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pmc-enju-pas

bionlp-st-ge-2016-spacy-parsed

GO-BP

{"project":"GO-BP","denotations":[{"id":"T21963","span":{"begin":620,"end":629},"obj":"http://purl.obolibrary.org/obo/GO_0006915"},{"id":"T21964","span":{"begin":620,"end":629},"obj":"http://purl.obolibrary.org/obo/GO_0097194"},{"id":"T21965","span":{"begin":1052,"end":1072},"obj":"http://purl.obolibrary.org/obo/GO_0030154"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

GO-MF

{"project":"GO-MF","denotations":[{"id":"T21966","span":{"begin":802,"end":806},"obj":"http://purl.obolibrary.org/obo/GO_0005134"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T21967","span":{"begin":54,"end":59},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21968","span":{"begin":182,"end":187},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21969","span":{"begin":422,"end":427},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21970","span":{"begin":583,"end":588},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21971","span":{"begin":637,"end":642},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21972","span":{"begin":763,"end":768},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21973","span":{"begin":859,"end":864},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21974","span":{"begin":1193,"end":1198},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21975","span":{"begin":1301,"end":1306},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21976","span":{"begin":1337,"end":1342},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21977","span":{"begin":1491,"end":1496},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21978","span":{"begin":1609,"end":1614},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T21979","span":{"begin":1655,"end":1660},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

sentences

{"project":"sentences","denotations":[{"id":"T21238","span":{"begin":0,"end":28},"obj":"Sentence"},{"id":"T21239","span":{"begin":29,"end":151},"obj":"Sentence"},{"id":"T21240","span":{"begin":152,"end":277},"obj":"Sentence"},{"id":"T21241","span":{"begin":278,"end":397},"obj":"Sentence"},{"id":"T21242","span":{"begin":398,"end":552},"obj":"Sentence"},{"id":"T21243","span":{"begin":553,"end":619},"obj":"Sentence"},{"id":"T21244","span":{"begin":620,"end":701},"obj":"Sentence"},{"id":"T21245","span":{"begin":702,"end":839},"obj":"Sentence"},{"id":"T21246","span":{"begin":840,"end":1013},"obj":"Sentence"},{"id":"T21247","span":{"begin":1014,"end":1199},"obj":"Sentence"},{"id":"T21248","span":{"begin":1200,"end":1259},"obj":"Sentence"},{"id":"T21249","span":{"begin":1260,"end":1373},"obj":"Sentence"},{"id":"T21250","span":{"begin":1374,"end":1582},"obj":"Sentence"},{"id":"T21251","span":{"begin":1583,"end":1661},"obj":"Sentence"},{"id":"T294","span":{"begin":0,"end":28},"obj":"Sentence"},{"id":"T295","span":{"begin":29,"end":151},"obj":"Sentence"},{"id":"T296","span":{"begin":152,"end":277},"obj":"Sentence"},{"id":"T297","span":{"begin":278,"end":397},"obj":"Sentence"},{"id":"T298","span":{"begin":398,"end":552},"obj":"Sentence"},{"id":"T299","span":{"begin":553,"end":619},"obj":"Sentence"},{"id":"T300","span":{"begin":620,"end":701},"obj":"Sentence"},{"id":"T301","span":{"begin":702,"end":839},"obj":"Sentence"},{"id":"T302","span":{"begin":840,"end":1013},"obj":"Sentence"},{"id":"T303","span":{"begin":1014,"end":1199},"obj":"Sentence"},{"id":"T304","span":{"begin":1200,"end":1259},"obj":"Sentence"},{"id":"T305","span":{"begin":1260,"end":1373},"obj":"Sentence"},{"id":"T306","span":{"begin":1374,"end":1582},"obj":"Sentence"},{"id":"T307","span":{"begin":1583,"end":1661},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

