Rituximab Induces Different but Overlapping Sets of Genes in Human B-lymphoma Cell Lines

Overview

Journal Cancer Immunol Immunother

Specialties Allergy & Immunology
Oncology
Pharmacology

Date 2004 Sep 28

PMID 15449038

Citations 7

Authors

Elena Cittera

Chiara Onofri

Maria dApolito

Guillaume Cartron

Giovanni Cazzaniga

Leopoldo Zelante

Paolo Paolucci

Andrea Biondi

Martino Introna

Josee Golay

Affiliations

Soon will be listed here.

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 demonstrate 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.

Citing Articles

Protective effects of rituximab on puromycin-induced apoptosis, loss of adhesion and cytoskeletal alterations in human podocytes.

Jeruschke S, Alex D, Hoyer P, Weber S Sci Rep. 2022; 12(1):12297.

PMID: 35853959 PMC: 9296604. DOI: 10.1038/s41598-022-16333-w.

Cytoplasmic location of NR4A1 in aggressive lymphomas is associated with a favourable cancer specific survival.

Fechter K, Feichtinger J, Prochazka K, Unterluggauer J, Pansy K, Steinbauer E Sci Rep. 2018; 8(1):14528.

PMID: 30266952 PMC: 6162226. DOI: 10.1038/s41598-018-32972-4.

Rituximab targets podocytes in recurrent focal segmental glomerulosclerosis.

Fornoni A, Sageshima J, Wei C, Merscher-Gomez S, Aguillon-Prada R, Jauregui A Sci Transl Med. 2011; 3(85):85ra46.

PMID: 21632984 PMC: 3719858. DOI: 10.1126/scitranslmed.3002231.

Antibodies against CD20 or B-cell receptor induce similar transcription patterns in human lymphoma cell lines.

Franke A, Niederfellner G, Klein C, Burtscher H PLoS One. 2011; 6(2):e16596.

PMID: 21364752 PMC: 3041769. DOI: 10.1371/journal.pone.0016596.

γδ T-cell killing of primary follicular lymphoma cells is dramatically potentiated by GA101, a type II glycoengineered anti-CD20 monoclonal antibody.

Braza M, Klein B, Fiol G, Rossi J Haematologica. 2010; 96(3):400-7.

PMID: 21109686 PMC: 3046271. DOI: 10.3324/haematol.2010.029520.

References

Semac I, Palomba C, Kulangara K, Klages N, van Echten-Deckert G, Borisch B . Anti-CD20 therapeutic antibody rituximab modifies the functional organization of rafts/microdomains of B lymphoma cells. Cancer Res. 2003; 63(2):534-40. View

Kawahara K, Oyadomari S, Gotoh T, Kohsaka S, Nakayama H, Mori M . Induction of CHOP and apoptosis by nitric oxide in p53-deficient microglial cells. FEBS Lett. 2001; 506(2):135-9. DOI: 10.1016/s0014-5793(01)02898-8. View

Liu G, Loraine A, Shigeta R, Cline M, Cheng J, Valmeekam V . NetAffx: Affymetrix probesets and annotations. Nucleic Acids Res. 2003; 31(1):82-6. PMC: 165568. DOI: 10.1093/nar/gkg121. View

Shan D, Ledbetter J, Press O . Apoptosis of malignant human B cells by ligation of CD20 with monoclonal antibodies. Blood. 1998; 91(5):1644-52. View

Cartron G, Dacheux L, Salles G, Solal-Celigny P, Bardos P, Colombat P . Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcgammaRIIIa gene. Blood. 2002; 99(3):754-8. DOI: 10.1182/blood.v99.3.754. View

Fambrough D, McClure K, Kazlauskas A, Lander E . Diverse signaling pathways activated by growth factor receptors induce broadly overlapping, rather than independent, sets of genes. Cell. 1999; 97(6):727-41. DOI: 10.1016/s0092-8674(00)80785-0. View

