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Characterization and Expression of CT45 in Hodgkin's Lymphoma

Hans-Jürgen Heidebrecht¹, Alexander Claviez², Marie Luise Kruse³, Marc Pollmann¹, Friedrich Buck⁴, Sönke Harder⁴, Markus Tiemann¹, Wolfgang Dörffel⁵ and Reza Parwaresch^1,

Authors' Affiliations: ¹ Department of Hematopathology and Lymph Node Registry, ² Department of Pediatrics, and ³ First Department of Medicine, University of Kiel, Kiel, Germany; ⁴ Institute for Clinical Chemistry, University of Hamburg, Hamburg, Germany; and ⁵ Second Department of Pediatrics, HELIOS-Klinikum Berlin-Buch, Berlin, Germany

Requests for reprints: Hans-Jürgen Heidebrecht, Department of Hematopathology, University of Kiel, Niemannsweg 11, D-24105 Kiel, Germany. Phone: 49-431-597-3393; Fax: 49-431-597-3428; E-mail: hheidebrecht{at}path.uni-kiel.de.

Purpose: The monoclonal antibody Ki-A10 (IgG1) generated after immunization of mice with Hodgkin's lymphoma cell line L428 detects a nuclear antigen in human tissues with a restricted distribution pattern similar to cancer/testis antigens. The aim of this study was to characterize the antigen and to determine the expression profile in Hodgkin's lymphoma.

Experimental Design: The half-life and phosphorylation of the antigen were determined by radiolabeling. The antigen was characterized by immunopurification and sequencing. Demethylation of genes is used to induce cancer/testis antigens. Ki-A10-negative cells were treated with 5-aza-2'-deoxycytidine. The Ki-A10 expression in paraffin-embedded tumors was determined immunohistochemically.

Results: Immunopurification of the 25/22-kDa antigen and sequencing revealed a peptide of 14 amino acids corresponding to the gene product of the newly described gene family MGC27005, located on chromosome Xq26.3, now termed CT45. CT45 is significantly phosphorylated and down-regulated during mitosis. Demethylation of CT45-negative HeLa cells and stimulated peripheral blood lymphocytes induced CT45 expression. Except testis, immunohistochemical stainings of normal tissues, reactive lymphoid lesions, and most malignant tumors were negative. In comparison, 54 of 99 (55%) samples from pediatric and adolescent Hodgkin's lymphoma patients enrolled in the multicenter trial HD-95 stained Ki-A10 positive. Ki-A10 expression correlated with histologic subtypes (nodular sclerosis Hodgkin's lymphoma 68% versus mixed cellularity Hodgkin's lymphoma 40% versus nodular lymphocyte predominant Hodgkin's lymphoma 9%; P < 0.001).

Conclusions: Ki-A10 is the first monoclonal antibody that detects CT45. As benign lymphoid lesions did not express CT45, the use of Ki-A10 antibody will facilitate the discrimination of Hodgkin's lymphoma from reactive lymphadenopathies.

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