RESUMO
A cDNA encoding the human beta Fc gamma receptor II (FcRII) was isolated from a placental cDNA library. Analysis of the predicted amino acid sequence indicates that this receptor is synthesized with a 42-amino acid leader sequence. The mature protein consists of 249 amino acids. The leader sequence and the cytoplasmic domain are strikingly different from the CDw32 antigen but show great homology to the mouse beta 2FcR. RNA blot analysis of human cells using CDw32 and beta FcRII-specific DNA fragments demonstrated one beta FcRII transcript (1.7 kb) in B cells and in HL-60 cells which were induced to differentiate along a monocyte-macrophage pathway by phorbol 12-myristate 13-acetate treatment. Under these conditions the CDw32 transcripts (2.5 and 1.7 kb) are induced to a minor extent in HL-60 cells. In contrast, the 2.5-kb CDw32 transcript is strongly induced in HL-60 cells which have been induced to differentiate into granulocytes by exposure to dimethylsulfoxide. To determine the biological properties of the beta FcRII, we expressed the antigen in FcR- hamster cells. Only immune complexes but not monomeric human IgG were bound significantly. Bound ligand was efficiently internalized within 15 min and it was then found in vesicular structures. Thus the low-affinity beta FcRII is able to internalize ligands without cooperation with any other FcR.
Assuntos
Antígenos de Diferenciação/genética , Receptores Fc/genética , Sequência de Aminoácidos , Animais , Complexo Antígeno-Anticorpo/metabolismo , Antígenos CD/biossíntese , Antígenos CD/genética , Antígenos de Diferenciação/biossíntese , Linfócitos B/metabolismo , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Endocitose/imunologia , Regulação da Expressão Gênica/imunologia , Biblioteca Genômica , Humanos , Dados de Sequência Molecular , Monócitos/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores Fc/biossíntese , Receptores de IgG , Homologia de Sequência do Ácido Nucleico , TransfecçãoRESUMO
Genomic EMBL 3 DNA clones representing part of the human Fc gamma receptor II and the beta Fc gamma receptor II genes were characterized. One of them contains the first five exons including the 5' flanking region of the beta FcRII gene. The signal peptide, the extracellular domains, the putative membrane spanning region, and the first amino acids of the cytoplasmic region encoded by these five exons are spread over approximately 11 kb. Another genomic DNA clone comprises four exons encoding the second extracellular domain and the transmembrane and cytoplasmic regions of the FcRII. Alignment of the genomic DNA clones reveals that these FcR genes are identically organized. Comparison of the corresponding regions of these clones shows that not only the exons are strikingly homologous but also the splice junctions and parts of the intervening sequences are conserved. Furthermore, the genomic organization of FcRII and HLA-class I resemble each other.
Assuntos
Antígenos de Diferenciação/genética , Genes/imunologia , Receptores Fc/genética , Sequência de Bases , Southern Blotting , Genes MHC Classe I/imunologia , Genes MHC da Classe II/imunologia , Humanos , Dados de Sequência Molecular , Receptores de IgG , Mapeamento por RestriçãoRESUMO
Binding of aggregated human immunoglobulin G (IgG) on diploid human fibroblasts leads to a rapid depolarization of the cells within 1-2 min. We resolved this membrane potential change into its plasma membrane and mitochondrial membrane components by measuring the transmembrane distribution of the lipophilic tritium-labelled cation tetraphenylphosphonium, [3H]Ph4P+. The responsibility of the plasma membrane for the membrane potential change, induced by binding of IgGs, is demonstrated. The IgG-induced membrane depolarization leads to the induction of prostaglandin E2 synthesis. Aggregated immunoglobulins (IgG) are specifically bound via the Fc portion because only binding of Fc fragments, in contrast to (Fab')2 fragments, leads to a stimulation of prostaglandin E2 synthesis comparable to that mediated by IgGs. Depolarization of the plasma membrane by short incubation of the fibroblasts in high-K+ buffer (5 min) results in a stimulation of prostaglandin E2 synthesis comparable to that mediated by either aggregated human IgGs or Fc fragments. Our previous results on Fc gamma-receptor-mediated antigen-IgG-antibody complex internalization showed that a maximum uptake of these complexes could be detected 60-90 min after binding. Therefore, we conclude that not internalisation but binding of aggregated IgGs to the Fc gamma receptors on human fibroblasts is the stimulus for plasma membrane depolarization leading to an enhanced prostaglandin E2 release.