PMC:1064873 / 8204-9280 JSON TXT

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LappsTest

{"project":"LappsTest","denotations":[{"id":"T7328","span":{"begin":1006,"end":1010},"obj":"Protein"},{"id":"T7327","span":{"begin":533,"end":538},"obj":"Protein"},{"id":"T7326","span":{"begin":517,"end":528},"obj":"Protein"},{"id":"T7325","span":{"begin":383,"end":387},"obj":"Protein"},{"id":"T7324","span":{"begin":264,"end":272},"obj":"Protein"},{"id":"T7323","span":{"begin":199,"end":203},"obj":"Protein"},{"id":"T7322","span":{"begin":13,"end":17},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

2_test

{"project":"2_test","denotations":[{"id":"15535835-9182920-4156886","span":{"begin":452,"end":454},"obj":"9182920"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

biosemtest

pmc-enju-pas

bionlp-st-ge-2016-test-proteins

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T7482","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7481","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7480","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7479","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7478","span":{"begin":13,"end":16},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T7493","span":{"begin":1006,"end":1009},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7492","span":{"begin":383,"end":386},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7491","span":{"begin":269,"end":272},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7490","span":{"begin":199,"end":202},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7489","span":{"begin":13,"end":16},"obj":"http://www.uniprot.org/uniprot/P01730"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

UBERON-AE

{"project":"UBERON-AE","denotations":[{"id":"T7341","span":{"begin":574,"end":580},"obj":"http://purl.obolibrary.org/obo/UBERON_0000479"},{"id":"T7340","span":{"begin":99,"end":104},"obj":"http://purl.obolibrary.org/obo/UBERON_0000178"},{"id":"T7339","span":{"begin":37,"end":42},"obj":"http://purl.obolibrary.org/obo/UBERON_0000178"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T7488","span":{"begin":1013,"end":1018},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7487","span":{"begin":981,"end":986},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7486","span":{"begin":390,"end":395},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7485","span":{"begin":206,"end":211},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7484","span":{"begin":55,"end":60},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7483","span":{"begin":20,"end":25},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

sentences

{"project":"sentences","denotations":[{"id":"T7472","span":{"begin":968,"end":1076},"obj":"Sentence"},{"id":"T7471","span":{"begin":457,"end":967},"obj":"Sentence"},{"id":"T7470","span":{"begin":383,"end":456},"obj":"Sentence"},{"id":"T7469","span":{"begin":199,"end":382},"obj":"Sentence"},{"id":"T7468","span":{"begin":26,"end":198},"obj":"Sentence"},{"id":"T7467","span":{"begin":0,"end":25},"obj":"Sentence"},{"id":"T46","span":{"begin":0,"end":25},"obj":"Sentence"},{"id":"T47","span":{"begin":26,"end":198},"obj":"Sentence"},{"id":"T48","span":{"begin":199,"end":382},"obj":"Sentence"},{"id":"T49","span":{"begin":383,"end":456},"obj":"Sentence"},{"id":"T50","span":{"begin":457,"end":558},"obj":"Sentence"},{"id":"T51","span":{"begin":559,"end":967},"obj":"Sentence"},{"id":"T52","span":{"begin":968,"end":1076},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

simple1

{"project":"simple1","denotations":[{"id":"T7510","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7509","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7508","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7507","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7506","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

BioNLP16_DUT

{"project":"BioNLP16_DUT","denotations":[{"id":"T7953","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7952","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7951","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7950","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7949","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

BioNLP16_Messiy

{"project":"BioNLP16_Messiy","denotations":[{"id":"T7706","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7705","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7704","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7703","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7702","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

