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HLA-DP4 expression and immunity to NY-ESO-1: correlation and characterization of cytotoxic CD4+ CD25- CD8- T cell clones

Eduardo Huarte, Julia Karbach, Sacha Gnjatic, Armin Bender, Dirk Jäger, Michael Arand, Djordje Atanackovic, Jonathan Skipper, Gerd Ritter, Yao-Tseng Chen, Lloyd J. Old, Alexander Knuth and Elke Jäger
Eduardo Huarte
1II. Medizinische Klinik, Hämatologie-Onkologie, Krankenhaus Nordwest, Frankfurt, Germany
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Julia Karbach
1II. Medizinische Klinik, Hämatologie-Onkologie, Krankenhaus Nordwest, Frankfurt, Germany
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Sacha Gnjatic
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Armin Bender
1II. Medizinische Klinik, Hämatologie-Onkologie, Krankenhaus Nordwest, Frankfurt, Germany
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Dirk Jäger
3Abteilung für Onkologie, Universitätsspital Zürich, Switzerland
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Michael Arand
4Institut für Pharmakologie und Toxikologie, Universität Würzburg, Germany
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Djordje Atanackovic
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Jonathan Skipper
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Gerd Ritter
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Yao-Tseng Chen
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Lloyd J. Old
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, USA
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Alexander Knuth
3Abteilung für Onkologie, Universitätsspital Zürich, Switzerland
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Elke Jäger
1II. Medizinische Klinik, Hämatologie-Onkologie, Krankenhaus Nordwest, Frankfurt, Germany
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DOI:  Published January 2004
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    Figure 1

    Induction of NY-ESO-1-specific CD4+ and CD8+ T cell responses after in vitro presensitization with NY-ESO-1 p157-170 or Ad2/ESO. CD4+ and CD8+ T cells were presensitized with (a) NY-ESO-1 p157-170 or (b) Ad2/ESO. Two patients, NW1454 and NW1691, showed nonspecific reactivity against the irrelevant Melan A peptide p29-37 (white bars) after Ad2/ESO stimulation (a and b). (c) CD8+ T cell responses were induced after presensitization with NY-ESO-1 p157-170 against the stimulating peptide (black bar), and against T2 cells pulsed with NY-ESO-1 p157-165 (checkered bars) and p157-167 (vertical stripes). No significant reactivity was observed against the irrelevant Melan A peptide p29-37 (white bars) or against T2 cells alone (horizontal stripes).

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    NY-ESO-1 p157-170-specific CD4+ T cell reactivity is restricted by HLA-DP4. The CD4+ T cell clone NW 1662-CD4-12 was tested against a panel of HLA-DP4-positive and HLA-DP4-negative allogeneic EBV-B cell lines pulsed with NY-ESO-1 p157-170 (black bars) in ELISPOT assays. NY-ESO-1 p157-170-specific reactivity was documented against peptide-pulsed HLA-DP4+ EBV-B cells only. The white bars represent the background reactivity against EBV-B cells alone.

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    Figure 3

    Reactivity of HLA-DP4-restricted NY-ESO-1-specific CD4+ T cells clones against different concentrations of NY-ESO-1 p157-170. Seven CD4+ T cell clones obtained from patients NW1454 and NW1662 were tested against different concentrations of NY-ESO-1 p157-170 in ELISPOT assays. Depending upon the interclonal avidity, different levels of peptide concentration are required for detectable reactivity with the respective CD4+ T cell clone.

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    Figure 4

    Recognition of naturally processed NY-ESO-1. (a) A panel of CD4+ T cell clones were tested in ELISPOT assays against autologous DCs pulsed with recombinant NY-ESO-1 protein (black bars) or irrelevant recombinant SSX protein (white bars), or infected with Ad2/ESO (horizontal stripes), v.v. ESO (checkered bars), or v.v. WT (gray bars). (b) CD4+ T cell clone NW 1662-CD4-2 was tested for specific recognition of autologous DCs pulsed with lysates prepared from NY-ESO-1-positive (NW-MEL-38 and SK-MEL-37) or NY-ESO-1-negative (NW-MEL-8) melanoma cell lines in ELISPOT assays.

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    Figure 5

    NY-ESO-1-specific cytotoxic reactivity of CD4+ T cell clones. (a) The CD4+ T cell clones NW 1454-CD4-68 (triangles) and NW 1662-CD4-2 (squares) were tested against the allogenic HLA-DP4+ EBV-B cell line NW1539-EBV-B, either pulsed with NY-ESO-1 p157-170 or not (closed and open symbols, respectively) in a 4-h chromium release assay at E/T ratios of 90, 30, 10, and 1:1. (b) The CD4+ T cell clone NW 1454-CD4-68 was tested against the HLA-DP4+ NY-ESO-1-positive melanoma cell lines NW-MEL-38, NW-MEL 634, and NW-MEL-450, against the NY-ESO-1-negative melanoma cell line NW-MEL-8, and against the HLA-DP4-negative melanoma cell line SK-MEL-29 in a 4-h chromium release assay.

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    NY-ESO-1 antibody and HLA-DP4 status of patients with NY-ESO-1-expressing cancers.

    HLA-DP4 status NY-ESO-1 Ab status
    Ab+ patients (n=50) Ab- patients (n=52)
    HLA-DP4+ 39 40
    HLA-DP4- 11 12
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Cancer Immunity Archive: 4 (1)
January 2004
Volume 4, Issue 1
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HLA-DP4 expression and immunity to NY-ESO-1: correlation and characterization of cytotoxic CD4+ CD25- CD8- T cell clones
Eduardo Huarte, Julia Karbach, Sacha Gnjatic, Armin Bender, Dirk Jäger, Michael Arand, Djordje Atanackovic, Jonathan Skipper, Gerd Ritter, Yao-Tseng Chen, Lloyd J. Old, Alexander Knuth and Elke Jäger
Cancer Immun January 1 2004 (4) (1) 15;

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HLA-DP4 expression and immunity to NY-ESO-1: correlation and characterization of cytotoxic CD4+ CD25- CD8- T cell clones
Eduardo Huarte, Julia Karbach, Sacha Gnjatic, Armin Bender, Dirk Jäger, Michael Arand, Djordje Atanackovic, Jonathan Skipper, Gerd Ritter, Yao-Tseng Chen, Lloyd J. Old, Alexander Knuth and Elke Jäger
Cancer Immun January 1 2004 (4) (1) 15;
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