, Volume 62, Issue 3, pp 123–136 | Cite as

Study of cynomolgus monkey (Macaca fascicularis) DRA polymorphism in four populations

  • Alice Aarnink
  • Laurent Estrade
  • Pol-André Apoil
  • Yuki F. Kita
  • Naruya Saitou
  • Takashi Shiina
  • Antoine Blancher
Original Paper


To describe the polymorphism of the DRA gene in Macaca fascicularis, we have studied 141 animals either at cDNA level (78 animals from Mauritius, the Philippines, and Vietnam) or genomic level (63 animals from the Philippines, Indonesia, and Vietnam). In total, we characterized 22 cDNA DRA alleles, 13 of which had not been described until now. In the Mauritius population, we confirmed the presence of three DRA alleles. In the Philippine and Vietnam populations, we observed 11 and 14 DRA alleles, respectively. Only two alleles were present in all three populations. All DRA alleles but one differ from the consensus sequence by one to three mutations, most being synonymous; so, only seven DR alpha proteins were deduced from the 22 cDNA alleles. One DRA cDNA allele, Mafa-DRA*02010101, differs from all other alleles by 11 to 14 mutations of which only four are non-synonymous. The two amino acid changes inside the peptide groove of Mafa-DRA*02010101 are highly conservative. The very low proportion of non-synonymous/synonymous mutations is compatible with a purifying selection which is comparable to all previous observations concerning the evolution of the DRA gene in mammals. Homologues of the allele Mafa-DRA*02010101 are also found in two other Asian macaques (Macaca mulatta and Macaca nemestrina). The forces able to maintain this highly divergent allele in three different macaque species remain hypothetical.


MHC DRA Macaque Polymorphism 



This work was performed with funds from the French Ministry of Research (EA3034 contract), from a research grant from Novartis, and with the help of ANRS (contract number 07375). Other funds were from the Laboratory of Dr. Takashi Shiina. We thank Pr. Brigitte Le Mauf (Nantes, France), Dominic Borie (Stanford, USA), Eric André (Bioprim, Baziège, France), Marc Bigaud (Novartis, Bâle), Dr. Jacques Ducos de Lahitte (Ecole Nationale Vétérinaire de Toulouse), for their help in obtaining macaque blood samples. Our thanks are also due to Peter Winterton for his contribution in drawing up the final version of this manuscript.

We are pleased to thank all the technicians of the Toulouse Laboratory of Immunogenetics: Béatrice Atlan, Audrey Dauba, Stéphanie Despiau-Schiavinato, and Sylvie Hébrard for their excellent technical assistance.

Supplementary material

251_2009_421_MOESM1_ESM.doc (252 kb)
ESM 1 (DOC 267 kb)


