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The humoral immune response to head and neck cancer antigens as defined by the serological analysis of tumor antigens by recombinant cDNA expression cloning

Hilary A. Vaughan, Fiona St Clair, Matthew J. Scanlan, Yao-Tsen Chen, Eugene Maraskovsky, Andrew Sizeland, Lloyd J. Old and Jonathan Cebon
Hilary A. Vaughan
1Cancer Vaccine Laboratory, Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, 3084, Australia
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Fiona St Clair
1Cancer Vaccine Laboratory, Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, 3084, Australia
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Matthew J. Scanlan
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA
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Yao-Tsen Chen
3Dept. of Pathology, Weill Medical College of Cornell University, New York, USA
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Eugene Maraskovsky
1Cancer Vaccine Laboratory, Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, 3084, Australia
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Andrew Sizeland
4Peter MacCallum Cancer Institute, Melbourne, Australia
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Lloyd J. Old
2Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA
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Jonathan Cebon
1Cancer Vaccine Laboratory, Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, 3084, Australia
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DOI:  Published January 2004
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  • Figure 1
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    Figure 1

    Northern blot analysis of HNSCC SEREX clones. The expression of clones AU-HN-4 (A), AU-HN-18 (B), AU-HN-25 (C), and AU-HN-6 (D) was assayed in the following normal human tissues: (1) spleen, (2) thymus, (3) prostate, (4) testis, (5) ovary, (6) small intestine, (7) colon, and (8) lymphocytes.

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    Figure 2

    AU-HN-15 expression in HNSCC tumors. The expression of AU-HN-15 (A) and beta-actin (B) was determined by RT-PCR in the following samples: (1-8) HNSCC tumor tissue; (9-12) HNSCC cell lines: SCC-4 (9), HTB-43 (10), SCC-25 (11) and CCL-30 (12); (13-16) tissue adjacent to HNSCC tumor; (17-18) AU-HN-15 plasmid.

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    Figure 3

    Quantification of clone AU-HN-15 mRNA in normal and tumor tissues by qRT-PCR. Messenger RNA copy number is expressed as the number of AU-HN-15 molecules in 100 ng of total RNA per 105 molecules of beta-glucoronidase. (A) Normal tissues: (1) brain, (2) testis-1, (3) colon, (4) liver, (5) testis-2, (6) kidney-1, (7) lung, (8) trachea, (9) small intestine, (10) stomach, (11) thymus, (12) heart, (13), spleen, (14) bone marrow, (15) kidney-2. (B) HNSCC tumor tissues (1-11) and HNSCC cell lines (12-15): SCC-25 (12), HTB-4B (13), SCC-4 (14), CCL-30 (15). (C) Paired samples: HNSCC (1, 3, 5) and adjacent mucosae (2, 4, 6) from three primary tumors; HNSCC metastatic to lymph node (7) and paired normal tissue (8).

Tables

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  • Table 1

    HNSCC clones corresponding to known genes.

