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




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Arthritis Research and Therapy

, 13:R59

First Online: 08 April 2011Received: 10 January 2011Revised: 04 March 2011Accepted: 08 April 2011

Abstract

IntroductionFolate 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.

MethodsFunctional 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.

ResultsFab 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.

ConclusionsUnlike 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.

AbbreviationsADCCantibody-dependent cell-mediated cytotoxicity

AMLacute myelogenous leukemia

EC50half maximal effective concentration

ELISAenzyme-linked immunosorbent assay

Fabantigen-binding fragment

FITCfluorescein isothiocyanate

FRfolate receptor

FRβfolate receptor beta

GPIglycosylphosphatidylinositol

hFRβhuman folate receptor beta

KDequilibrium dissociation constant

mAbmonoclonal antibody

NKnatural killer

PBMCperipheral blood mononuclear cell

PBSphosphate-buffered saline

PBST0.05% Tween 20 in phosphate-buffered saline

PCFTproton-coupled folate transporter

RFCreduced folate carrier

rFRβrecombinant folate receptor beta

scFvsingle-chain variable fragment.

Electronic supplementary materialThe online version of this article doi:10.1186-ar3312 contains supplementary material, which is available to authorized users.

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Author: Yang Feng - Jiayin Shen - Emily D Streaker - Michael Lockwood - Zhongyu Zhu - Philip S Low - Dimiter S Dimitrov

Source: https://link.springer.com/







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