  1. Aoyama A, Ng CY, Millington TM, Boskovic S, Murakami T, Wain JC, Houser SL, Madsen JC, Kawai T, Allan JS (2009) Comparison of lung and kidney allografts in induction of tolerance by a mixed-chimerism approach in cynomolgus monkeys. Transplant Proc 41:429–430CrossRefPubMedGoogle Scholar
  2. Baskin CR, Bielefeldt-Ohmann H, Tumpey TM, Sabourin PJ, Long JP, Garcia-Sastre A, Tolnay AE, Albrecht R, Pyles JA, Olson PH, Aicher LD, Rosenzweig ER, Murali-Krishna K, Clark EA, Kotur MS, Fornek JL, Proll S, Palermo RE, Sabourin CL, Katze MG (2009) Early and sustained innate immune response defines pathology and death in nonhuman primates infected by highly pathogenic influenza virus. Proc Natl Acad Sci USA 106:3455–3460CrossRefPubMedGoogle Scholar
  3. Belkhir K, Borsa P, Chikhi L, Raufaste N, Bonhomme F (2004) GENETIX 4.05, a software for population genetics.
  4. Bimber BN, Moreland AJ, Wiseman RW, Hughes AL, O'Connor DH (2008) Complete characterization of killer Ig-like receptor (KIR) haplotypes in Mauritian cynomolgus macaques: novel insights into nonhuman primate KIR gene content and organization. J Immunol 181:6301–6308PubMedGoogle Scholar
  5. Blancher A, Tisseyre P, Dutaur M, Apoil PA, Maurer C, Quesniaux V, Raulf F, Bigaud M, Abbal M (2006) Study of cynomolgus monkey (Macaca fascicularis) MhcDRB (Mafa-DRB) polymorphism in two populations. Immunogenetics 58:269–282CrossRefPubMedGoogle Scholar
  6. Blancher A, Bonhomme M, Crouau-Roy B, Terao K, Kitano T, Saitou N (2008) Mitochondrial DNA sequence phylogeny of 4 populations of the widely distributed cynomolgus macaque (Macaca fascicularis fascicularis). J Hered 99:254–264CrossRefPubMedGoogle Scholar
  7. Bonhomme M, Blancher A, Crouau-Roy B (2005) Multiplexed microsatellites for rapid identification and characterization of individuals and populations of Cercopithecidae. Am J Primatol 67:385–391CrossRefPubMedGoogle Scholar
  8. Bonhomme M, Blancher A, Jalil MF, Crouau-Roy B (2007) Factors shaping genetic variation in the MHC of natural non-human primate populations. Tissue Antigens 70:398–411CrossRefPubMedGoogle Scholar
  9. Bonhomme M, Blancher A, Cuartero S, Chikhi L, Crouau-Roy B (2008) Origin and number of founders in an introduced insular primate: estimation from nuclear genetic data. Mol Ecol 17:1009–1019CrossRefPubMedGoogle Scholar
  10. Bonhomme M, Cuartero S, Blancher A, Crouau-Roy B (2009) Assessing natural introgression in 2 biomedical model species, the rhesus macaque (Macaca mulatta) and the long-tailed macaque (Macaca fascicularis). J Hered 100:158–169CrossRefPubMedGoogle Scholar
  11. Borie D, Hausen B, Larson M, Klupp J, Stalder M, Birsan T, Morris R (2002) A life-supporting technique of renal allotransplantation in Macaca fascicularis to evaluate novel immunosuppressive drugs in nonhuman primates. J Surg Res 107:64–74PubMedGoogle Scholar
  12. Burwitz BJ, Pendley CJ, Greene JM, Detmer AM, Lhost JJ, Karl JA, Piaskowski SM, Rudersdorf RA, Wallace LT, Bimber BN, Loffredo JT, Cox DG, Bardet W, Hildebrand W, Wiseman RW, O'Connor SL, O'Connor DH (2009) Mauritian cynomolgus macaques share two exceptionally common major histocompatibility complex class I alleles that restrict simian immunodeficiency virus-specific CD8+ T cells. J Virol 83:6011–6019CrossRefPubMedGoogle Scholar
  13. Calvas P, Apoil P, Fortenfant F, Roubinet F, Andris J, Capra D, Blancher A (1999) Characterization of the three immunoglobulin G subclasses of macaques. Scand J Immunol 49:595–610CrossRefPubMedGoogle Scholar