events-check-again

{"project":"events-check-again","denotations":[{"id":"T21980","span":{"begin":35,"end":40},"obj":"Protein"},{"id":"T21981","span":{"begin":42,"end":45},"obj":"Protein"},{"id":"T21982","span":{"begin":47,"end":50},"obj":"Protein"},{"id":"T21983","span":{"begin":88,"end":91},"obj":"Protein"},{"id":"T21984","span":{"begin":92,"end":101},"obj":"Gene_expression"},{"id":"T21985","span":{"begin":109,"end":114},"obj":"Protein"},{"id":"T21986","span":{"begin":120,"end":123},"obj":"Protein"},{"id":"T21987","span":{"begin":141,"end":146},"obj":"Protein"},{"id":"T21988","span":{"begin":146,"end":149},"obj":"Protein"},{"id":"T21989","span":{"begin":159,"end":164},"obj":"Protein"},{"id":"T21990","span":{"begin":166,"end":168},"obj":"Protein"},{"id":"T21991","span":{"begin":170,"end":172},"obj":"Protein"},{"id":"T21992","span":{"begin":175,"end":178},"obj":"Protein"},{"id":"T21993","span":{"begin":224,"end":230},"obj":"Protein"},{"id":"T21994","span":{"begin":417,"end":420},"obj":"Protein"},{"id":"T21995","span":{"begin":441,"end":445},"obj":"Protein"},{"id":"T21996","span":{"begin":452,"end":461},"obj":"Protein"},{"id":"T21997","span":{"begin":503,"end":506},"obj":"Protein"},{"id":"T21998","span":{"begin":663,"end":672},"obj":"Protein"},{"id":"T21999","span":{"begin":802,"end":806},"obj":"Protein"},{"id":"T22000","span":{"begin":820,"end":823},"obj":"Protein"},{"id":"T22001","span":{"begin":824,"end":826},"obj":"Protein"},{"id":"T22002","span":{"begin":852,"end":855},"obj":"Protein"},{"id":"T22003","span":{"begin":991,"end":996},"obj":"Protein"},{"id":"T22004","span":{"begin":1001,"end":1006},"obj":"Protein"},{"id":"T22005","span":{"begin":1007,"end":1012},"obj":"Negative_regulation"},{"id":"T22006","span":{"begin":1007,"end":1012},"obj":"Negative_regulation"},{"id":"T22007","span":{"begin":1186,"end":1189},"obj":"Protein"},{"id":"T22008","span":{"begin":1294,"end":1297},"obj":"Protein"},{"id":"T22009","span":{"begin":1416,"end":1419},"obj":"Protein"},{"id":"T22010","span":{"begin":1597,"end":1600},"obj":"Protein"}],"relations":[{"id":"R17072","pred":"themeOf","subj":"T21983","obj":"T21984"},{"id":"R17073","pred":"themeOf","subj":"T22003","obj":"T22005"},{"id":"R17074","pred":"themeOf","subj":"T22004","obj":"T22006"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

bionlp-st-ge-2016-reference-tees

{"project":"bionlp-st-ge-2016-reference-tees","denotations":[{"id":"T22042","span":{"begin":29,"end":41},"obj":"Protein"},{"id":"T22043","span":{"begin":42,"end":45},"obj":"Protein"},{"id":"T22044","span":{"begin":47,"end":50},"obj":"Protein"},{"id":"T22045","span":{"begin":88,"end":91},"obj":"Protein"},{"id":"T22046","span":{"begin":109,"end":114},"obj":"Protein"},{"id":"T22047","span":{"begin":115,"end":119},"obj":"Protein"},{"id":"T22048","span":{"begin":120,"end":123},"obj":"Protein"},{"id":"T22049","span":{"begin":92,"end":101},"obj":"Gene_expression"},{"id":"T22050","span":{"begin":175,"end":179},"obj":"Protein"},{"id":"T22051","span":{"begin":441,"end":445},"obj":"Protein"},{"id":"T22052","span":{"begin":452,"end":457},"obj":"Protein"},{"id":"T22053","span":{"begin":458,"end":461},"obj":"Protein"},{"id":"T22054","span":{"begin":498,"end":510},"obj":"Protein"},{"id":"T22055","span":{"begin":802,"end":806},"obj":"Protein"},{"id":"T22056","span":{"begin":852,"end":856},"obj":"Protein"},{"id":"T22057","span":{"begin":991,"end":996},"obj":"Protein"},{"id":"T22058","span":{"begin":1001,"end":1012},"obj":"Protein"},{"id":"T22059","span":{"begin":1186,"end":1190},"obj":"Protein"},{"id":"T22060","span":{"begin":1200,"end":1205},"obj":"Protein"},{"id":"T22061","span":{"begin":1294,"end":1298},"obj":"Protein"},{"id":"T22062","span":{"begin":1331,"end":1336},"obj":"Protein"},{"id":"T22063","span":{"begin":1411,"end":1423},"obj":"Protein"},{"id":"T22064","span":{"begin":1597,"end":1601},"obj":"Protein"},{"id":"T22065","span":{"begin":1640,"end":1644},"obj":"Protein"},{"id":"T22066","span":{"begin":1649,"end":1654},"obj":"Protein"},{"id":"T22067","span":{"begin":1627,"end":1636},"obj":"Gene_expression"},{"id":"T22068","span":{"begin":1627,"end":1636},"obj":"Gene_expression"},{"id":"T22069","span":{"begin":1615,"end":1622},"obj":"Negative_regulation"},{"id":"T22070","span":{"begin":1615,"end":1622},"obj":"Negative_regulation"}],"relations":[{"id":"R17078","pred":"themeOf","subj":"T22045","obj":"T22049"},{"id":"R17079","pred":"themeOf","subj":"T22065","obj":"T22067"},{"id":"R17080","pred":"themeOf","subj":"T22066","obj":"T22068"},{"id":"R17081","pred":"themeOf","subj":"T22067","obj":"T22069"},{"id":"R17082","pred":"themeOf","subj":"T22068","obj":"T22070"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

