cerevisiae) PMS1 NM_000534 231.3029

  retinoblastoma binding protein 8, transcript variant 1 RBBP8 NM_002894 check details 332.3025 473.1274 ribosomal protein, large, P0, transcript variant 1 RPLP0 NM_001002 179.1131 433.1217 RNA export 1 homolog (S.pombe), transcript variant 1 RAE1 NM_003610 342.1448   serine/threonine kinase 3(STE20 homolog, yeast) STK3 NM_006281 142.1617   SH3-domain GRB2-like 1 SH3GL1 NM_003025 107.1213 43.1615 synaptonemal complex protein SC65 SC65 NM_006455 289.1598   TAF7 RNA polymerase II, TATA box binding protein (TBP)-associated factor, 55 kDa TAF7 NM_005642 741.1790 1578.2310 talin 1 TLN1 NM_006289 91.7716 5712…8187 transforming growth factor, beta-induced, 68 kDa TGFBI NM_000358 48.2099 1371…2691 unc-45 homolog A (C.elegans), transcript variant 2 or 3 UNC45A NM_001039675 836.3625 1924.3471 † cDNA inserts of positive clones were successfully expressed into proteins followed by ELISA. The GST-fusion recombinant proteins were successfully produced using pGEX-4 T vectors in 10 of 31 antigens—centromere protein F, 350/400 ka (CENPF); macrophage migration inhibitory factor (MIF); myosin phosphatase-Rho interacting protein (M-RIP); retinoblastoma binding protein 8 (RBBP8); ribosomal protein, large, P0 (RPLP0); SH3GL1, TAF7 RNA polymerase II, TATA box binding protein-associated factor, 55 kDa (TAF7); talin 1 (TLN1); transforming growth factor beta-induced BYL719 molecular weight 68 kDa

(TGFBI), and unc-45 homolog A (UNC45A) (Figures 1 and 2). Figure 1 Serum antibody levels of glioma Branched chain aminotransferase SEREX antigens. cDNA inserts of identified clones were recombined in-frame into pGEX vectors that express recombinant GST fusion proteins. Using the fusion proteins as antigens, the

levels of antibodies were examined by the ELISA and shown by the ordinate, as (A) CENPF, (B) MIF, (C) M-RIP, (D) RBBP8, (E) RPLP0, (F) TAF7, (G) TLN1, (H) TGFBI, (I) UNC45A. The significance of differences among healthy donors, patients with low-grade glioma and with high-grade glioma was calculated using Kruskal Wallis H-test and Mann–Whitney U-test with Bonferroni correction. The box-and-whisker plots display the 10th, 25th, 50th, 75th and 90th percentiles. Figure 2 The increasing levels of antibodies to SH3GL1 in sera of the patients with low-grade glioma. Serum antibody level to SH3GL1 was examined by the ELISA as described in the legends of Figure 1. First screening test (A) and the individual validation test (B), revealed the significant higher levels of autologous antibody against SH3GL1 in low-grade glioma patients, than healthy donors (P = 0.045 and 0.0189). ELISA to detect serum antibodies Using a recombinant antigen protein, ELISA was performed on sera from 32 patients with high-grade glioma, 40 with low-grade glioma and 56 healthy volunteers, which were collected between 1998 and 2005 in Chiba University Hospital. The serum used for SEREX screening was excluded. The characteristics of the sera are shown in Table  2 (left).