DOI: 10.1084/jem.193.11.1303. Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood

Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood

10.1084/jem.193.11.1303

Crossref journal-article

Rockefeller University Press

The Journal of Experimental Medicine (291)

Abstract

It has been known for years that rodents harbor a unique population of CD4+CD25+ “professional” regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4+CD25+CD45RO+ T cells (mean 6% of CD4+ T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4+CD25+ T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte–associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4+CD25+ T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4+CD25+ T cells released IL-10, and in coculture experiments suppressed the activation and proliferation of CD4+ and CD8+ T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4+CD25+ T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer.

Bibliography

Dieckmann, D., Plottner, H., Berchtold, S., Berger, T., & Schuler, G. (2001). Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood. The Journal of Experimental Medicine, 193(11), 1303â1310.

Authors 5

Detlef Dieckmann (first)
Heidi Plottner (additional)
Susanne Berchtold (additional)
Thomas Berger (additional)
Gerold Schuler (additional)

References 29 Referenced 852

10.1126/science.1900951 / Science. / Peripheral selection of the T cell repertoire by Rocha (1991)
10.1038/333742a0 / Nature. / Tolerance in T-cell-receptor transgenic mice involves deletion of nonmature CD4+CD8+ thymocytes by Kisielow (1988)
10.1126/science.2113314 / Science. / A cell culture model for T lymphocyte clonal anergy by Schwartz (1990)
10.1111/j.1600-065X.1993.tb01514.x / Immunol. Rev. / Self-ignorance in the peripheral T-cell pool by Miller (1993)
10.4049/jimmunol.155.3.1151 / J. Immunol. / Immunologic self-tolerance by activated T cells expressing IL-2 receptor α chains (CD25)breakdown of a single mechanism of self-tolerance causes various autoimmune disease by Sakaguchi (1995)
10.1093/intimm/10.12.1969 / Int. Immunol. / Immunologic self-tolerance maintained by CD25+CD4+ naturally anergic and suppressive T cellsinduction of autoimmune disease by breaking their anergic/suppressive state by Takahashi (1998)
10.4049/jimmunol.162.9.5317 / J. Immunol. / Thymus and autoimmunityproduction of CD4+CD25+ naturally anergic and suppressive T cells as a key function of the thymus in maintaining immunologic self-tolerance by Itoh (1999)
10.4049/jimmunol.163.10.5211 / J. Immunol. / Induction of tumor immunity by removing CD25+CD4+ T cellsa common basis between tumor immunity and autoimmunity by Shimizu (1999)
10.4049/jimmunol.164.1.183 / J. Immunol. / Suppressor effector function of CD4+CD25+ immunoregulatory T cells is antigen nonspecific by Thornton (2000)
10.1146/annurev.immunol.18.1.423 / Annu. Rev. Immunol. / Regulatory T cells in autoimmunity by Shevach (2000)
10.1084/jem.188.2.287 / J. Exp. Med. / CD4+CD25+ immunoregulatory T cells suppress polyclonal T cell activation in vitro by inhibiting interleukin 2 production by Thornton (1998)
10.1084/jem.192.2.295 / J. Exp. Med. / Cytotoxic T lymphocyte–associated antigen 4 plays an essential role in the function of CD25+CD4+ regulatory cells that control intestinal inflammation by Read (2000)
10.1016/S1074-7613(00)80195-8 / Immunity. / B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes by Salomon (2000)
10.1084/jem.192.2.303 / J. Exp. Med / Immunologic self-tolerance is maintained by CD25+CD4+ regulatory T cells constitutively expressing cytotoxic T lymphocyte–associated antigen 4 by Takahashi (2000)
10.1093/intimm/3.12.1349 / Int. Immunol. / A novel subpopulation of CD45RA+CD4+ T cells expressing IL-2 receptor α-chain (CD25) and having a functionally transitional nature into memory cells by Kanegane (1991)
{'key': '2023072509294354200_R16', 'first-page': '15', 'article-title': 'Preferential expression of IL-2 receptor subunits on memory population within CD4+ and CD8+ T cells', 'volume': '72', 'author': 'Taga', 'year': '1990', 'journal-title': 'Immunol.'} / Immunol. / Preferential expression of IL-2 receptor subunits on memory population within CD4+ and CD8+ T cells by Taga (1990)
10.1016/0090-1229(90)90012-F / Clin. Immunol. Immunopathol. / Restricted expression of p55 interleukin 2 receptor (CD25) on normal T cells by Jackson (1990)
10.1016/0022-1759(96)00078-6 / J. Immunol. Methods. / Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability by Romani (1996)
10.1016/S0022-1759(98)00208-7 / J. Immunol. Methods. / Generation of large numbers of fully mature and stable dendritic cells from leukapheresis products for clinical application by Thurner (1999)
10.1038/32588 / Nature. / Dendritic cells and the control of immunity by Bancherau (1998)
10.1038/39614 / Nature. / A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevent colitis by Groux (1997)
10.1172/JCI118779 / J. Clin. Invest. / Induction of circulating myelin basic protein and proteolipid protein-specific transforming growth factor-β1-secreting Th3 T cells by oral administration of myelin in multiple sclerosis patients by Fukaura (1996)
10.1016/S1074-7613(00)80366-0 / Immunity. / The emerging role of CTLA-4 as an immune attenuator by Thompson (1997)
10.1111/j.1600-065X.1996.tb00919.x / Immunol. Rev. / The role of CTLA-4 in the regulation and initiation of T-cell responses by Chambers (1996)
10.1084/jem.192.9.1213 / J. Exp. Med. / Induction of interleukin 10–producing, nonproliferating CD4+ T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells by Jonuleit (2000)
10.1093/intimm/10.4.371 / Int. Immunol. / Regulatory CD4 T cellsexpression of IL-2R α chain, resistance to clonal deletion and IL-2 dependency by Papiernik (1998)
10.1084/jem.189.5.877 / J. Exp. Med. / Peripheral autoantigen induces regulatory T cells that prevent autoimmunity by Seddon (1999)
10.1084/jem.191.3.411 / J. Exp. Med. / The induction of tolerance by dendritic cells that have captured apoptotic cells by Steinman (2000)
10.1084/jem.193.2.F5 / J. Exp. Med. / Differentiation of T regulatory cells by immature dendritic cells by Roncarolo (2001)

