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Rituximab induces different but overlapping sets of genes in human B-lymphoma cell lines

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Abstract

The therapeutic unconjugated anti-CD20 Mab rituximab is used for the treatment of B-non-Hodgkin’s lymphomas. We have studied the direct biological effects, signalling and gene expression profiles induced by rituximab in two human B-lymphoma cell lines, DHL4 and BJAB, using microarray, quantitative PCR and gel shift analysis. Rituximab alone inhibited thymidine uptake and induced homotypic adhesion in DHL4 only, but not BJAB. Analysis of Affymetrix microchips carrying probes for about 10,000 human cDNAs, allowed us to identify 16 genes in DHL4 and 12 in BJAB induced by rituximab at 4 h. Eleven and seven of these genes were specific for DHL4 and BJAB, respectively; whereas the remaining five were up-regulated in both cell lines. Mean induction ranged from 2- to 16-fold. Real time PCR analysis allowed us to confirm up-regulation of all genes identified, except one in BJAB. Time course of induction of eight genes was studied, showing peak induction in most cases at 4 h. The up-regulation of 5/5 genes was also observed with the F(ab′)2 fragment of rituximab. Analysis of three further B-cell lymphoma lines showed that gene induction is not restricted to BJAB and DHL4. Finally, we show that rituximab alone can induce AP1 activation in both cell lines and provide evidence that the ERK1/2 pathway is involved in the rituximab-mediated up-regulation of gene expression. These data demontrate that rituximab alone has direct signalling capacity in different B-lymphoma lines, inducing distinct but overlapping sets of genes which may play a role in the biological and/or therapeutic effect of the antibody.

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Acknowledgements

We thank Prof. E.S. Vitetta (Cancer Immunobiology Cancer, University of Texas Southwestern Medical Center, Dallas, TX, USA) for her generous gift of rituximab F(ab′)2 fragment.This work was supported by the Italian Ministry for University and Research (projects FIRB no. RBAU01 J2ER and no. RBAU01H8SX), the Italian Ministry of Health (ICS PERF00001), the Associazione Italiana Ricerca sul Cancro (AIRC to J.G. and M.I.), Roche Italia (Monza, Italy) and the Istituto Superiore di Sanità (ISS Rome, project 30D.41) and the Associazione Italiana contro le Leucemie (AIL)-sezione Paolo Belli Bergamo.

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Author notes

  1. Martino Introna & Josée Golay

    Present address: Laboratory of Cellular and Gene Therapy “G. Lanzani”, Presidio Matteo Rota, Ospedali Riuniti Bergamo, via Garibaldi 11-13, 24128, Bergamo, Italy

  2. Guillaume Cartron

    Present address: Oncologie Medicale et Maladies du Sang, CHRU Bretonneau, Tours, France

  3. Paolo Paolucci

    Present address: Department of Pediatrics, University of Modena and Reggio Emilia, Modena, Italy

Authors and Affiliations

  1. Laboratory of Molecular Immunohaematology, Department of Immunology, Istituto Ricerche Farmacologiche “Mario Negri”, Milan, Italy

    Elena Cittera, Chiara Onofri, Guillaume Cartron, Martino Introna & Josée Golay

  2. Medical Genetics Service, IRCCS Hospital CSS, Foggia, Italy

    Maria D’Apolito & Leopoldo Zelante

  3. Centro Ricerca M. Tettamanti, Clinica Pediatrica Università di Milano-Bicocca, Monza, Italy

    Giovanni Cazzaniga & Andrea Biondi

  4. Department of Pediatrics, IRCCS Hospital CSS, Foggia, Italy

    Paolo Paolucci

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  1. Elena Cittera
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Correspondence to Josée Golay.

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Cittera, E., Onofri, C., D’Apolito, M. et al. Rituximab induces different but overlapping sets of genes in human B-lymphoma cell lines. Cancer Immunol Immunother 54, 273–286 (2005). https://doi.org/10.1007/s00262-004-0599-4

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