Crittenden M, Gough M, Harrington K, Olivier K, Thompson J, Vile R . Expression of inflammatory chemokines combined with local tumor destruction enhances tumor regression and long-term immunity. Cancer Res. 2003; 63(17):5505-12. View

Coiffier B, Lepage E, Briere J, Herbrecht R, Tilly H, Bouabdallah R . CHOP chemotherapy plus rituximab compared with CHOP alone in elderly patients with diffuse large-B-cell lymphoma. N Engl J Med. 2002; 346(4):235-42. DOI: 10.1056/NEJMoa011795. View

Glynne R, Akkaraju S, Healy J, Rayner J, Goodnow C, Mack D . How self-tolerance and the immunosuppressive drug FK506 prevent B-cell mitogenesis. Nature. 2000; 403(6770):672-6. DOI: 10.1038/35001102. View

10.

Deans J, Li H, Polyak M . CD20-mediated apoptosis: signalling through lipid rafts. Immunology. 2002; 107(2):176-82. PMC: 1782791. DOI: 10.1046/j.1365-2567.2002.01495.x. View

11.

Tedder T, Engel P . CD20: a regulator of cell-cycle progression of B lymphocytes. Immunol Today. 1994; 15(9):450-4. DOI: 10.1016/0167-5699(94)90276-3. View

12.

Edwards J, Leandro M, Cambridge G . B-lymphocyte depletion therapy in rheumatoid arthritis and other autoimmune disorders. Biochem Soc Trans. 2002; 30(4):824-8. DOI: 10.1042/bst0300824. View

13.

Krzysiek R, Lefevre E, Zou W, Foussat A, Bernard J, Portier A . Antigen receptor engagement selectively induces macrophage inflammatory protein-1 alpha (MIP-1 alpha) and MIP-1 beta chemokine production in human B cells. J Immunol. 1999; 162(8):4455-63. View

14.

Nomiyama H, Hieshima K, Hirokawa K, Hattori T, Takatsuki K, Miura R . Characterization of cytokine LD78 gene promoters: positive and negative transcriptional factors bind to a negative regulatory element common to LD78, interleukin-3, and granulocyte-macrophage colony-stimulating factor gene promoters. Mol Cell Biol. 1993; 13(5):2787-801. PMC: 359660. DOI: 10.1128/mcb.13.5.2787-2801.1993. View

15.

Chan H, Hughes D, French R, Tutt A, Walshe C, Teeling J . CD20-induced lymphoma cell death is independent of both caspases and its redistribution into triton X-100 insoluble membrane rafts. Cancer Res. 2003; 63(17):5480-9. View

16.

Clynes R, Towers T, Presta L, Ravetch J . Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor targets. Nat Med. 2000; 6(4):443-6. DOI: 10.1038/74704. View

17.

Golay J, Zaffaroni L, Vaccari T, Lazzari M, Borleri G, Bernasconi S . Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis. Blood. 2000; 95(12):3900-8. View

18.

Deans J, Schieven G, Shu G, VALENTINE M, Gilliland L, Aruffo A . Association of tyrosine and serine kinases with the B cell surface antigen CD20. Induction via CD20 of tyrosine phosphorylation and activation of phospholipase C-gamma 1 and PLC phospholipase C-gamma 2. J Immunol. 1993; 151(9):4494-504. View

19.

Polyak M, Deans J . Alanine-170 and proline-172 are critical determinants for extracellular CD20 epitopes; heterogeneity in the fine specificity of CD20 monoclonal antibodies is defined by additional requirements imposed by both amino acid sequence and quaternary.... Blood. 2002; 99(9):3256-62. DOI: 10.1182/blood.v99.9.3256. View

20.

Petrie R, Deans J . Colocalization of the B cell receptor and CD20 followed by activation-dependent dissociation in distinct lipid rafts. J Immunol. 2002; 169(6):2886-91. DOI: 10.4049/jimmunol.169.6.2886. View