Development of Rituximab-Resistant B-NHL Clones: An In Vitro Model for Studying Tumor Resistance to Monoclonal Antibody-Mediated Immunotherapy

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 731)

Abstract

Therapeutic strategies for cancer include chemotherapy, immunotherapy, and radiation. Such therapies result in significant short-term clinical responses; however, relapses and recurrences occur with no treatments. Targeted therapies using monoclonal antibodies have improved responses with minimal toxicities. For instance, Rituximab (chimeric anti-CD20 monoclonal antibody) was the first FDA-approved monoclonal antibody for the treatment of patients with non-Hodgkin’s lymphoma (NHL). The clinical response was significantly improved when used in combination with chemotherapy. However, a subset of patients does not respond or becomes resistant to further treatment. Rituximab-resistant (RR) clones were used as a model to address the potential mechanisms of resistance. In this chapter, we discuss the underlying molecular mechanisms by which rituximab signals the cells and modifies several intracellular survival/antiapoptotic pathways, leading to its chemo/immunosensitizing activities. RR clones were developed to mimic in vivo resistance observed in patients. In comparison with the sensitive parental cells, the RR clones are refractory to rituximab-mediated cell signaling and chemosensitization. Noteworthy, interference with the hyperactivated survival/antiapoptotic pathways in the RR clones with various pharmacological inhibitors mimicked rituximab effects in the parental cells. The development of RR clones provides a paradigm for studying resistance by other anticancer monoclonal antibodies in various tumor models.

Key words

Rituximab-resistant clones Bcl-2 family Methods of resistance 

Abbreviations

2MAM-A3

2-Methoxyantimycin-A3

ADCC

Antibody-dependent cell-mediated cytotoxicity

AP-1

Activator protein-1

ARL

Acquired immunodeficiency syndrome (AIDS)-related lymphoma

Bcl-2

B-cell lymphoma protein 2

Bcl-xL

Bcl-2 related gene (long alternatively spliced variant of Bcl-x gene)

CDC

Complement-dependent cytotoxicity

DHMEQ

Dehydroxymethylepoxyquinomicin

DLBCL

Diffuse large B-cell lymphoma

ERK1/2 MAPK

Extracellular signal-regulated kinase1/2 mitogen activated protein kinase

FACS

Fluorescence-activated cell sorter

IKK

Inhibitor of kappa B (IκB) kinase complex

Mcl-1

Myeloid cell differentiation 1

RKIP

Raf-1 kinase inhibitor protein

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Surgery, Jonsson Comprehensive Cancer Center, David Geffen School of MedicineUniversity of CaliforniaLos AngelesUSA

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