The Journal of Immunology, Vol 151, Issue 5 2613-2622, Copyright © 1993 by American Association of Immunologists

ARTICLES

The soluble pool of beta 2-microglobulin free HLA class I alpha-chains. Qualitative and quantitative characterization

WF Pickl, O Majdic, I Fae, R Reuschel, W Holter and W Knapp
Institute for Immunology, University of Vienna, Austria.

Previous studies have demonstrated that HLA class I heterodimers are present in plasma and cell culture supernatants. They can be precipitated by mAb the binding of which is dependent on the proper association of the polymorphic alpha-chain with beta 2-microglobulin (beta 2-m). The molecular mass of the alpha-chain ranges from 45 to 35 kDa with a number of intermediate products. We report on the identification of 35-kDa soluble beta 2-m free HLA class I H chains immunoprecipitated by mAb LA45 from cell culture media of activated B and T cells. Furthermore, a peptide-based competitive immunosorbent assay was established to determine the amounts of soluble HLA class I alpha-chains. By means of this assay, we formally proved the specificity of mAb LA45 for a linear epitope on HLA class I H chains centered on residues arginine-asparagine at positions 62 and 63 of the alpha 1-domain. PHA or rIL-2 were identified as efficient stimuli for PBMC leading to the generation of soluble beta 2-m free HLA class I H chains. Testing of cell lines representing distinct stages of hematopoietic differentiation demonstrated a significant correlation between cell surface expression of beta 2-m free HLA class I H chains and amounts of soluble LA45 reactive molecules. However, three of six human T lymphotropic virus type I transfected cell lines, although expressing beta 2-m free H chains, do not generate soluble molecules. Finally, human sera were found to contain considerable amounts of beta 2-m free HLA class I H chains. The average amount of these molecules in sera of individuals with one positive LA45 allele was determined to be 46.9 +/- 38.6 nM/liter.