Monoclonal antibodies against the human interleukin-11 receptor alpha-chain (IL-11Rα) and their use in studies of human mononuclear cells

• Published in: Journal of immunological methods 2000-07, Vol.241 (1), p.43-59
• Main Authors: Blanc, Chrystel, Vusio, Patricia, Schleinkofer, Karin, Boisteau, Olivier, Pflanz, Stefan, Minvielle, Stéphane, Grötzinger, Joachim, Müller-Newen, Gerhard, Heinrich, Peter C, Jacques, Yannick, Montero-Julian, Félix A
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal; Antibody Affinity; Biological and medical sciences; Cross Reactions; Cytokine receptors; Epitopes; Flow Cytometry - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; interleukin 11 receptors; Interleukin-11 Receptor alpha Subunit; Interleukin-11, Interleukin-11 receptor; Interleukin-2 - genetics; Interleukin-2 - immunology; Leukocytes, Mononuclear; Mice; Molecular immunology; Molecular Sequence Data; Monoclonal antibody; Peptide Fragments - immunology; Receptors, Interleukin - genetics; Receptors, Interleukin - immunology; Receptors, Interleukin - isolation & purification; Receptors, Interleukin-11; Recombinant Fusion Proteins - immunology; Sequence Homology, Amino Acid; Species Specificity; Surface Plasmon Resonance; Techniques; Tissue Distribution
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(00)00194-0

A panel of 14 hybridoma cell lines secreting monoclonal antibodies against the human interleukin-11 receptor alpha chain (hIL-11Rα) was obtained using two different approaches. Two antibodies were raised against peptides of the N- and C-terminal sequences, respectively, of the extracellular part of the hIL-11Rα. Another group of 12 antibodies was generated against a hybrid protein consisting of the extracellular part of the hIL-11Rα fused to mature full-length human IL-2. All these antibodies recognized native hIL-11Rα and most also recognized the denatured receptor on immunoblots after SDS–PAGE. Four different epitopes were identified on the extracellular part of the hIL-11Rα. One epitope, defined by the E27 antibody, is located at the N-terminus and the other three epitopes are clustered in the membrane-proximal, C-terminal region. The antibodies defining epitopes I and II recognized membrane-bound hIL-11Rα expressed in gp130/hIL-11Rα-co-transfected Ba/F3 cells. The E27 antibody cross-reacted with murine IL-11Rα, in agreement with the fact that the N-terminal region is highly conserved between species. The other 13 antibodies all recognized a region between amino acids 319 and 363, which is the membrane-proximal part of the hIL-11Rα. This region, which is less conserved between mouse and human, is shown here to be an immunodominant region. Anti-IL-11Rα monoclonal antibodies, which have not been described previously enabled us to explore the expression and tissue distribution of IL-11Rα on human peripheral blood mononuclear cells and cell lines. The antibodies provide powerful tools for the study of the regulation and function of the receptor.