Recombinant human single chain Fv antibodies recognizing human interleukin-6. Specific targeting of cytokine-secreting cells.
A human antibody library was displayed on the surface of filamentous bacteriophage and screened for binding to human interleukin-6 (IL-6). Two antibody-bearing phages were selected that bound IL-6. The complementary-determining region 3 loops of the variable heavy chains of these two antibodies differed in length and sequence and recognized two distinct epitopes. One of the single chain Fv fragments isolated (H1) was found to bind human (but not murine) IL-6 with an affinity comparable to that of the human IL-6 receptor. H1 also recognized newly synthesized human IL-6 intracellularly, as shown by indirect immunofluorescence. H1 did not neutralize human IL-6, and the H1 epitope was mapped to a region of IL-6 not involved in interactions with IL-6, IL-6 receptor, or the signal-transducing protein gp130. To target IL-6-secreting cells, we then constructed a bispecific antibody fragment (a diabody) comprising H1 and the antigen binding site of the T-cell activating monoclonal antibody OKT3. The diabody led to T-cell-mediated killing of cells secreting IL-6.
Krebs
B
B
Department of Medicine, Section of Pathophysiology, Johannes Gutenberg-University of Mainz, Obere Zahlbacher Strasse 63, D-55101 Mainz, Germany.
Griffin
H
H
Winter
G
G
Rose-John
S
S
D018033:Antibodies, Bispecific / Q000494:pharmacology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D017252:CD3 Complex / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D018604:Epitope Mapping; D000939:Epitopes; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D000922:Immunotoxins / Q000494:pharmacology*; D015850:Interleukin-6 / Q000276:immunology*; D009500:Neutralization Tests; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology
D018033:Antibodies, Bispecific; D017252:CD3 Complex; D000939:Epitopes; D007128:Immunoglobulin Fragments; D000922:Immunotoxins; D015850:Interleukin-6; D019151:Peptide Library; D011994:Recombinant Proteins; C055966:immunoglobulin Fv