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Open Access Research article

A folate receptor beta-specific human monoclonal antibody recognizes activated macrophage of rheumatoid patients and mediates antibody-dependent cell-mediated cytotoxicity

Yang Feng1*, Jiayin Shen2, Emily D Streaker3, Michael Lockwood4, Zhongyu Zhu1, Philip S Low2 and Dimiter S Dimitrov1

Author Affiliations

1 Protein Interactions Group, CCRNP, NCI-Frederick, NIH, 1050 Boyle Street, Frederick, MD 21702, USA

2 Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, IN 47906, USA

3 BRP, SAIC-Frederick, Inc., NCI-Frederick, 1050 Boyle Street, Frederick, MD 21702, USA

4 Indiana University Health Arnett Physicians, 2600 Greenbush Street, Lafayette, IN 47904, USA

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Arthritis Research & Therapy 2011, 13:R59  doi:10.1186/ar3312

Published: 8 April 2011

Abstract

Introduction

Folate receptor beta (FRβ) is only detectable in placenta and limited to some hematopoietic cells of myeloid lineage in healthy people. Studies have indicated that FRβ is over-expressed in activated macrophages in autoimmune diseases and some cancer cells. In this study we aimed to develop an FRβ-specific human monoclonal antibody (mAb) that could be used as a therapeutic agent to treat rheumatoid arthritis and other autoimmune diseases, as well as FRβ positive cancers.

Methods

Functional recombinant FRβ protein was produced in insect cells and used as antigen to isolate a mAb, m909, from a human naïve Fab phage display library. Binding of Fab and IgG1 m909 to FRβ was measured by ELISA, surface plasmon resonance, immune fluorescence staining, and flow cytometry. Antibody-dependent cell-mediated cytotoxicity (ADCC) was evaluated with FRβ positive CHO cells as target cells and isolated peripheral blood monocytes as effector cells in an in vitro assay.

Results

Fab m909 bound with relatively high affinity (equilibrium dissociation constant 57 nM) to FRβ. The IgG1 m909 showed much higher (femtomolar) avidity as measured by ELISA, and it bound to FRβ positive cells in a dose-dependent manner, but not to parental FRβ negative cells. m909 did not compete with folate for the binding to FRβ on cells. m909 was not only able to select FRβ positive, activated macrophages from synovial fluid cells of arthritis patients as efficiently as folate, but also able to mediate ADCC in FRβ positive cells.

Conclusions

Unlike folate-drug conjugates, m909 selectively binds to FRβ, does not recognize FRα, and has at least one effector function. m909 alone has potential to eliminate FRβ positive cells. Because m909 does not compete with folate for receptor binding, it can be used with folate-drug conjugates in a combination therapy. m909 can also be a valuable research reagent.