    Functional Group Clone ID No. of Times Isolated Identity Unigene Cluster Previously Identified by SEREX
    DNA binding/ Transcription AU-HN-2 1 TREX1, 3' repair exonuclease Hs.278408 no
    AU-HN-4 2 BRAP (BRCA1 associated): Nuclear localization protein Hs.122764 yes
    AU-HN-5 1 Trim 26 zinc finger domains Hs.1287 no
    AU-HN-10 1 GTF21 transcription factor. Murine homolog TF11-1 Hs.278589 no
    AU-HN-11 6 NSEP1 (YB-1) transcription factor Hs.74497 yes
    AU-HN-16 1 MAZ transcription factor associated with c-myc Hs.7647 yes
    AU-HN-17 1 SON (DBP-5; KIAA1019; NREBP DNA binding protein) Hs.92909 yes
    AU-HN-27 1 NACA nascent polypeptide-associated complex Hs.32916 yes
    AU-HN-37 1 NUBP2 nucleotide binding protein Hs.256549 no
    RNA processing/ Translation AU-HN-3 1 EEF2 Translation elongation factor 2 Hs.75309 no
    AU-HN-13 2 GU2 Putative RNA helicase Hs.7392 no
    AU-HN-30 1 RPLI3A ribosomal protein Hs.119122 no
    AU-HN-31 1 SFRS21P (CASP11; SIP1; SRRP1290 splicing factor) Hs.51957 no
    AU-HN-33 1 RPS12 ribosomal protein Hs.339696 no
    AU-HN-34 1 MGC2835 RNA helicase Hs.70582 no
    Regulatory AU-HN-18 1 TMF1, TATA modulatory factor Hs.267632 yes
    AU-HN-23 1 PRC1 regulator of cytokinesis Hs.344037 no
    Structural AU-HN-22 2 KRT14 keratin 14 Hs.355214 no
    AU-HN-28 1 Viniculin Hs.75350 yes
    AU-HN-32 1 H2AFY histone family member Hs.75258 no
    Apoptosis repressor AU-HN-14 2 SLK (KIAA02304) Ste related kinase Hs.105751 no
    AU-HN-20 2 NOL3 (ARC) nuclear protein 3, apoptosis repressor Hs.278439 no
    Molecular chaperone AU-HN-24 1 DNAJA2 member of Hsp40 family Hs.21189 yes
    AU-HN-35 1 DNAJA1 member of HSP40 family Hs.94 yes
    Other AU-HN-1 1 LINE-1 retrotransposon No yes
    AU-HN-6 1 MOG (HSPC 165) Homolog of yeast protein Hs.13605 no
    AU-HN-12 4 LIMS1 (PINCH): LIM and senescent antigen-like domain Hs.112378, Hs.193700 yes
    AU-HN-21 2 COPB2 coatomer protein complex subunit protein Hs.75724 no
    AU-HN-25 1 FLJ22548 hypothetical protein Hs.103267 no
    AU-HN-26 2 C21orf97; FLJ21324; MGC15873 Hs.4746 yes
    AU-HN-29 3 SSNA1 Sjogrens syndrome nuclear autoantigen 1 Hs.18528 no
  • Table 2

    Correspondence of HNSCC clones with other SEREX clones.

    Clone ID SEREX Clones Tumor Type
    AU-HN-1 NGO-Lu-8 lung
    AU-HN-4 P2B-25f, NGO-Br-5, NY-REN-63, MO-BR-100, RMNY-BR-52, MO-OVA-76, NY-CO-6, HOM-TS-GL1-16, TM-92, TE2-7 breast, renal, ovary, colon, normal testis
    AU-HN-11 HOM-TSOv2.35, HOM-Ts-PMR2-12 normal testis
    AU-HN-12 NGO-St-79, MO-REN-80, NY-REN-48, NGO-Br-47, NY-ESO-10, UL-AML-4, NGO-Co-8 stomach, renal, breast, esophagus, AML, colon
    AU-HN-14 MO-BC-1128, se20-9, NGO-Lu-33 lung
    AU-HN-16 MO-CO-135, MO-CO-2 colon
    AU-HN-17 NGO-Pr-44 prostate
    AU-HN-18 NGO-St-29, LONY-BR-81, MO-CO-103 stomach, breast, colon
    AU-HN-24 NY-REN-14 renal
    AU-HN-26 NY-CO-18 colon
    AU-HN-27 NY-CO-2, NW-FW 22, NGO-Lu-27 colon, lung
    AU-HN-28 NY-BR-101, NGO-Br-52 breast
    AU-HN-35 NGO-Lu-17, 6-32, 6-25.1, 6-8.1, 6.26.1, 6.27 lung, liver
  • Table 3

    HNSCC SEREX clones corresponding to novel or not fully characterized genes.

    Function Clone No. Identity Previously Identified SEREXa Clones Tumor Type Unigene Cluster
    Regulatory/ DNA binding AU-HN-9 unknown no   Hs.95262
    AU-HN-15 KIAA0530, zinc finger domain HOM-TSsemA-56, NY-CO-6, MOBr-17 colon, breast, normal testis Hs.173081
    TNF-inducible AU-HN-36 rat stannin no   Hs.76691
    Unknown AU-HN-7 hypothetical protein DKFZp4340032 no   Hs.114173
    AU-HN-8 human FLJ23089 no   Hs.130554
    AU-HN-19 PC326 HOM-TSsem4.95, P2A-89, HOM-TSMa4.9, MOVA95, HOM-HCC-27.8, HOM-Br3.40, Mz19-F-45 prostate, liver,melanoma, lung,ovary, normal testis Hs.279882

    aSEREX database (36).