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:48 p.m.)
Deposited	2 years, 1 month ago (July 25, 2023, 6:43 a.m.)
Indexed	2 weeks ago (Aug. 23, 2025, 12:59 a.m.)
Issued	24 years, 3 months ago (June 4, 2001)
Published	24 years, 3 months ago (June 4, 2001)
Published Online	24 years, 3 months ago (June 4, 2001)
Published Print	24 years, 3 months ago (June 4, 2001)

Funders 0

None

BibTeX

@article{Dieckmann_2001, title={Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood}, volume={193}, ISSN={1540-9538}, url={http://dx.doi.org/10.1084/jem.193.11.1303}, DOI={10.1084/jem.193.11.1303}, number={11}, journal={The Journal of Experimental Medicine}, publisher={Rockefeller University Press}, author={Dieckmann, Detlef and Plottner, Heidi and Berchtold, Susanne and Berger, Thomas and Schuler, Gerold}, year={2001}, month=jun, pages={1303–1310} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        23
      ]
    ],
    "date-time": "2025-08-23T04:59:01Z",
    "timestamp": 1755925141494
  },
  "reference-count": 29,
  "publisher": "Rockefeller University Press",
  "issue": "11",
  "content-domain": {
    "domain": [
      "rupress.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2001,
        6,
        4
      ]
    ]
  },
  "abstract": "<jats:p>It has been known for years that rodents harbor a unique population of CD4+CD25+ \u201cprofessional\u201d regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4+CD25+CD45RO+ T cells (mean 6% of CD4+ T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4+CD25+ T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte\u2013associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4+CD25+ T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4+CD25+ T cells released IL-10, and in coculture experiments suppressed the activation and proliferation of CD4+ and CD8+ T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4+CD25+ T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer.</jats:p>",
  "DOI": "10.1084/jem.193.11.1303",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T16:48:33Z",
    "timestamp": 1027702113000
  },
  "page": "1303-1310",
  "update-policy": "http://dx.doi.org/10.1084/jem.crossmarkpolicy",
  "source": "Crossref",
  "is-referenced-by-count": 852,
  "title": "Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood",
  "prefix": "10.1084",
  "volume": "193",
  "author": [
    {
      "given": "Detlef",
      "family": "Dieckmann",
      "sequence": "first",
      "affiliation": [
        {
          "name": "aDepartment of Dermatology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany"
        }
      ]
    },
    {
      "given": "Heidi",
      "family": "Plottner",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "aDepartment of Dermatology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany"
        }
      ]
    },
    {
      "given": "Susanne",
      "family": "Berchtold",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "aDepartment of Dermatology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany"
        }
      ]
    },
    {
      "given": "Thomas",
      "family": "Berger",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "aDepartment of Dermatology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany"
        }
      ]
    },
    {
      "given": "Gerold",
      "family": "Schuler",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "aDepartment of Dermatology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany"
        }
      ]
    }
  ],
  "member": "291",
  "published-online": {
    "date-parts": [
      [
        2001,
        6,
        4
      ]
    ]
  },
  "reference": [
    {
      "key": "2023072509294354200_R1",
      "doi-asserted-by": "crossref",
      "first-page": "1225",
      "DOI": "10.1126/science.1900951",
      "article-title": "Peripheral selection of the T cell repertoire",
      "volume": "251",
      "author": "Rocha",
      "year": "1991",
      "journal-title": "Science."
    },
    {
      "key": "2023072509294354200_R2",