DLUT931

{"project":"DLUT931","denotations":[{"id":"T7916","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7915","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7914","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7913","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7912","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

bionlp-st-ge-2016-test-ihmc

{"project":"bionlp-st-ge-2016-test-ihmc","denotations":[{"id":"T7948","span":{"begin":968,"end":1001},"obj":"Binding"},{"id":"T7947","span":{"begin":26,"end":67},"obj":"Entity"},{"id":"T7946","span":{"begin":753,"end":765},"obj":"Entity"},{"id":"T7945","span":{"begin":968,"end":986},"obj":"Entity"},{"id":"T7944","span":{"begin":383,"end":395},"obj":"Entity"},{"id":"T7943","span":{"begin":706,"end":736},"obj":"Entity"},{"id":"T7942","span":{"begin":10,"end":25},"obj":"Entity"},{"id":"T7941","span":{"begin":300,"end":315},"obj":"Protein"},{"id":"T7940","span":{"begin":472,"end":474},"obj":"Protein"},{"id":"T7939","span":{"begin":517,"end":528},"obj":"Protein"},{"id":"T7938","span":{"begin":719,"end":735},"obj":"Entity"},{"id":"T7937","span":{"begin":1006,"end":1018},"obj":"Entity"},{"id":"T7936","span":{"begin":589,"end":608},"obj":"Entity"},{"id":"T7935","span":{"begin":776,"end":787},"obj":"Entity"},{"id":"T7934","span":{"begin":199,"end":211},"obj":"Entity"},{"id":"T7933","span":{"begin":62,"end":66},"obj":"Entity"},{"id":"T7932","span":{"begin":415,"end":417},"obj":"Protein"},{"id":"T7931","span":{"begin":533,"end":538},"obj":"Protein"},{"id":"T7930","span":{"begin":796,"end":814},"obj":"Entity"},{"id":"T7929","span":{"begin":231,"end":257},"obj":"Entity"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

bionlp-st-ge-2016-spacy-parsed

bionlp-st-ge-2016-test-tees

{"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T7500","span":{"begin":1006,"end":1010},"obj":"Protein"},{"id":"T7499","span":{"begin":533,"end":538},"obj":"Protein"},{"id":"T7498","span":{"begin":517,"end":528},"obj":"Protein"},{"id":"T7497","span":{"begin":383,"end":387},"obj":"Protein"},{"id":"T7496","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7495","span":{"begin":199,"end":203},"obj":"Protein"},{"id":"T7494","span":{"begin":13,"end":17},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

test3

{"project":"test3","denotations":[{"id":"T7338","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7337","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7336","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7335","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7334","span":{"begin":13,"end":16},"obj":"Protein"},{"id":"T7333","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7332","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7331","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7330","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7329","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

testone

{"project":"testone","denotations":[{"id":"T7321","span":{"begin":1006,"end":1009},"obj":"Protein"},{"id":"T7320","span":{"begin":383,"end":386},"obj":"Protein"},{"id":"T7319","span":{"begin":269,"end":272},"obj":"Protein"},{"id":"T7318","span":{"begin":199,"end":202},"obj":"Protein"},{"id":"T7317","span":{"begin":13,"end":16},"obj":"Protein"}],"text":"Isolation of CD4+ T cells\nPeripheral blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). CD4+ T cells were purified from PBMC by positive selection using anti-CD4 mAb-coated magnetic beads (Miltenyi Biotec, Gladbach, Germany), according to the manufacturer's instructions. CD4+ T cells were isolated from ST samples, as previously described [27]. Briefly, fresh ST samples were fragmented and digested with collagenase and DNase for 1 hour at 37°C. After removing tissue debris, ST cell suspensions in culture medium (RPMI 1640 medium; Life Technologies, Gaithersburg, MD, USA) supplemented with 25 mM HEPES (2 mM L-glutamine, 2% nonessential amino acids, 100 IU/ml penicillin, and 100 mg/ml streptomycin; Life Technologies) with 10% heat-inactivated FCS (Life Technologies) were incubated at 37°C in six-well plates (Coster, Cambridge, MA, USA) for 45 min. Non-adherent cells were harvested and CD4+ T cells were purified by positive selection as already described."}

PMC:1064873 / 8204-9280 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:1064873 / 8204-9280 JSON TXT