Monoclonal antibodies against the human interleukin-11 receptor alpha-chain (IL-11Ralpha) and their use in studies of human mononuclear cells.
A panel of 14 hybridoma cell lines secreting monoclonal antibodies against the human interleukin-11 receptor alpha chain (hIL-11Ralpha) 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-11Ralpha. Another group of 12 antibodies was generated against a hybrid protein consisting of the extracellular part of the hIL-11Ralpha fused to mature full-length human IL-2. All these antibodies recognized native hIL-11Ralpha and most also recognized the denatured receptor on immunoblots after SDS-PAGE. Four different epitopes were identified on the extracellular part of the hIL-11Ralpha. 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-11Ralpha expressed in gp130/hIL-11Ralpha-co-transfected Ba/F3 cells. The E27 antibody cross-reacted with murine IL-11Ralpha, 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-11Ralpha. This region, which is less conserved between mouse and human, is shown here to be an immunodominant region. Anti-IL-11Ralpha monoclonal antibodies, which have not been described previously enabled us to explore the expression and tissue distribution of IL-11Ralpha 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.
Blanc
C
C
Immunotech: A Beckman-Coulter Company, Marseille, France.
Vusio
P
P
Schleinkofer
K
K
Boisteau
O
O
Pflanz
S
S
Minvielle
S
S
Grötzinger
J
J
Müller-Newen
G
G
Heinrich
P C
PC
Jacques
Y
Y
Montero-Julian
F A
FA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D003429:Cross Reactions; D000939:Epitopes; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D053703:Interleukin-11 Receptor alpha Subunit; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology; D007963:Leukocytes, Mononuclear; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D018123:Receptors, Interleukin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D053702:Receptors, Interleukin-11; D011993:Recombinant Fusion Proteins / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D020349:Surface Plasmon Resonance; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000939:Epitopes; C508428:IL11RA protein, human; C508429:Il11ra1 protein, mouse; C508431:Il11ra2 protein, mouse; D053703:Interleukin-11 Receptor alpha Subunit; D007376:Interleukin-2; D010446:Peptide Fragments; D018123:Receptors, Interleukin; D053702:Receptors, Interleukin-11; D011993:Recombinant Fusion Proteins