The Shedding of CD62L (L-Selectin) Regulates the Acquisition of Lytic Activity in Human Tumor Reactive T Lymphocytes
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{"title"=>"The shedding of CD62L (L-selectin) regulates the acquisition of lytic activity in human tumor reactive T lymphocytes", "type"=>"journal", "authors"=>[{"first_name"=>"Shicheng", "last_name"=>"Yang", "scopus_author_id"=>"35217051700"}, {"first_name"=>"Fang", "last_name"=>"Liu", "scopus_author_id"=>"57091817700"}, {"first_name"=>"Qiong J.", "last_name"=>"Wang", "scopus_author_id"=>"8835440900"}, {"first_name"=>"Steven A.", "last_name"=>"Rosenberg", "scopus_author_id"=>"55572006200"}, {"first_name"=>"Richard A.", "last_name"=>"Morgan", "scopus_author_id"=>"7403329955"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-79960884563", "sgr"=>"79960884563", "pui"=>"362231750", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"21829468", "doi"=>"10.1371/journal.pone.0022560"}, "id"=>"40fe53ea-b98c-358c-b073-d3141f326d1e", "abstract"=>"CD62L/L-selectin is a marker found on naïve T cells and further distinguishes central memory (Tcm, CD62L+) from effector memory (Tem, CD62L-) T cells. The regulation of CD62L plays a pivotal role in controlling the traffic of T lymphocytes to and from peripheral lymph nodes. CD62L is shed from the cell membrane following T cell activation, however, the physiological significance of this event remains to be elucidated. In this study, we utilized in vitro generated anti-tumor antigen T cells and melanoma lines as a model to evaluate the dynamics of CD62L shedding and expression of CD107a as a marker of lytic activity. Upon encounter, with matched tumor lines, antigen reactive T cells rapidly lose CD62L expression and this was associated with the acquisition of CD107a. By CD62L ELISA, we confirmed that this transition was mediated by the shedding of CD62L when T cells encountered specific tumor antigen. The introduction of a shedding resistant mutant of CD62L into the tumor antigen-reactive T cell line JKF6 impaired CD107a acquisition following antigen recognition and this was correlated with decreased lytic activity as measured by (51)Cr release assays. The linkage of the shedding of CD62L from the surface of anti-tumor T cells and acquisition of lytic activity, suggests a new function for CD62L in T cell effector functions and anti-tumor activity.", "link"=>"http://www.mendeley.com/research/shedding-cd62l-lselectin-regulates-acquisition-lytic-activity-human-tumor-reactive-t-lymphocytes", "reader_count"=>80, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>13, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>34, "Student > Postgraduate"=>3, "Student > Master"=>11, "Other"=>2, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>13, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>34, "Student > Postgraduate"=>3, "Student > Master"=>11, "Other"=>2, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>8, "Agricultural and Biological Sciences"=>42, "Medicine and Dentistry"=>9, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Chemistry"=>1, "Immunology and Microbiology"=>15}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>9}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>15}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>42}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"Netherlands"=>1, "United States"=>2, "United Kingdom"=>2}, "group_count"=>5}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/751599"], "description"=>"<p><b>A.</b> The dynamics of CD62L shedding in CD8+/CD62L+ population was followed over 6hr. CD8+ MART-1 TCR transduced Tcm cells were sorted by FACS as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0022560#pone-0022560-g003\" target=\"_blank\">figure 3</a>. These sorted cells (CD45RO+/CD62L+) were co-cultured with melanoma lines 526 (HLA-A0201+) and 938 (HLA-A0201−) for the times indicated and analyzed for CD45RO and CD62L. Column on left (co-cultured with 938) and on right (co-cultured with 526). The numbers in each quadrant represent the percent of cells in that quadrant. <b>B.</b> The expression of CD107a correlates with the shedding of CD62L in CD8+/CD62L+ population. CD8+ MART-1 TCR transduced T cells from a different donor were co-cultures as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0022560#s2\" target=\"_blank\">methods</a>. The cells were gated on CD3/CD8/MART-1+ and then plotted for CD107a and CD62L expression. The samples were collected and stained at the times indicated on right. The fluorophores conjugated antibodies for this analysis were CD107a FITC, MART-1 PE, CD62L APC, CD45RO PE-Cy7, CD8 APC-Cy7 and propidium iodide staining.