Evidence implicating I region-restricted antigen presentation in alloantigen and nominal antigen recognition by a dual-reactive helper T lymphocyte clone.

Abstract

A helper T lymphocyte clone, designated A10, was generated from spleen cells of a B10.A mouse and demonstrated reactivity to both the nominal protein antigen hen egg ovalbumin (OVA) presented by I-Ak-bearing antigen-presenting cells (APC) and irradiated I-As-bearing spleen cells in the absence of OVA. A stimulatory signal was delivered to the cloned cells by syngeneic spleen cells that were exposed to OVA and then fixed with paraformaldehyde. However, paraformaldehyde-fixed allogeneic spleen cells that bear the I-As determinant recognized by a monoclonal antibody (mAb) were unable, by themselves, to stimulate the A10 cells. The inability of fixed allogeneic spleen cells to stimulate was rectified by the addition of irradiated I-Ak-positive spleen cells, suggesting that at least in this situation, alloantigen must be presented to the alloreactive A10 cells. Further evidence supporting this proposal for class II-restricted presentation of alloantigen included the observations that 1) irradiated I-Ak-negative spleen cells were unable to present the fixed H-2s spleen cells, 2) the anti-I-As mAb blocked the stimulatory signals delivered by irradiated H-2s spleen cells and by fixed H-2s spleen cells plus irradiated syngeneic spleen cells, 3) some anti-I-Ak mAb preparations were able to inhibit stimulation by OVA and by fixed H-2s spleen cells plus irradiated syngeneic spleen cells, and 4) an anti-L3T4a mAb effectively blocked all three stimulatory signals. These data suggested that alloreactivity can be mediated by an antigen-presentation process similar to that proposed for nominal peptide antigen presentation, and that alloantigen in the form of paraformaldehyde-fixed allogeneic spleen cells is recognized in the context of self-determinants before a stimulatory signal can be delivered.