      "doi-asserted-by": "crossref",
      "first-page": "742",
      "DOI": "10.1038/333742a0",
      "article-title": "Tolerance in T-cell-receptor transgenic mice involves deletion of nonmature CD4+CD8+ thymocytes",
      "volume": "333",
      "author": "Kisielow",
      "year": "1988",
      "journal-title": "Nature."
    },
    {
      "key": "2023072509294354200_R3",
      "doi-asserted-by": "crossref",
      "first-page": "1349",
      "DOI": "10.1126/science.2113314",
      "article-title": "A cell culture model for T lymphocyte clonal anergy",
      "volume": "248",
      "author": "Schwartz",
      "year": "1990",
      "journal-title": "Science."
    },
    {
      "key": "2023072509294354200_R4",
      "doi-asserted-by": "crossref",
      "first-page": "131",
      "DOI": "10.1111/j.1600-065X.1993.tb01514.x",
      "article-title": "Self-ignorance in the peripheral T-cell pool",
      "volume": "133",
      "author": "Miller",
      "year": "1993",
      "journal-title": "Immunol. Rev."
    },
    {
      "key": "2023072509294354200_R5",
      "doi-asserted-by": "crossref",
      "first-page": "1151",
      "DOI": "10.4049/jimmunol.155.3.1151",
      "article-title": "Immunologic self-tolerance by activated T cells expressing IL-2 receptor \u03b1 chains (CD25)breakdown of a single mechanism of self-tolerance causes various autoimmune disease",
      "volume": "155",
      "author": "Sakaguchi",
      "year": "1995",
      "journal-title": "J. Immunol."
    },
    {
      "key": "2023072509294354200_R6",
      "doi-asserted-by": "crossref",
      "first-page": "1969",
      "DOI": "10.1093/intimm/10.12.1969",
      "article-title": "Immunologic self-tolerance maintained by CD25+CD4+ naturally anergic and suppressive T cellsinduction of autoimmune disease by breaking their anergic/suppressive state",
      "volume": "10",
      "author": "Takahashi",
      "year": "1998",
      "journal-title": "Int. Immunol."
    },
    {
      "key": "2023072509294354200_R7",
      "doi-asserted-by": "crossref",
      "first-page": "5317",
      "DOI": "10.4049/jimmunol.162.9.5317",
      "article-title": "Thymus and autoimmunityproduction of CD4+CD25+ naturally anergic and suppressive T cells as a key function of the thymus in maintaining immunologic self-tolerance",
      "volume": "162",
      "author": "Itoh",
      "year": "1999",
      "journal-title": "J. Immunol."
    },
    {
      "key": "2023072509294354200_R8",
      "doi-asserted-by": "crossref",
      "first-page": "5211",
      "DOI": "10.4049/jimmunol.163.10.5211",
      "article-title": "Induction of tumor immunity by removing CD25+CD4+ T cellsa common basis between tumor immunity and autoimmunity",
      "volume": "163",
      "author": "Shimizu",
      "year": "1999",
      "journal-title": "J. Immunol."
    },
    {
      "key": "2023072509294354200_R9",
      "doi-asserted-by": "crossref",
      "first-page": "183",
      "DOI": "10.4049/jimmunol.164.1.183",
      "article-title": "Suppressor effector function of CD4+CD25+ immunoregulatory T cells is antigen nonspecific",
      "volume": "164",
      "author": "Thornton",
      "year": "2000",
      "journal-title": "J. Immunol."
    },
    {
      "key": "2023072509294354200_R10",
      "doi-asserted-by": "crossref",
      "first-page": "423",
      "DOI": "10.1146/annurev.immunol.18.1.423",
      "article-title": "Regulatory T cells in autoimmunity",
      "volume": "18",
      "author": "Shevach",
      "year": "2000",
      "journal-title": "Annu. Rev. Immunol."
    },
    {
      "key": "2023072509294354200_R11",
      "doi-asserted-by": "crossref",
      "first-page": "287",
      "DOI": "10.1084/jem.188.2.287",
      "article-title": "CD4+CD25+ immunoregulatory T cells suppress polyclonal T cell activation in vitro by inhibiting interleukin 2 production",
      "volume": "188",
      "author": "Thornton",
      "year": "1998",
      "journal-title": "J. Exp. Med."
    },
    {
      "key": "2023072509294354200_R12",
      "doi-asserted-by": "crossref",
      "first-page": "295",