bionlp-st-ge-2016-reference

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T21553","span":{"begin":35,"end":40},"obj":"Q9BZS1"},{"id":"T21554","span":{"begin":42,"end":45},"obj":"P01730"},{"id":"T21555","span":{"begin":47,"end":50},"obj":"P01732"},{"id":"T21556","span":{"begin":47,"end":50},"obj":"P10966"},{"id":"T21557","span":{"begin":88,"end":91},"obj":"Q9U6Y4"},{"id":"T21558","span":{"begin":109,"end":114},"obj":"Q9BZS1"},{"id":"T21559","span":{"begin":120,"end":123},"obj":"Q9U6Y4"},{"id":"T21560","span":{"begin":175,"end":178},"obj":"P01730"},{"id":"T21561","span":{"begin":224,"end":228},"obj":"P08575"},{"id":"T21562","span":{"begin":417,"end":420},"obj":"P01730"},{"id":"T21563","span":{"begin":441,"end":445},"obj":"P01589"},{"id":"T21564","span":{"begin":452,"end":457},"obj":"Q9BZS1"},{"id":"T21565","span":{"begin":458,"end":461},"obj":"Q9U6Y4"},{"id":"T21566","span":{"begin":503,"end":506},"obj":"P20963"},{"id":"T21567","span":{"begin":503,"end":506},"obj":"P07766"},{"id":"T21568","span":{"begin":503,"end":506},"obj":"P04234"},{"id":"T21569","span":{"begin":503,"end":506},"obj":"P09693"},{"id":"T21570","span":{"begin":663,"end":672},"obj":"P08758"},{"id":"T21571","span":{"begin":802,"end":806},"obj":"P60568"},{"id":"T21572","span":{"begin":820,"end":823},"obj":"P07766"},{"id":"T21573","span":{"begin":820,"end":823},"obj":"P04234"},{"id":"T21574","span":{"begin":820,"end":823},"obj":"P09693"},{"id":"T21575","span":{"begin":820,"end":823},"obj":"P20963"},{"id":"T21576","span":{"begin":852,"end":855},"obj":"P01730"},{"id":"T21577","span":{"begin":991,"end":996},"obj":"Q01196"},{"id":"T21578","span":{"begin":1001,"end":1006},"obj":"Q13761"},{"id":"T21579","span":{"begin":1186,"end":1189},"obj":"P01730"},{"id":"T21580","span":{"begin":1294,"end":1297},"obj":"P01730"},{"id":"T21581","span":{"begin":1416,"end":1419},"obj":"P04234"},{"id":"T21582","span":{"begin":1416,"end":1419},"obj":"P07766"},{"id":"T21583","span":{"begin":1416,"end":1419},"obj":"P09693"},{"id":"T21584","span":{"begin":1416,"end":1419},"obj":"P20963"},{"id":"T21585","span":{"begin":1597,"end":1600},"obj":"P01730"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

