Antigen processing and presentation by a mouse macrophage-like cell line expressing human HLA class II molecules

Abstract

Macrophages differ from other antigen-presenting cell types in their potential to take up, process and present particulate antigens as well as proteins and peptides. To study their influence on T cell activation we transfected the mouse macrophage-like cell line P388D1 with human genes coding for the α and β chains of class II molecules (DRA(*)0101 and DRB1(*)0401 or DRB1(*)0404) or with a series of localized DR β mutants. The transfected cells (TP cells) all expressed DR4 molecules on their surface, Since they stimulated some, but not all, human DR4-reactive T cell clones, some of the resident peptides are evidently similar in mouse and man. Proteins and polypeptides such as pepsin or the human acetylcholine receptor (AChR) α37-181 were also processed correctly by these transfectants and then presented efficiently to other T cell clones. Nearly all the mutants tested could present to the pepsin-specific clone, establishing functional expression of the transfected DR molecules. For the more exacting AChR α144-156-specific T cell clone PM-A1, we confirmed that the single Gly86 → Val mutation in the DR β chain abolished presentation by two DR4 subtypes. If, however, this same replacement was made in a third variant that has Lys71 (rather than Arg71), the effects were less drastic. This approach could also be used to analyse the contributions of individual substitutions that confer susceptibility to rheumatoid arthritis.