High-Throughput Genotyping of Over Two Million Samples: Workflow and Allele Frequencies.

MICA and MICB are ligands of the NKG2D receptor and thereby influence NK and T cell activity. gene polymorphisms, expression levels and the amount of soluble MICA/B in the serum have been linked to autoimmune diseases, infections, and cancer. In hematopoietic stem cell transplantation, matching between donor and patient has been correlated with reduced acute and chronic graft-vs.-host disease and improved survival. Hence, we developed an extremely cost-efficient high-throughput workflow for genotyping for newly registered potential stem cell donors. Since mid-2017, we have genotyped over two million samples using NGS amplicon sequencing for exons 2-5. In donors of German origin, * is the most common allele with a frequency of 42.3%. It is followed by * (11.7%) and * (8.8%). The three most common alleles are * (43.9%), * (21.7%), and * (18.9%). In general, is less diverse than and only 6 alleles, instead of 15, account for a cumulative allele frequency of 99.5%. In 0.5% of the samples we observed at least one allele of or which has so far not been reported to the IPD/IMGT-HLA database. By providing typed voluntary donors, clinicians become empowered to include into their donor selection process to further improve unrelated hematopoietic stem cell transplantation.