</p>", "links"=>[], "tags"=>["cd62l", "shedding", "inversely", "correlated"], "article_id"=>421968, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g004", "stats"=>{"downloads"=>1, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_dynamics_of_CD62L_shedding_is_inversely_correlated_with_the_expression_of_CD107a_/421968", "title"=>"The dynamics of CD62L shedding is inversely correlated with the expression of CD107a.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:17:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/751489"], "description"=>"<p><b>Top.</b> MART-1 TCR transduced cells were gated on the CD8+ and CD45RO+/CD62L+ (Tcm) population and sorted by FACS. The sorted Tcm cells were co-cultured with melanoma lines for 4 h. Culture media (supernatant) was collected and the cells stained for FACS analysis. Shown in the top plots is the expression of CD45RO and CD62L in unmanipulated Tcm cells, Tcm cells cultured with melanoma line 938 (HLA-A0201−), and Tcm cells cultured with melanoma line 526 (HLA-A0201+). The numbers in each quadrant represent the percent of cells in that quadrant. The fluorophore conjugated antibodies used in this analysis were CD107a FITC, CD62L PE, CD45RO APC, CD3 APC-Cy7 with propidium iodide staining. <b>Bottom.</b> The amount of CD62L in the cell media supernatant and total cell lysates was measured by ELISA. The amount of CD62L in the total cellular lysate in the Tcm group was set as 100% for comparison to the other lysate values (left side bars). For the cell supernatant determinations, the amount of soluble CD62L in the supernatant from the Tcm+526 group was set as 100%, for comparison to other values (right side bars). Data shown are the normalized values ± stdev for three independent determinations.</p>", "links"=>[], "tags"=>["cd62l", "antigen", "induced", "shedding"], "article_id"=>421859, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_loss_of_CD62L_is_due_to_antigen_induced_shedding_in_T_cells_/421859", "title"=>"The loss of CD62L is due to antigen induced shedding in T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:17:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/751745"], "description"=>"<p><b>A.</b> TIL line JKF6 was transduced with lentiviral vector harboring human CD62L and its shedding resistant mutant dK-S. JKF6 cells expressing CD62L and the dK-S mutant were enriched using CD62L beads and analyzed for CD62L expression by FACS. <b>B.</b> JKF6 gene engineered cell lines were co-cultured with melanoma lines 526 (HLA-A0201+) and 938 (HLA-A0201−) analyzed for CD45RO and CD62L expression (Mock, mock transduced cells; WT, CD62L transduced JKF6; dK-S, JKF6 transduced with shedding resistant mutant). On left, FACS plots from co-cultures of different groups; on right, overlayed histograms of the expression of CD107a in Tem and Tcm subsets. Solid line (CD45RO+CD62L−) indicates Tem, semi-solid line (CD45RO+CD62L+) indicates Tcm. <b>C.</b> Quantification of CD107a expression from triplicate determinations was calculated and plotted as % CD107a+ (mean ± STDEV). Statistical analysis using t test; *: dK-S vs mock and WT, p<0.01. <b>D.</b> The lytic activity of JKF6 lines expressing CD62L and its mutant were determined by co-culture with <sup>51</sup>Cr-labeled melanoma lines at the indicated ratios. The percent cell lysis was calculated using the formula ((specific release-spontaneous release)/(total release-spontaneous release))×100. Results representing mean of triplicate cultures were plotted. *: dK-S vs WT, p<0.05; #: dK-S vs Mock, p<0.05. The t-Test with two samples assuming equal variances was used for statistical analysis. The fluorophores conjugated antibodies for this analysis were CD107a FITC, MART-1 PE, CD62L APC, CD45RO PE-Cy7, CD8 APC-Cy7 and propidium iodide staining.</p>", "links"=>[], "tags"=>["shedding", "resistant", "cd62l", "affects"], "article_id"=>422116, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g005", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Introduction_of_shedding_resistant_CD62L_affects_the_surface_expression_of_CD107a_/422116", "title"=>"Introduction of shedding resistant CD62L affects the surface expression of CD107a.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:18:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/751331"], "description"=>"<p><b>A.