Immunohistochemical characterization of intracellular J-chain and binding site for secretory component (SC) in human immunoglobulin (Ig)-producing cells

Abstract

J-chain staining of IgA- and IgM-producing immunocytes was significantly enchanced when tissue sections were pretreated with acid urea, apparently because molecular unfolding exposed concealed J-chains. This indicated substantial completion of the Ig polymers at the cytoplasmic level, which was verified by diffuse binding of SC in vitro to the cytoplasm of most J-chain-positive IgA and IgM cells. This process involved specific non-covalent forces which showed the same interrelation as that noted for isolated dimeric IgA and 19S IgM-the latter as well as IgM cells exhibiting stronger binding of SC than the IgA counterparts. Conversely, J-chain staining of IgD and IgG immunocytes was not enhanced by acid urea and these cells did not generally express affinity for SC; rare exceptions could apparently be ascribed to artifacts or dual isotype production including IgA or IgM polymers. Parallel demonstration of J-chain and SC binding seems to be the best available method for studies of polymer-producing immunocyte populations and offers the advantage of in situ evaluation of cell distribution in relation to morphology. The reliability of this approach was attested to by the fact that IgA immunocytes in all secretory tissues investigated (salivary, mammary and lacrimal glands; nasal and intestinal mucosae) expressed J-chain (87–97%) and SC affinity (84–87%) in comparable proportions, indicating that almost 90% of the cells were engaged mainly in dimer production. The observation that most IgD and 50–70% of the IgG immunocytes in secretory tissues expressed J-chain, has implications for the differentiation of B-cell clones homing to such sites. Conversely, IgG cells in extra-glandular tissues showed strikingly reduced J-chain production and such sites contained IgA immunocytes with heterogeneous expression of J-chain and SC affinity. Thus, in the extra-follicular area of palatine tonsils 70–80% of the IgA cells seemed to be pure monomer producers and the remainders apparently generated a mixed product. Most immunocytes in extra-glandular tissues may therefore belong to mature clones with completely or partially repressed J-chain synthesis.