SSOP TYPING OF THE 10TH INTERNATIONAL HISTOCOMPATIBILITY WORKSHOP REFERENCE CELL-LINES FOR HLA-C ALLELES

HLA-C gene products are the most poorly understood of the HLA class I molecules because they express at low level on the cell surface compared to HLA-A and -B. However, recent evidence shows that HLAC molecules are functionally competent in eliciting T-cell responses and in controlling NK-cell recognition. Approximately 20 to 50% of HLA-C alleles type ''blank'' in most populations. To provide a better definition of the HLA-C alleles, we analyzed 98 extensively characterized B-cell lines from the 10th International Histocompatibility Workshop. Selective HLA-C-specific DNA amplification of exons 2 and 3 from DNA prepared from the cell panel was achieved with the use of two sets of locus-specific primers. We used 64 sequence-specific oligonucleotide probes (SSOPs) complementary to variable sites in exons 2 and 3 to generate hybridization patterns. Twenty-five alleles were found among these patterns, including seven new alleles in the homozygous cell lines and seven potential new alleles in heterozygous cell lines. Differences between the new alleles and known alleles were generally small. Five major groups were identified in the Cw ''blank'' cells by the SSOP patterns. In addition, linkage between HLA-B specificities and HLA-C alleles was similar to previous observations. The present study demonstrated that SSOP typing was effective in identifying new alleles in homozygous typing cells but not in the heterozygous cells. Also, DNA typing can facilitate the identification of all HLA-C alleles, including those that serologically type as blanks. The HLA-C locus may be more polymorphic than was previously recognized.