PMC:1064873 / 8230-9280 JSONTXT

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    LappsTest

    {"project":"LappsTest","denotations":[{"id":"T7328","span":{"begin":980,"end":984},"obj":"Protein"},{"id":"T7327","span":{"begin":507,"end":512},"obj":"Protein"},{"id":"T7326","span":{"begin":491,"end":502},"obj":"Protein"},{"id":"T7325","span":{"begin":357,"end":361},"obj":"Protein"},{"id":"T7324","span":{"begin":238,"end":246},"obj":"Protein"},{"id":"T7323","span":{"begin":173,"end":177},"obj":"Protein"}],"text":"Peripheral 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":426,"end":428},"obj":"9182920"}],"text":"Peripheral 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

    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blood mononuclear cells (PBMC) were prepared from heparinized blood samples by centrifugation over Ficoll-Hypaque density gradients (Pharmacia, Uppsala, Sweden). 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    pmc-enju-pas

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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-proteins

    {"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T7482","span":{"begin":980,"end":983},"obj":"Protein"},{"id":"T7481","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7480","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7479","span":{"begin":173,"end":176},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"Peripheral 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":980,"end":983},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7492","span":{"begin":357,"end":360},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7491","span":{"begin":243,"end":246},"obj":"http://www.uniprot.org/uniprot/P01730"},{"id":"T7490","span":{"begin":173,"end":176},"obj":"http://www.uniprot.org/uniprot/P01730"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Peripheral 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":548,"end":554},"obj":"http://purl.obolibrary.org/obo/UBERON_0000479"},{"id":"T7340","span":{"begin":73,"end":78},"obj":"http://purl.obolibrary.org/obo/UBERON_0000178"},{"id":"T7339","span":{"begin":11,"end":16},"obj":"http://purl.obolibrary.org/obo/UBERON_0000178"}],"text":"Peripheral 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":987,"end":992},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7487","span":{"begin":955,"end":960},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7486","span":{"begin":364,"end":369},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7485","span":{"begin":180,"end":185},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T7484","span":{"begin":29,"end":34},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Peripheral 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":942,"end":1050},"obj":"Sentence"},{"id":"T7471","span":{"begin":431,"end":941},"obj":"Sentence"},{"id":"T7470","span":{"begin":357,"end":430},"obj":"Sentence"},{"id":"T7469","span":{"begin":173,"end":356},"obj":"Sentence"},{"id":"T7468","span":{"begin":0,"end":172},"obj":"Sentence"},{"id":"T47","span":{"begin":0,"end":172},"obj":"Sentence"},{"id":"T48","span":{"begin":173,"end":356},"obj":"Sentence"},{"id":"T49","span":{"begin":357,"end":430},"obj":"Sentence"},{"id":"T50","span":{"begin":431,"end":532},"obj":"Sentence"},{"id":"T51","span":{"begin":533,"end":941},"obj":"Sentence"},{"id":"T52","span":{"begin":942,"end":1050},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7509","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7508","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7507","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7952","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7951","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7950","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7705","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7704","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7703","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7915","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7914","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7913","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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":942,"end":975},"obj":"Binding"},{"id":"T7947","span":{"begin":0,"end":41},"obj":"Entity"},{"id":"T7946","span":{"begin":727,"end":739},"obj":"Entity"},{"id":"T7945","span":{"begin":942,"end":960},"obj":"Entity"},{"id":"T7944","span":{"begin":357,"end":369},"obj":"Entity"},{"id":"T7943","span":{"begin":680,"end":710},"obj":"Entity"},{"id":"T7941","span":{"begin":274,"end":289},"obj":"Protein"},{"id":"T7940","span":{"begin":446,"end":448},"obj":"Protein"},{"id":"T7939","span":{"begin":491,"end":502},"obj":"Protein"},{"id":"T7938","span":{"begin":693,"end":709},"obj":"Entity"},{"id":"T7937","span":{"begin":980,"end":992},"obj":"Entity"},{"id":"T7936","span":{"begin":563,"end":582},"obj":"Entity"},{"id":"T7935","span":{"begin":750,"end":761},"obj":"Entity"},{"id":"T7934","span":{"begin":173,"end":185},"obj":"Entity"},{"id":"T7933","span":{"begin":36,"end":40},"obj":"Entity"},{"id":"T7932","span":{"begin":389,"end":391},"obj":"Protein"},{"id":"T7931","span":{"begin":507,"end":512},"obj":"Protein"},{"id":"T7930","span":{"begin":770,"end":788},"obj":"Entity"},{"id":"T7929","span":{"begin":205,"end":231},"obj":"Entity"}],"text":"Peripheral 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

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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-tees

    {"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T7500","span":{"begin":980,"end":984},"obj":"Protein"},{"id":"T7499","span":{"begin":507,"end":512},"obj":"Protein"},{"id":"T7498","span":{"begin":491,"end":502},"obj":"Protein"},{"id":"T7497","span":{"begin":357,"end":361},"obj":"Protein"},{"id":"T7496","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7495","span":{"begin":173,"end":177},"obj":"Protein"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7337","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7336","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7335","span":{"begin":173,"end":176},"obj":"Protein"},{"id":"T7333","span":{"begin":980,"end":983},"obj":"Protein"},{"id":"T7332","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7331","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7330","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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":980,"end":983},"obj":"Protein"},{"id":"T7320","span":{"begin":357,"end":360},"obj":"Protein"},{"id":"T7319","span":{"begin":243,"end":246},"obj":"Protein"},{"id":"T7318","span":{"begin":173,"end":176},"obj":"Protein"}],"text":"Peripheral 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."}