</b> TCR transduced cells were cultured as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0022560#s2\" target=\"_blank\">methods</a>. Shown is the FACS analysis, where cells were gated on CD3/CD8/MART-1+ or CD3/CD8/MART-1−, and then plotted using markers CD45RO and CD62L (left two column plots). The expression of CD107a on each group was then determined and separately plotted in the histograms on right corresponding to column data on left. In each histogram the expression of CD107a on each subset was overlaid reflecting Tem, Tcm and naïve populations. Red (CD45RO+CD62L−) indicates Tem, blue (CD45RO+CD62L+) indicates Tcm, and green (CD45RO−CD62L+) indicates Tn population. <b>B.</b> The cells were processed as described above. By FACS analysis, the cells were gated on CD3/CD8/MART-1+ or CD3/CD8/MART-1−, then plotted using markers CD107a and CD62L in dot plots. The fluorophore conjugated antibodies used in this analysis were CD107a FITC, CD62L PE, CD45RO APC, CD3 APC-Cy7 with propidium iodide staining.</p>", "links"=>[], "tags"=>["cd62l", "antigen", "cd107a", "occurs"], "article_id"=>421701, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g002", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_loss_of_CD62L_is_tumor_antigen_specific_and_the_expression_of_CD107a_occurs_on_CD62L_negative_cells_/421701", "title"=>"The loss of CD62L is tumor antigen specific and the expression of CD107a occurs on CD62L negative cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:16:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/751177"], "description"=>"<p><b>A.</b> MART-1 TCR vector-transduced T cells were co-cultured with melanoma lines 526, 624 (both HLA-A*0201 positive) and 938 (HLA-A*0201 negative) for four hours and then analyzed by FACS. The first column on left, the cells were plotted using differentiation markers CD45RO and CD62L into 4 subsets, the expression of CD107a on each subset was denoted in the histograms to the right. <b>B.</b> Based on data from three independent experiments, the expression of CD107a after co-culture with melanoma lines was determined and mean% ± stdev plotted. Student t-test was used for statistical analysis. The fluorophore conjugated antibodies used in this analysis were CD107a FITC, CD62L PE, CD45RO APC, CD3 APC-Cy7 with propidium iodide staining.</p>", "links"=>[], "tags"=>["cd62l", "correlates", "cd107a"], "article_id"=>421542, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g001", "stats"=>{"downloads"=>3, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_loss_of_CD62L_is_specific_and_correlates_with_CD107a_surface_expression_/421542", "title"=>"The loss of CD62L is specific and correlates with CD107a surface expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:15:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/751862"], "description"=>"<p><b>A.</b> JKF6 lines expressing CD62L and mutant dK-S were activated using PMA/Ionomycin (PMA/Ion) for 4 h, and the surface expression for CD107a and IFN-γ was evaluated by FACS. One representative result from three independent experiments is shown (Mock, mock transduced cells; WT, CD62L transduced JKF6; dK-S, JKF6 transduced with shedding resistant mutant). For CD107a live cell staining, the fluorophores conjugated antibodies were CD107a FITC, CD62L APC, CD3 PE and propidium iodide staining; For intracellular staining of IFN-γ, the fluorophore conjugated antibodies were IFN-γ FITC, CD62L APC, CD3 PE, without propidium iodide staining. <b>B.</b> Plot of the percent positive cells for CD107a and IFN-γ on the surface of JKF6 lines expressing wild type and shedding resistant CD62L mutant. The mean ± STDEV from three independent experiments is displayed, and t-test was used for statistical analysis.</p>", "links"=>[], "tags"=>["shedding", "resistant", "cd62l", "jkf6", "hinders", "cd107a", "non-specific"], "article_id"=>422223, "categories"=>["Cancer", "Immunology"], "users"=>["Shicheng Yang", "Fang Liu", "Qiong J. Wang", "Steven A. Rosenberg", "Richard A. Morgan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022560.g006", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Introduction_of_shedding_resistant_CD62L_into_JKF6_hinders_CD107a_expression_following_non_specific_activation_/422223", "title"=>"Introduction of shedding resistant CD62L into JKF6 hinders CD107a expression following non-specific activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:19:10"}

PMC Usage Stats | Further Information

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