Major histocompatibility complex genotyping with massively parallel pyrosequencing.
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.
Wiseman
Roger W
RW
Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Karl
Julie A
JA
Bimber
Benjamin N
BN
O'Leary
Claire E
CE
Lank
Simon M
SM
Tuscher
Jennifer J
JJ
Detmer
Ann M
AM
Bouffard
Pascal
P
Levenkova
Natalya
N
Turcotte
Cynthia L
CL
Szekeres
Edward
E
Wright
Chris
C
Harkins
Timothy
T
O'Connor
David H
DH
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005838:Genotype; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000378:metabolism*; D008252:Macaca fascicularis; D008253:Macaca mulatta; D008254:Macaca nemestrina; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D015395:Histocompatibility Antigens Class I; D004247:DNA