PCR‐RFLP‐based Mamu‐DQB1 typing of rhesus monkeys: characterization of two novel alleles

Abstract

Up to now 19 allelic sequences of the rhesus monkey DQB1 locus have been published. Referring to these sequences, we have developed a typing protocol for Mamu-DQB1 alleles which was verified by additional cloning, sequence analysis and segregation studies. The protocol is based on the amplification of the second exon with only one specific primer pair followed by the digestion of the PCR products with up to 10 different restriction endonucleases. The alleles can be identified in homozygous and heterozygous combinations since most amplified second exon sequences give unique hand patterns after digestion with at least one of the selected restriction endonucleases. By the use of this protocol we analyzed DNA-samples from 182 rhesus monkeys. Among these samples two novel Mamu-DQB1 alleles were detected, subsequently cloned and their nucleic sequence determined. Since we typed four complete breeding groups consisting of two generations we were able to identify several DQ haplotypes by segregation analysis using the previously developed typing protocol for DQA1.

Characterization of the major histocompatibility complex class II DQB (MhcMamu-DQB1) alleles in a cohort of Chinese rhesus macaques (Macaca mulatta)

“…After years of extensive studies using methods including serologic tests [47], two-dimensional isoelectric focusing [48], polymerase chain reaction-single-strand polymorphism (PCR-SSP) [30], reference strand-mediated conformation analysis [49], PCR-restriction fragment length polymorphism [20], PCR-denaturing gradient gel eletrophoresis [50], and tworounds PCR and sequencing [22], substantial progress has been made in characterizing the MHC in rhesus macaques. Just as in human beings, rhesus macaque MHC class II consists of -DP, -DQ, and -DR regions; however, unlike that in human beings, the MHC in rhesus macaques is much more complex.…”

“…Until now, most reports on rhesus macaque MHC genotypes are based on Indian rhesus macaques, and only about 40 MhcMamu-DQB1 alleles with unique exon 2 sequences have been reported [17][18][19][20][21][22][23][24][25]; limited information is available on the MHC genotypes of Chinese rhesus macaques. Because of the existence of marked differences between the two geographic populations in regard to a wide variety of aspects such as their morphometric, behavioral, and physiologic characteristics, mtDNA and genetic background [26], and immunologic parameters and immune response to infections [27][28][29], differences in their MHC genotypes might also exist.…”

A Rapid Progressor-Specific Variant Clone of Simian Immunodeficiency Virus Replicates Efficiently In Vivo Only in the Absence of Immune Reponses

“…The length of CD8 ϩ T-cell depletion is also critical, since disease is most rapid in animals with long-term depletion (59). The rate of disease progression in HIV/SIV infection is also influenced by major histocompatibility complex alleles (4,12,27,58). Additionally, the intrinsic susceptibility of CD4…”

Increased reproductive success of MHC class II heterozygous males among free-ranging rhesus macaques

“…Mamu-DQB typing was performed as previously described (Sauermann et al 1996). Rhesus DQA and DQB loci are in strong linkage disequilibrium (Sauermann 1998), so that homozygosity for DQB1 could be confirmed by DQA1 analysis in order to minimize the risk of typing artefacts.…”