Oncogene (2001) 20, 4107 ± 4114 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc ORIGINAL PAPERS PHF3-speci®c antibody responses in over 60% of patients with glioblastoma multiforme

A-K Struss1, BFM Romeike2, A Munnia1, W Nastainczyk3, W-I Steudel4,JKoÈ nig5, H Ohgaki6, W Feiden2, U Fischer1,7 and E Meese*,1,7

1Institut fuÈr Humangenetik, UniversitaÈt(-skliniken) des Saarlandes, 66421 Homburg/Saar, Germany; 2Abteilung fuÈr Neuropathologie, UniversitaÈt(-skliniken) des Saarlandes, 66421 Homburg/Saar, Germany; 3Institut fuÈr Medizinische Biochemie, UniversitaÈt(-skliniken) des Saarlandes, 66421 Homburg/Saar, Germany; 4Neurochirurgische Klinik, UniversitaÈt(-skliniken) des Saarlandes, 66421 Homburg/Saar, Germany; 5Institut fuÈr Bioinformatik; UniversitaÈt(-skliniken) des Saarlandes, 66421 Homburg/ Saar, Germany; 6International Agency for Research on Cancer, 150 Cours Albert Thomas, 69372 Lyon Cedex 8, France

Glioblastoma multiforme (GBM), a malignant astrocytic WHO grade IV. The median survival time varies between tumour, represents the most frequent tumour of the human 7 and 11 months (Daumas-Duport et al., 1988; Chinot brain. Nevertheless, its molecular pathology is not well and Martin, 1996). The heterogeneous genetic pattern of understood. We utilized the immune system, which glioma cells entails dysfunction of very di€erent contributes to cancer protection, to help identify new or cellular regulation cycles (Biernat et al., 1997; Kleihues GBM-related . By screening a human GBM cDNA and Burger, 2000). As known for several decades, the library with autologous patient serum (SEREX-approach), immune system is able to recognize tumour cells (Foley, we isolated a termed PHF3 (PHD finger 3). 1953; Prehn and Main, 1957). In recent years, the number The gene product of PHF3 is immunogenic in GBM as of known tumour antigens has increased enormously. A tested in an allogenic patient serum screening demonstrat- variety of methods have been developed for identifying ing antibodies in 24 of 39 (61.53%) sera, whereas none of tumour antigens such as SEREX, cloning of T-cells and the 14 healthy persons had antibodies against PHF3. animal transplantation models (Old, 1981; Sahin et al., While previous SEREX studies revealed allogenic anti- 1995; Boon and van der Bruggen, 1996; Suresh, 1996). body responses up to 40%, our results for PHF3 represent Brain tumours, located in an immunologically privileged the highest reported rate for a speci®c antibody response. site, were believed for a long time not to be involved in We show that GBM patients with an antibody response such immune reactions. Today, astrocytes and microglia against PHF3 show signi®cant better survival than are considered to function as antigen presenting cells patients without PHF3-speci®c antibodies. Because the (Shrikant and Benveniste, 1996). However, with astro- amino acid sequence of PHF3 contains a PHD ®nger (also cytes as the essential part of the blood brain barrier, the termed LAP motif), a TFIIS homology, a proline rich blood brain barrier is most likely a€ected in astrocytic region and nuclear localization signals, it supposedly tumours. Antigens can have contact with immune functions as a . A polyclonal antibody competent cells and induce immune responses (Weller generated against PHF3 shows nuclear expression in most et al., 1992; Zinkernagel et al., 1997). investigated formalin-®xed, paran embedded tissues. In Applying a combined immunological and molecular GBM, PHF3 expression is concentrated in cells surround- approach based on the antibody response against ing necroses. Oncogene (2001) 20, 4107 ± 4114. tumour-expressed antigens, we have isolated PHF3, a transcription factor-like gene in GBM whose expressed Keywords: humoral