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Molecular cloning of a macrophage-derived, interferon-inducible secreted immunoglobulin-binding protein
P Nickolaus1, H G Rammensee, R Zawatzky
1Institute for Cell Biology, Department of Immunology, University of Tübingen, Germany. peter.nickolaus@uni-tuebingen.de
We describe the molecular cloning of a 1803-bp cDNA coding for a product termed interferon-induced immunoglobulin-binding protein (IIBP) from a library of IFN-alpha-induced primary bone marrow macrophages. The open reading frame encodes a protein of 26-kDa containing two immunoglobulin-like and one Fc receptor-like domain. Due to the constitutive release of low amounts of IFN-beta, the IIBP mRNA is already present in macrophage-colony-stimulating factor-cultured macrophages. Its expression could be blocked in the presence of either anti-IFN-beta or interleukin-4, which down-regulates the endogenous IFN-beta production. Upon addition of rIFN-alpha4 a 3-5-fold superinduction of IIBP mRNA was observed. Rat monoclonal antibodies detected a protein of the predicted size exclusively in cell culture supernatants of primary bone marrow macrophages and a B-cell line. In immunoprecipitation experiments an unknown 30-kDa protein co-precipitated. The secreted IIBP showed considerable binding to nonspecific rat IgG2a and could be precipitated using mouse IgG2a, IgG2b and IgG3 antibodies of irrelevant specificities, indicating that this gene product is a novel secreted immunoglobulin-binding protein with a new IgG isotype binding pattern that differs to that of known Fc receptors.
We describe the molecular cloning of a 1803-bp cDNA coding for a product termed interferon-induced immunoglobulin-binding protein (IIBP) from a library of IFN-alpha-induced primary bone marrow macrophages. The open reading frame encodes a protein of 26-kDa containing two immunoglobulin-like and one Fc receptor-like domain. Due to the constitutive release of low amounts of IFN-beta, the IIBP mRNA is already present in macrophage-colony-stimulating factor-cultured macrophages. Its expression could be blocked in the presence of either anti-IFN-beta or interleukin-4, which down-regulates the endogenous IFN-beta production. Upon addition of rIFN-alpha4 a 3-5-fold superinduction of IIBP mRNA was observed. Rat monoclonal antibodies detected a protein of the predicted size exclusively in cell culture supernatants of primary bone marrow macrophages and a B-cell line. In immunoprecipitation experiments an unknown 30-kDa protein co-precipitated. The secreted IIBP showed considerable binding to nonspecific rat IgG2a and could be precipitated using mouse IgG2a, IgG2b and IgG3 antibodies of irrelevant specificities, indicating that this gene product is a novel secreted immunoglobulin-binding protein with a new IgG isotype binding pattern that differs to that of known Fc receptors.