    • Table 4

      Novel antigens identified in HNSCC by SEREX: summary of Northern blot analyses.

      Tissue Expressiona of Clone AU-HN-
      2 4 6 9 14 15 18 20 23 25 34
      Spleen ++ - - + + - - + nt - nt
      Thymus + - - + + - - + nt (+) nt
      Prostate ++ - - + + - - + nt + nt
      Testis + ++ +++ + + - ++ +++ nt +++ nt
      Lymphocytes ++ - - + + - - + nt (+) nt
      Ovary + - - + + - - + nt + nt
      Small intestine + - - + + - - + nt - nt
      Colon + - - + + - - + - + -
      Brain nt nt nt nt nt - nt nt - nt +
      Heart nt nt nt nt nt - nt nt - nt +
      Kidney nt nt nt nt nt - nt nt ++ nt +
      Liver nt nt nt nt nt - nt nt ++ nt +
      Lung nt nt nt nt nt - nt nt - nt -

      aExpression: +++, high expression; ++, medium expression; +, low expression; (+), very weak; nt, not tested.

      • Table 5

        Serological screening of the HNSCC screen 5 clones with the individual HNSCC sera making up the pool used for the screening.

        Clone No. Identity Serum
        BT RM DG
        AU-HN-12 PINCH + + +
        AU-HN-22 keratin - - +
        AU-HN-23 human Hsp DNAj (+)a - -
        AU-HN-25 FLJ22548 - - +
        AU-HN-26a C21orf97, FDJ21324 (+) - -
        AU-HN-26b + + -
        AU-HN-27 NACN, transcription activator - + +
        AU-HN-28 vinculin - + +
        AU-HN-29a Sjogren's nuclear antigen NA14 + - +
        AU-HN-29b (+) - -
        AU-HN-29c + + +
        AU-HN-30 L13a ribosomal protein (+) - -
        AU-HN-31 SFRS21 splicing factor - + -
        AU-HN-32 H2A.1 histone + - +
        AU-HN-33 S12 ribosomal protein - + -
        AU-HN-34 RNA helicase related (+) - -
        AU-HN-35 human Hsp DNAj (+) - -
        AU-HN-36 rat stannin related + + -
        AU-HN-97 laminin receptor + + +

        a(+) HNSCC clones positive only with BT serum (adenocarcinoma).

        • Table 6

          Description of SEREX screenings.

          Screen No. Tissue Serum (dilution) No. of pfu Screened No. of Clones
          1 JKa tumor JKa (1:100) 5 x 105 3
          2 normal testis JKa (1:100) 1 x 106 9
          4 SCC-25b RLc (1:100) 1 x 106 19
          5 RMd tumor pool: RMd (1:100), DGe (1:150) and BTf (1:200) 9 x 105 19

          aJK: Right neck dissection, LN with metastatic moderately differentiated keratinizing SCC. bSCC-25: Cell line established from SCC of the tongue. cRL: Neck dissection, metastatic moderately differentiated SCC from primary tongue lesion; large tumor burden. dRM: Left cheek tumor, ulcerating moderately differentiated keratinizing SCC. eDG: HNSCC Stage 4; large tumor burden. fBT: Neck dissection, metastatic adenocarcinoma.

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          Cancer Immunity Archive: 4 (1)
          January 2004
          Volume 4, Issue 1
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          The humoral immune response to head and neck cancer antigens as defined by the serological analysis of tumor antigens by recombinant cDNA expression cloning
          Hilary A. Vaughan, Fiona St Clair, Matthew J. Scanlan, Yao-Tsen Chen, Eugene Maraskovsky, Andrew Sizeland, Lloyd J. Old and Jonathan Cebon
          Cancer Immun January 1 2004 (4) (1) 5;

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          The humoral immune response to head and neck cancer antigens as defined by the serological analysis of tumor antigens by recombinant cDNA expression cloning
          Hilary A. Vaughan, Fiona St Clair, Matthew J. Scanlan, Yao-Tsen Chen, Eugene Maraskovsky, Andrew Sizeland, Lloyd J. Old and Jonathan Cebon
          Cancer Immun January 1 2004 (4) (1) 5;
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