  14. Campbell KJ, Detmer AM, Karl JA, Wiseman RW, Blasky AJ, Hughes AL, Bimber BN, O'Connor SL, O'Connor DH (2009) Characterization of 47 MHC class I sequences in Filipino cynomolgus macaques. Immunogenetics 61:177–187CrossRefPubMedGoogle Scholar
  15. Daza-Vamenta R, Glusman G, Rowen L, Guthrie B, Geraghty DE (2004) Genetic divergence of the rhesus macaque major histocompatibility complex. Genome Res 14:1501–1515CrossRefPubMedGoogle Scholar
  16. de Groot N, Doxiadis GG, De Groot NG, Otting N, Heijmans C, Rouweler AJ, Bontrop RE (2004) Genetic makeup of the DR region in rhesus macaques: gene content, transcripts, and pseudogenes. J Immunol 172:6152–6157PubMedGoogle Scholar
  17. Derdouch S, Gay W, Negre D, Prost S, Le Dantec M, Delache B, Auregan G, Andrieu T, Leplat JJ, Cosset FL, Le Grand R (2008) Reconstitution of the myeloid and lymphoid compartments after the transplantation of autologous and genetically modified CD34+ bone marrow cells, following gamma irradiation in cynomolgus macaques. Retrovirology 5:50CrossRefPubMedGoogle Scholar
  18. Doxiadis GG, de Groot N, de Groot NG, Doxiadis II, Bontrop RE (2008) Reshuffling of ancient peptide binding motifs between HLA-DRB multigene family members: old wine served in new skins. Mol Immunol 45:2743–2751CrossRefPubMedGoogle Scholar
  19. Excoffier L, Laval G, Schneider S (2005) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evol Bioinform Online 1:47–50PubMedGoogle Scholar
  20. Florese RH, Wiseman RW, Venzon D, Karl JA, Demberg T, Larsen K, Flanary L, Kalyanaraman VS, Pal R, Titti F, Patterson LJ, Heath MJ, O'Connor DH, Cafaro A, Ensoli B, Robert-Guroff M (2008) Comparative study of Tat vaccine regimens in Mauritian cynomolgus and Indian rhesus macaques: influence of Mauritian MHC haplotypes on susceptibility/resistance to SHIV (89.6P) infection. Vaccine 26:3312–3321CrossRefPubMedGoogle Scholar
  21. Hughes AL, Nei M (1989) Nucleotide substitution at major histocompatibility complex class II loci: evidence for overdominant selection. Proc Natl Acad Sci USA 86:958–962CrossRefPubMedGoogle Scholar
  22. Kita Y, Tanaka T, Yoshida S, Ohara N, Kaneda Y, Kuwayama S, Muraki Y, Kanamaru N, Hashimoto S, Takai H, Okada C, Fukunaga Y, Sakaguchi Y, Furukawa I, Yamada K, Inoue Y, Takemoto Y, Naito M, Yamada T, Matsumoto M, McMurray DN, Cruz EC, Tan EV, Abalos RM, Burgos JA, Gelber R, Skeiky Y, Reed S, Sakatani M, Okada M (2005) Novel recombinant BCG and DNA-vaccination against tuberculosis in a cynomolgus monkey model. Vaccine 23:2132–2135CrossRefPubMedGoogle Scholar
  23. Klein J (1987) Origin of major histocompatibility complex polymorphism: the trans-species hypothesis. Hum Immunol 19:155–162CrossRefPubMedGoogle Scholar
  24. Klein J, Bontrop RE, Dawkins RL, Erlich HA, Gyllensten UB, Heise ER, Jones PP, Parham P, Wakeland EK, Watkins DI (1990) Nomenclature for the major histocompatibility complexes of different species: a proposal. Immunogenetics 31:217–219PubMedGoogle Scholar
  25. Kuiken T, Rimmelzwaan GF, Van Amerongen G, Osterhaus AD (2003) Pathology of human influenza A (H5N1) virus infection in cynomolgus macaques (Macaca fascicularis). Vet Pathol 40:304–310CrossRefPubMedGoogle Scholar
  26. Lau M, Vayntrub T, Grumet FC, Lowsky R, Strober S, Hoppe R, Larson M, Holm B, Reitz B, Borie D (2004) Short tandem repeat analysis to monitor chimerism in Macaca fascicularis. Am J Transplant 4:1543–1548CrossRefPubMedGoogle Scholar
  27. Lawler SH, Sussman RW, Taylor LL (1995) Mitochondrial DNA of the Mauritian macaques (Macaca fascicularis): an example of the founder effect. Am J Phys Anthropol 96:133–141CrossRefPubMedGoogle Scholar