      "DOI": "10.1084/jem.192.2.295",
      "article-title": "Cytotoxic T lymphocyte\u2013associated antigen 4 plays an essential role in the function of CD25+CD4+ regulatory cells that control intestinal inflammation",
      "volume": "192",
      "author": "Read",
      "year": "2000",
      "journal-title": "J. Exp. Med."
    },
    {
      "key": "2023072509294354200_R13",
      "doi-asserted-by": "crossref",
      "first-page": "431",
      "DOI": "10.1016/S1074-7613(00)80195-8",
      "article-title": "B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes",
      "volume": "12",
      "author": "Salomon",
      "year": "2000",
      "journal-title": "Immunity."
    },
    {
      "key": "2023072509294354200_R14",
      "doi-asserted-by": "crossref",
      "first-page": "303",
      "DOI": "10.1084/jem.192.2.303",
      "article-title": "Immunologic self-tolerance is maintained by CD25+CD4+ regulatory T cells constitutively expressing cytotoxic T lymphocyte\u2013associated antigen 4",
      "volume": "192",
      "author": "Takahashi",
      "year": "2000",
      "journal-title": "J. Exp. Med"
    },
    {
      "key": "2023072509294354200_R15",
      "doi-asserted-by": "crossref",
      "first-page": "1349",
      "DOI": "10.1093/intimm/3.12.1349",
      "article-title": "A novel subpopulation of CD45RA+CD4+ T cells expressing IL-2 receptor \u03b1-chain (CD25) and having a functionally transitional nature into memory cells",
      "volume": "3",
      "author": "Kanegane",
      "year": "1991",
      "journal-title": "Int. Immunol."
    },
    {
      "key": "2023072509294354200_R16",
      "first-page": "15",
      "article-title": "Preferential expression of IL-2 receptor subunits on memory population within CD4+ and CD8+ T cells",
      "volume": "72",
      "author": "Taga",
      "year": "1990",
      "journal-title": "Immunol."
    },
    {
      "key": "2023072509294354200_R17",
      "doi-asserted-by": "crossref",
      "first-page": "126",
      "DOI": "10.1016/0090-1229(90)90012-F",
      "article-title": "Restricted expression of p55 interleukin 2 receptor (CD25) on normal T cells",
      "volume": "54",
      "author": "Jackson",
      "year": "1990",
      "journal-title": "Clin. Immunol. Immunopathol."
    },
    {
      "key": "2023072509294354200_R18",
      "doi-asserted-by": "crossref",
      "first-page": "137",
      "DOI": "10.1016/0022-1759(96)00078-6",
      "article-title": "Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability",
      "volume": "196",
      "author": "Romani",
      "year": "1996",
      "journal-title": "J. Immunol. Methods."
    },
    {
      "key": "2023072509294354200_R19",
      "doi-asserted-by": "crossref",
      "first-page": "1",
      "DOI": "10.1016/S0022-1759(98)00208-7",
      "article-title": "Generation of large numbers of fully mature and stable dendritic cells from leukapheresis products for clinical application",
      "volume": "223",
      "author": "Thurner",
      "year": "1999",
      "journal-title": "J. Immunol. Methods."
    },
    {
      "key": "2023072509294354200_R20",
      "doi-asserted-by": "crossref",
      "first-page": "245",
      "DOI": "10.1038/32588",
      "article-title": "Dendritic cells and the control of immunity",
      "volume": "392",
      "author": "Bancherau",
      "year": "1998",
      "journal-title": "Nature."
    },
    {
      "key": "2023072509294354200_R21",
      "doi-asserted-by": "crossref",
      "first-page": "737",
      "DOI": "10.1038/39614",
      "article-title": "A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevent colitis",
      "volume": "389",
      "author": "Groux",
      "year": "1997",
      "journal-title": "Nature."
    },
    {
      "key": "2023072509294354200_R22",
      "doi-asserted-by": "crossref",
      "first-page": "70",
      "DOI": "10.1172/JCI118779",
      "article-title": "Induction of circulating myelin basic protein and proteolipid protein-specific transforming growth factor-\u03b21-secreting Th3 T cells by oral administration of myelin in multiple sclerosis patients",
