A highly divergent microsatellite facilitating fast and accurate DRB haplotyping in humans and rhesus macaques

^†Department of Comparative Genetics and Refinement, Biomedical Primate Research Centre, P.O. Box 3306, 2280 GH, Rijswijk, The Netherlands; and
^§Department of Immunohematology and Blood Transfusion, Leiden University Medical Centre, E3-Q, P.O. Box 9600, 2300 RC, Leiden, The Netherlands

Contributed by Jon J. van Rood, March 30, 2007 (received for review October 27, 2006)

Abstract

The DRB region of the MHC in primate species is known to display abundant region configuration polymorphism with regard to the number and content of genes present per haplotype. Furthermore, depending on the species studied, the different DRB genes themselves may display varying degrees of allelic polymorphism. Because of this combination of diversity (differential gene number) and polymorphism (allelic variation), molecular typing methods for the primate DRB region are cumbersome. All intact DRB genes present in humans and rhesus macaques appear to possess, however, a complex and highly divergent microsatellite. Microsatellite analysis of a sizeable panel of outbred rhesus macaques, covering most of the known Mamu-DRB haplotypes, resulted in the definition of unique genotyping patterns that appear to be specific for a given haplotype. Subsequent examination of a representative panel of human cells illustrated that this approach also facilitates high-resolution HLA-DRB typing in an easy, quick, and reproducible fashion. The genetic composition of this complex microsatellite is shown to be in concordance with the phylogenetic relationships of various HLA-DRB and Mamu-DRB exon 2 gene/lineage sequences. Moreover, its length variability segregates with allelic variation of the respective gene. This simple protocol may find application in a variety of research avenues such as transplantation biology, disease association studies, molecular ecology, paternity testing, and forensic medicine.

Footnotes

^‡To whom correspondence may be addressed. E-mail: doxiadis{at}bprc.nl or vanrood{at}europdonor.nl
Author contributions: J.J.v.R. and R.E.B. designed research; N.d.G. performed research; F.H.J.C. and I.I.N.D. contributed new reagents/analytic tools; G.G.M.D. analyzed data; and G.G.M.D. and R.E.B. wrote the paper.
The authors declare no conflict of interest.
Data deposition: The sequences reported in this paper have been deposited in the EMBL Nucleotide Sequence Database (accession nos. AM490200–AM490205).
This article contains supporting information online at www.pnas.org/cgi/content/full/0702964104/DC1.
Abbreviation:

STR,

short tandem repeat.
Freely available online through the PNAS open access option.