title: Sequence capture and next generation resequencing of the MHC region highlights potential transplantation determinants in HLA identical haematopoietic stem cell transplantation.
abstract: How cells coordinate the immune system activities is important for potentially life-saving organ or stem cell transplantations. Polymorphic immunoregulatory genes, many of them located in the human major histocompatibility complex, impact the process and assure the proper execution of tolerance-versus-activity mechanisms. In haematopoietic stem cell transplantation, on the basis of fully human leukocyte antigen (HLA)-matched donor-recipient pairs, adverse effects like graft versus leukaemia and graft versus host are observed and difficult to handle. So far, high-resolution HLA typing was performed with Sanger sequencing, but for methodological reasons information on additional immunocompetent major histocompatibility complex loci has not been revealed. Now, we have used microarray sequence capture and targeted enrichment combined with next generation pyrosequencing for 3.5 million base pair human major histocompatibility complex resequencing in a clinical transplant setting and describe 3025 variant single nucleotide polymorphisms, insertions and deletions among recipient and donor in a single sequencing experiment. Taken together, the presented data show that sequence capture and massively parallel pyrosequencing can be used as a new tool for risk assessment in the setting of allogeneic stem cell transplantation.
authors.affiliation: Red Cross Transfusion Service for Upper Austria, Linz. johannes.proell@o.roteskreuz.at
authors.forename: Johannes
authors.initials: J
authors.lastname: Pröll
authors.forename: Martin
authors.initials: M
authors.lastname: Danzer
authors.forename: Stephanie
authors.initials: S
authors.lastname: Stabentheiner
authors.forename: Norbert
authors.initials: N
authors.lastname: Niklas
authors.forename: Christa
authors.initials: C
authors.lastname: Hackl
authors.forename: Katja
authors.initials: K
authors.lastname: Hofer
authors.forename: Sabine
authors.initials: S
authors.lastname: Atzmüller
authors.forename: Peter
authors.initials: P
authors.lastname: Hufnagl
authors.forename: Christian
authors.initials: C
authors.lastname: Gülly
authors.forename: Hanns
authors.initials: H
authors.lastname: Hauser
authors.forename: Otto
authors.initials: O
authors.lastname: Krieger
authors.forename: Christian
authors.initials: C
authors.lastname: Gabriel
mesh_terms: D000939:Epitopes / Q000235:genetics*; D006086:Graft vs Host Disease / Q000235:genetics; D006680:HLA Antigens / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D059014:High-Throughput Nucleotide Sequencing; D006650:Histocompatibility Testing; D006801:Humans; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D000939:Epitopes; D006680:HLA Antigens