title: RA8, a human anti-CD25 antibody against human Treg cells.
abstract: Although anti-CD25 antibodies exist for clinical use in patients, there is a need for the development of a human Treg antibody that will abrogate the immunosuppressive function of this small but critical T cell subtype. Based upon mounting evidence that the level of Treg cells in the tumor microenvironment correlates with clinical prognosis and stage in man, it appears that Treg cells play an important role in the tumor's ability to overcome host immune responses. In mice, the rat anti-mouse CD25 antibody PC61 causes depletion of CD25-bearing Treg cells both peripherally in lymphatic tissues and in the tumor microenvironment, without inducing symptoms of autoimmunity. A similar antibody, though with the ability to delete Treg cells specifically, would be an important new tool for reversing tumor escape associated with Treg immunosuppression in man. To begin to generate such a reagent, we now describe the development of a human anti-CD25 antibody using a novel yeast display library. The target antigen CD25-Fc was constructed and used for five rounds of selection using a non-immune yeast display library that contained as many as 10(9) single chain variable fragments (scFv). Two unique clones with low K(D) values (RA4 and RA8) were then selected to construct fully human anti-CD25 antibodies (IgG1/kappa) for stable expression. One antibody, RA8, showed excellent binding to human CD25(+) cell lines and to human Treg cells and appears to be an excellent candidate for the generation of a human reagent that may be used in man for the immunotherapy of cancer.
authors.affiliation: Department of Pathology, University of Southern California, Keck School of Medicine, Los Angeles, California.
authors.forename: Robyn S
authors.initials: RS
authors.lastname: Arias
authors.forename: Meg L
authors.initials: ML
authors.lastname: Flanagan
authors.forename: Keith D
authors.initials: KD
authors.lastname: Miller
authors.forename: Yu-Chih
authors.initials: YC
authors.lastname: Nien
authors.forename: Peisheng
authors.initials: P
authors.lastname: Hu
authors.forename: Dixon
authors.initials: D
authors.lastname: Gray
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Khawli
authors.forename: Alan L
authors.initials: AL
authors.lastname: Epstein
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D066298:In Vitro Techniques; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology*; D051379:Mice; D019151:Peptide Library; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007141:Immunoglobulin Fc Fragments; D007135:Immunoglobulin Variable Region; D053645:Interleukin-2 Receptor alpha Subunit; D019151:Peptide Library; D011993:Recombinant Fusion Proteins