Nitrocellulose particles adsorbed to immunoglobulins are a new and effective approach to induce cell activation dependent on receptor aggregation.
Nitrocellulose (NC) has proved to be a versatile tool for the isolation and characterization of various biomolecules. In this report we extend its scope by using antibody-coated NC particles to cross-link molecules on the surface of living cells. Ligation of receptors in Jurkat cells with NC-bound specific antibodies induced protein tyrosine phosphorylation patterns of cellular proteins comparable to conventional antibody cross-linking. In addition, the present study shows that application of NC particles coated with human IgA significantly activated monocytic cells via the Fc alpha receptor (Fc alphaR), whereas cross-linking of receptor-ligand complexes with isotype-specific antibody was less efficient. Subsequent immunoprecipitation and immunoblot analysis of aggregated Fc receptors (FcRs) complexed to Ig-adsorbed particles permits fast identification of molecules involved in the transmission of signals. Therefore, ligand-coated NC particles can be used to examine receptor-mediated cell activation events dependent upon extensive receptor aggregation.
Gulle
H
H
Institute of Immunology, University of Vienna, Austria. heinz.gulle@univie.ac.at
Eibl
M M
MM
Wolf
H M
HM
D000327:Adsorption; D003101:Collodion / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D019169:Jurkat Cells / Q000187:drug effects / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects* / Q000502:physiology; D010316:Particle Size; D010766:Phosphorylation / Q000187:drug effects; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011940:Receptor Aggregation / Q000187:drug effects* / Q000502:physiology; D011948:Receptors, Antigen, T-Cell / Q000187:drug effects / Q000502:physiology; D011961:Receptors, Fc / Q000187:drug effects / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D011948:Receptors, Antigen, T-Cell; D011961:Receptors, Fc; D014443:Tyrosine; D003101:Collodion; D011505:Protein-Tyrosine Kinases