title: A folate receptor beta-specific human monoclonal antibody recognizes activated macrophage of rheumatoid patients and mediates antibody-dependent cell-mediated cytotoxicity.
abstract: BACKGROUND
  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.
authors.affiliation: Protein Interactions Group, CCRNP, NCI-Frederick, NIH, 1050 Boyle Street, Frederick, MD 21702, USA. fengya@mail.nih.gov
authors.forename: Yang
authors.initials: Y
authors.lastname: Feng
authors.forename: Jiayin
authors.initials: J
authors.lastname: Shen
authors.forename: Emily D
authors.initials: ED
authors.lastname: Streaker
authors.forename: Michael
authors.initials: M
authors.lastname: Lockwood
authors.forename: Zhongyu
authors.initials: Z
authors.lastname: Zhu
authors.forename: Philip S
authors.initials: PS
authors.lastname: Low
authors.forename: Dimiter S
authors.initials: DS
authors.lastname: Dimitrov
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D002469:Cell Separation; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D058976:Folate Receptor 2 / Q000276:immunology*; D006801:Humans; D008262:Macrophage Activation / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D018613:Microscopy, Confocal; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D058976:Folate Receptor 2; D019151:Peptide Library; D011994:Recombinant Proteins