test2

{"project":"test2","denotations":[{"id":"T21180","span":{"begin":35,"end":40},"obj":"Protein"},{"id":"T21181","span":{"begin":42,"end":45},"obj":"Protein"},{"id":"T21182","span":{"begin":47,"end":50},"obj":"Protein"},{"id":"T21183","span":{"begin":88,"end":91},"obj":"Protein"},{"id":"T21184","span":{"begin":92,"end":101},"obj":"Gene_expression"},{"id":"T21185","span":{"begin":109,"end":114},"obj":"Protein"},{"id":"T21186","span":{"begin":120,"end":123},"obj":"Protein"},{"id":"T21187","span":{"begin":141,"end":149},"obj":"Protein"},{"id":"T21188","span":{"begin":159,"end":172},"obj":"Protein"},{"id":"T21189","span":{"begin":175,"end":178},"obj":"Protein"},{"id":"T21190","span":{"begin":224,"end":230},"obj":"Protein"},{"id":"T21191","span":{"begin":417,"end":420},"obj":"Protein"},{"id":"T21192","span":{"begin":441,"end":445},"obj":"Protein"},{"id":"T21193","span":{"begin":452,"end":461},"obj":"Protein"},{"id":"T21194","span":{"begin":503,"end":506},"obj":"Protein"},{"id":"T21195","span":{"begin":663,"end":672},"obj":"Protein"},{"id":"T21196","span":{"begin":802,"end":806},"obj":"Protein"},{"id":"T21197","span":{"begin":820,"end":823},"obj":"Protein"},{"id":"T21198","span":{"begin":824,"end":826},"obj":"Protein"},{"id":"T21199","span":{"begin":852,"end":855},"obj":"Protein"},{"id":"T21200","span":{"begin":991,"end":996},"obj":"Protein"},{"id":"T21201","span":{"begin":1001,"end":1006},"obj":"Protein"},{"id":"T21202","span":{"begin":1007,"end":1012},"obj":"Negative_regulation"},{"id":"T21203","span":{"begin":1186,"end":1189},"obj":"Protein"},{"id":"T21204","span":{"begin":1294,"end":1297},"obj":"Protein"},{"id":"T21205","span":{"begin":1416,"end":1419},"obj":"Protein"},{"id":"T21206","span":{"begin":1597,"end":1600},"obj":"Protein"}],"relations":[{"id":"R16425","pred":"themeOf","subj":"T21183","obj":"T21184"},{"id":"R16426","pred":"themeOf","subj":"T21200","obj":"T21202"},{"id":"R16427","pred":"themeOf","subj":"T21201","obj":"T21202"}],"text":"In vitro suppression assays.\nMouse Foxp3+ CD4+ CD8− T cells were FACS purified based on GFP expressed from a Foxp3-IRES-GFP knock-in allele (Foxp3GFP). Naive (CD62Lhi44lo25−) CD4+ T cells (effectors) were FACS-purified from Cd45.1 mice, then loaded with 5 µM CFSE (Invitrogen). Total splenocytes from C57BL/6 mice inactivated with 50 µg/ml mitomycin C (Sigma-Aldrich) for 45 min were used as APCs. A total of 4 × 105 CD4+ cells (CFSE-loaded CD25− plus Foxp3-GFP+) was mixed with 105 APCs + 1 µg/ml anti-CD3 mAb per well of a 96 well round bottom plate. Proliferation of the effector cells was analyzed by CFSE dilution. Apoptosis of the cells was investigated by annexin V staining and flow cytometry. Positive and negative control gates were made according to T cells cultured only in the presence of IL-2 without anti-CD3/28 stimulation.\nHuman naive CD4+ T cells were isolated by negative selection by MACS from PBMCs and either transfected with a scrambled siRNA or with a combination of RUNX1 and RUNX3 siRNA. Cells were then cultured under iT reg cell differentiating conditions and mixed with 2 × 105 autologous irradiated PBMCs that were used as APCs and autologous CFSE-labeled CD4+ T cells. T reg cell to responder cell ratio was 1:20, 1:10, and 1:5. To check the proliferation of the CD4+ T cells without suppression, no T reg cells were added in a control group. Cells were stimulated with 2.5 µg/ml anti-CD3 mAb, cultured in a 96-well plate and the proliferation of the effector cells was determined by analyzing the CFSE dilution by flow cytometry after 5 d of culture. Gating on the CD4+CFSE+ T cells enabled the exclusion of APCs and T reg cells."}

PMC:2806624 / 42890-44551 JSONTXT

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PMC:2806624 / 42890-44551 JSON TXT