TRECs specific DNA by real time-PCR. The frequencies of TRECs<sup>+</sup> cells in CD4<sup>+</sup>CD62L<sup>+</sup> (white symbols) and CD8<sup>+</sup>CD62L<sup>+</sup> (gray symbols) thymocytes were analysed in 4–9 weeks old wt mice (n = 28) and 4–8 weeks old Gαi2<sup>−/−</sup> mice (n = 5–13), grouped based on colitis scores (rectangles = no colitis, triangles = mild colitis). The mean frequencies within each group are represented as a horizontal line. Statistical analysis was performed using the Mann-Whitney non-parametric test and values of p≤0.05 were considered significant.</p>", "links"=>[], "tags"=>["frequencies", "trecs", "mature", "thymocytes", "mice", "colitis"], "article_id"=>310925, "categories"=>["Chemistry", "Immunology"], "users"=>["Kristina Elgbratt", "Andreas Jansson", "Elisabeth Hultgren-Hörnquist"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036726.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_frequencies_of_TRECs_in_mature_thymocytes_in_G_i2_8722_8722_mice_are_independent_of_the_colitis_score_/310925", "title"=>"The frequencies of TRECs in mature thymocytes in Gαi2<sup>−/−</sup> mice are independent of the colitis score.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:21:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/640352"], "description"=>"<p>Flow cytometric analyses of the frequencies and total numbers of CD4<sup>+</sup>8<sup>+</sup> (DP) (A–B), CD4<sup>+</sup>8<sup>−</sup> (CD4 SP) (C–D) and CD4<sup>−</sup>8<sup>+</sup> (CD8 SP) (E–F) thymocytes. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036726#s3\" target=\"_blank\">Results</a> from wt mice (n = 28–31) and Gαi2<sup>−/−</sup> mice (n = 6–14) are shown as frequencies or numbers of thymocytes from individual mice with the mean within each group presented as a horizontal line. The control group consisted of 4–9 weeks old wt mice whereas the Gαi2<sup>−/−</sup> mice were 4–8 weeks old, grouped based on their colitis score; no and mild colitis (rectangles and triangles, respectively), moderate (dark gray circles) and severe colitis (black circles). Statistical analysis was performed using the Mann-Whitney non-parametric test and values of p≤0.05 were considered significant, *p≤0.05; **p≤0.01 and ***p≤0.001 between Gαi2<sup>−/−</sup> mice and control mice or between Gαi2<sup>−/−</sup> mice with different colitis scores.</p>", "links"=>[], "tags"=>["numbers", "frequencies", "dp", "thymocytes", "sp", "mice", "compared"], "article_id"=>310830, "categories"=>["Chemistry", "Immunology"], "users"=>["Kristina Elgbratt", "Andreas Jansson", "Elisabeth Hultgren-Hörnquist"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036726.g001", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decreased_numbers_and_frequencies_of_DP_thymocytes_but_equal_numbers_of_SP_thymocytes_in_G_i2_8722_8722_mice_compared_to_control_mice_/310830", "title"=>"Decreased numbers and frequencies of DP thymocytes but equal numbers of SP thymocytes in Gαi2<sup>−/−</sup> mice compared to control mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:21:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/640689"], "description"=>"a<p>The different age groups were: 4 (n = 9), 5 (n = 7), 6 (n = 5), 7 (n = 6) and 8 weeks (n = 3).</p>b<p>The different colitis score were: 0p (n = 3), 0.5–2p (n = 3), 2.5–4p (n = 14), 4.5–6p (n = 6) and 6.5–8p (n = 4).</p>", "links"=>[], "tags"=>["colitis", "thymocyte"], "article_id"=>311185, "categories"=>["Chemistry", "Immunology"], "users"=>["Kristina Elgbratt", "Andreas Jansson", "Elisabeth Hultgren-Hörnquist"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036726.t001", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Spearman_s_correlation_analysis_between_age_or_colitis_score_and_fraction_or_number_of_the_different_thymocyte_subpopulations_/311185", "title"=>"Spearman's correlation analysis between age or colitis score and fraction or number of the different thymocyte subpopulations.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:23:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/640625"], "description"=>"<p>A. PBMCs were stained with CD4 and CD62L specific antibodies, as well as Annexin V and Propidium Iodide (PI) to analyse the frequency of naïve peripheral blood CD4<sup>+</sup> T lymphocytes undergoing apoptosis. The bars depicts the mean frequency ± S.D. of apoptotic CD4<sup>+</sup> T lymphocytes in wt mice (n = 6, light bar) and Gαi2<sup>−/−</sup> mice (n = 5, dark bar (A). The daily egression was calculated by the following equation; , see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036726#s2\" target=\"_blank\">Materials and Methods</a>. (B) Four-nine weeks old wt mice (n = 11) and Gαi2<sup>−/−</sup> mice (n = 4–11) indicate the individual numbers of thymocytes egressing per day. Gαi2<sup>−/−</sup> mice were divided based on their stage of colitis (no colitis = rectangles, mild colitis = triangles). Horizontal lines indicate mean values within each group. Statistical analysis was performed using the Mann-Whitney non-parametric test and values of p≤0.05 were considered significant. *p≤0.05; **p≤0.01 and ***p≤0.001 between Gαi2<sup>−/−</sup> and wt mice.</p>", "links"=>[], "tags"=>["thymic", "egression", "mice"], "article_id"=>311120, "categories"=>["Chemistry", "Immunology"], "users"=>["Kristina Elgbratt", "Andreas Jansson", "Elisabeth Hultgren-Hörnquist"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036726.g004", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reduced_thymic_egression_in_G_i2_8722_8722_mice_with_no_mild_and_moderate_colitis_/311120", "title"=>"Reduced thymic egression in Gαi2<sup>−/−</sup> mice with no/mild and moderate colitis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:22:45"}

PMC Usage Stats | Further Information

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Relative Metric

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