Expression of LA45 reactive β2‐microglobulin free HLA Class I α‐chains on activated T‐cells is regulated by internalization, constitutive and protein kinase C inducible release

Abstract

HLA Class I molecules on activated T cells are expressed as mAb W6/32 reactive heterodimers associated with beta 2-microglobulin (beta 2-m) and also as mAb LA45 reactive beta 2-m free HLA Class I alpha-chains. However, the regulation of free alpha-chain expression remained enigmatic. Here we show, that the amount of cell surface expressed free heavy chains is influenced by two distinct mechanisms. Firstly, a proportion of expressed molecules are cleaved and give rise to a soluble pool of HLA Class I molecules. We provide evidence that, besides the previously described constitutive release of free alpha chains, a second phorbol ester inducible release mechanism involving activation of protein kinase C (PKC) does exist. We demonstrate that both the constitutive and the enhanced release of LA45 reactive HLA Class I alpha-chains are the consequence of a cell membrane bound proteolytic activity with the characteristics of a 1, 10 phenanthroline sensitive metalloprotease. Secondly, we report that a distinct fraction of mAb tagged free alpha-chains is internalized via an n-ethylmaleimide sensitive pathway. Together, this data suggests that the expression of free alpha-chains is regulated by pathways governing release and internalization.