@article{1782adfda128412fa6e85d5009f515d3,
title = "Major histocompatibility complex genotyping with massively parallel pyrosequencing",
abstract = "Major histocompatibility complex (MHC) genetics dictate adaptive cellular immune responses, making robust MHC genotyping methods essential for studies of infectious disease, vaccine development and transplantation. Nonhuman primates provide essential preclinical models for these areas of biomedical research. Unfortunately, given the unparalleled complexity of macaque MHCs, existing methodologies are inadequate for MHC typing of these key model animals. Here we use pyrosequencing of complementary DNA-PCR amplicons as a general approach to determine comprehensive MHC class I genotypes in nonhuman primates. More than 500 unique MHC class I sequences were resolved by sequence-based typing of rhesus, cynomolgus and pig-tailed macaques, nearly half of which have not been reported previously. The remarkable sensitivity of this approach in macaques demonstrates that pyrosequencing is viable for ultra-high-throughput MHC genotyping of primates, including humans.",
author = "Wiseman, {Roger W.} and Karl, {Julie A.} and Bimber, {Benjamin N.} and O'Leary, {Claire E.} and Lank, {Simon M.} and Tuscher, {Jennifer J.} and Detmer, {Ann M.} and Pascal Bouffard and Natalya Levenkova and Turcotte, {Cynthia L.} and Edward Szekeres and Chris Wright and Timothy Harkins and O'Connor, {David H.}",
note = "Funding Information: Fresh blood or frozen peripheral blood mononuclear cell samples from various macaques were graciously provided by L. Picker (Oregon National Primate Research Center), P. Johnson (New England Primate Research Center), D. Read (Battelle Biomedical Research Center), N. Miller (US National Institute of Allergy and Infectious Diseases), J. Hoxie (University of Pennsylvania), J. Mankowski (Johns Hopkins University), T. Andrus (Charles River Biomedical Research Foundation, Inc.) and I. Lussier (Alpha Genesis, Inc.). L. Hetrick, A. Lane, E. Vlach and J. Thimmapuram provided outstanding emulsion PCR, pyrosequencing and informatics support at the University of Illinois at Urbana-Champaign. We thank D. Watkins and R. DeMars for insightful comments on this manuscript. This work was supported by US National Institute of Allergy and Infectious Diseases contract number HHSN266200400088C/N01-AI-40088. Some support was also provided by a subcontract from the Battelle Biomedical Research Center under US National Institute of Allergy and Infectious Diseases contract N01-A1-30061. This publication was made possible in part by grant numbers P51 RR000167 and P40 RR019995 from the US National Center for Research Resources, a component of the US National Institutes of Health to the Wisconsin National Primate Research Center, University of Wisconsin-Madison. This research was conducted in part at a facility constructed with support from Research Facilities Improvement Program grant numbers RR15459-01 and RR020141-01.",
year = "2009",
month = nov,
doi = "10.1038/nm.2038",
language = "English (US)",
volume = "15",
pages = "1322--1326",
journal = "Nature medicine",
issn = "1078-8956",
publisher = "Nature Publishing Group",
number = "11",
}