Nramp1 transfection transfers Ity/Lsh/Bcg-related pleiotropic effects on macrophage activation: influence on antigen processing and presentation

Abstract

The natural resistance-associated macrophage protein (Nramp1) regulates macrophage activation. One of its pleiotropic effects on macrophage function is to regulate expression of major histocompatibility class II molecules. In this study macrophages stably transfected with the wild-type (infection-resistant) or the natural mutant (infection-susceptible) allele of the Nramp1 gene were used to study class II expression and processing and presentation of recombinant protein antigens to CD4+ T-cell hybridomas. As demonstrated previously for macrophages from Nramp1-resistant and -susceptible congenic mouse strains, transfected macrophage clones carrying the wild-type allele showed enhanced upregulation of class II molecules in response to gamma interferon compared to that shown by macrophage clones carrying an endogenous mutant allele or transfected with the mutant allele expressed under a viral long terminal repeat promoter. The wild-type allele-transfected macrophage clones also demonstrated an enhanced, lipopolysaccharide-dependent ability to process the recombinant leishmanial antigen LACK-delta 1 (the Leishmania homolog of receptors for activated C kinase) for presentation to LACK-specific CD4+ T cells. An influence on antigen processing must therefore be added to the growing list of pleiotropic effects of the Nramp1 gene potentially contributing to its role in infectious and autoimmune disease susceptibility. These results also have important implications for analysis of T-cell responses to vaccination, especially where antigens are presented to the immune system using live Salmonella species or Mycobacterium bovis BCG as a vaccine vehicle.