      "volume": "98",
      "author": "Fukaura",
      "year": "1996",
      "journal-title": "J. Clin. Invest."
    },
    {
      "key": "2023072509294354200_R23",
      "doi-asserted-by": "crossref",
      "first-page": "445",
      "DOI": "10.1016/S1074-7613(00)80366-0",
      "article-title": "The emerging role of CTLA-4 as an immune attenuator",
      "volume": "7",
      "author": "Thompson",
      "year": "1997",
      "journal-title": "Immunity."
    },
    {
      "key": "2023072509294354200_R24",
      "doi-asserted-by": "crossref",
      "first-page": "27",
      "DOI": "10.1111/j.1600-065X.1996.tb00919.x",
      "article-title": "The role of CTLA-4 in the regulation and initiation of T-cell responses",
      "volume": "153",
      "author": "Chambers",
      "year": "1996",
      "journal-title": "Immunol. Rev."
    },
    {
      "key": "2023072509294354200_R25",
      "doi-asserted-by": "crossref",
      "first-page": "1213",
      "DOI": "10.1084/jem.192.9.1213",
      "article-title": "Induction of interleukin 10\u2013producing, nonproliferating CD4+ T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells",
      "volume": "192",
      "author": "Jonuleit",
      "year": "2000",
      "journal-title": "J. Exp. Med."
    },
    {
      "key": "2023072509294354200_R26",
      "doi-asserted-by": "crossref",
      "first-page": "5317",
      "DOI": "10.1093/intimm/10.4.371",
      "article-title": "Regulatory CD4 T cellsexpression of IL-2R \u03b1 chain, resistance to clonal deletion and IL-2 dependency",
      "volume": "10",
      "author": "Papiernik",
      "year": "1998",
      "journal-title": "Int. Immunol."
    },
    {
      "key": "2023072509294354200_R27",
      "doi-asserted-by": "crossref",
      "first-page": "877",
      "DOI": "10.1084/jem.189.5.877",
      "article-title": "Peripheral autoantigen induces regulatory T cells that prevent autoimmunity",
      "volume": "189",
      "author": "Seddon",
      "year": "1999",
      "journal-title": "J. Exp. Med."
    },
    {
      "key": "2023072509294354200_R28",
      "doi-asserted-by": "crossref",
      "first-page": "411",
      "DOI": "10.1084/jem.191.3.411",
      "article-title": "The induction of tolerance by dendritic cells that have captured apoptotic cells",
      "volume": "191",
      "author": "Steinman",
      "year": "2000",
      "journal-title": "J. Exp. Med."
    },
    {
      "key": "2023072509294354200_R29",
      "doi-asserted-by": "crossref",
      "first-page": "F5",
      "DOI": "10.1084/jem.193.2.F5",
      "article-title": "Differentiation of T regulatory cells by immature dendritic cells",
      "volume": "193",
      "author": "Roncarolo",
      "year": "2001",
      "journal-title": "J. Exp. Med."
    }
  ],
  "container-title": "The Journal of Experimental Medicine",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://rupress.org/jem/article-pdf/193/11/1303/1700201/010432.pdf",
      "content-type": "application/pdf",
      "content-version": "vor",
      "intended-application": "syndication"
    },
    {
      "URL": "https://rupress.org/jem/article-pdf/193/11/1303/1700201/010432.pdf",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        7,
        25
      ]
    ],
    "date-time": "2023-07-25T10:43:58Z",
    "timestamp": 1690281838000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://rupress.org/jem/article/193/11/1303/25830/Ex-Vivo-Isolation-and-Characterization-of-Cd4-Cd25"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2001,
        6,
        4
      ]
    ]
  },
  "references-count": 29,
  "journal-issue": {
    "issue": "11",
    "published-print": {
      "date-parts": [
        [
          2001,
          6,
          4
        ]
      ]
    }
  },
  "URL": "http://dx.doi.org/10.1084/jem.193.11.1303",
  "relation": {},
  "ISSN": [
    "0022-1007",
    "1540-9538"
  ],
  "subject": [],
  "published": {
    "date-parts": [
      [
        2001,
        6,
        4
      ]
    ]
  }
}