  28. Lawler JV, Endy TP, Hensley LE, Garrison A, Fritz EA, Lesar M, Baric RS, Kulesh DA, Norwood DA, Wasieloski LP, Ulrich MP, Slezak TR, Vitalis E, Huggins JW, Jahrling PB, Paragas J (2006) Cynomolgus macaque as an animal model for severe acute respiratory syndrome. PLoS Med 3:e149CrossRefPubMedGoogle Scholar
  29. Leuchte N, Berry N, Kohler B, Almond N, LeGrand R, Thorstensson R, Titti F, Sauermann U (2004) MhcDRB-sequences from cynomolgus macaques (Macaca fascicularis) of different origin. Tissue Antigens 63:529–537CrossRefPubMedGoogle Scholar
  30. Mee ET, Badhan A, Karl JA, Wiseman RW, Cutler K, Knapp LA, Almond N, O'Connor DH, Rose NJ (2009a) MHC haplotype frequencies in a UK breeding colony of Mauritian cynomolgus macaques mirror those found in a distinct population from the same geographic origin. J Med Primatol 38:1–14CrossRefPubMedGoogle Scholar
  31. Mee ET, Berry N, Ham C, Sauermann U, Maggiorella MT, Martinon F, Verschoor EJ, Heeney JL, Le Grand R, Titti F, Almond N, Rose NJ (2009b) Mhc haplotype H6 is associated with sustained control of SIVmac251 infection in Mauritian cynomolgus macaques. Immunogenetics 61:327–339CrossRefPubMedGoogle Scholar
  32. Menninger K, Wieczorek G, Riesen S, Kunkler A, Audet M, Blancher A, Schuurman HJ, Quesniaux V, Bigaud M (2002) The origin of cynomolgus monkey affects the outcome of kidney allografts under Neoral immunosuppression. Transplant Proc 34:2887–2888CrossRefPubMedGoogle Scholar
  33. Morner A, Douagi I, Forsell MN, Sundling C, Dosenovic P, O'Dell S, Dey B, Kwong PD, Voss G, Thorstensson R, Mascola JR, Wyatt RT, Karlsson Hedestam GB (2009) Human immunodeficiency virus type 1 env trimer immunization of macaques and impact of priming with viral vector or stabilized core protein. J Virol 83:540–551CrossRefPubMedGoogle Scholar
  34. O'Connor SL, Blasky AJ, Pendley CJ, Becker EA, Wiseman RW, Karl JA, Hughes AL, O'Connor DH (2007) Comprehensive characterization of MHC class II haplotypes in Mauritian cynomolgus macaques. Immunogenetics 59:449–462CrossRefPubMedGoogle Scholar
  35. Otting N, de Groot NG, Doxiadis GG, Bontrop RE (2002) Extensive Mhc-DQB variation in humans and non-human primate species. Immunogenetics 54:230–239CrossRefPubMedGoogle Scholar
  36. Osada N, Hirata M, Tanuma R, Suzuki Y, Sugano S, Terao K, Kusuda J, Kameoka Y, Hashimoto K, Takahashi I (2009) Collection of Macaca fascicularis cDNAs derived from bone marrow, kidney, liver, pancreas, spleen, and thymus. BMC Res Notes 2:199CrossRefPubMedGoogle Scholar
  37. Pendley CJ, Becker EA, Karl JA, Blasky AJ, Wiseman RW, Hughes AL, O'Connor SL, O'Connor DH (2008) MHC class I characterization of Indonesian cynomolgus macaques. Immunogenetics 60:339–351CrossRefPubMedGoogle Scholar
  38. Puissant B, Abbal M, Blancher A (2003) Polymorphism of human and primate RANTES, CX3CR1, CCR2 and CXCR4 genes with regard to HIV/SIV infection. Immunogenetics 55:275–283CrossRefPubMedGoogle Scholar
  39. Reche P, Reinherz E (2003) Sequence variability analysis of human class I and class II MHC molecules: functional and structural correlates of amino acid polymorphisms. J Mol Biol 331:623–641CrossRefPubMedGoogle Scholar
  40. Reed DS, Larsen T, Sullivan LJ, Lind CM, Lackemeyer MG, Pratt WD, Parker MD (2005) Aerosol exposure to western equine encephalitis virus causes fever and encephalitis in cynomolgus macaques. J Infect Dis 192:1173–1182CrossRefPubMedGoogle Scholar
  41. Robinson J, Waller MJ, Stoehr P, Marsh SG (2005) IPD—the Immuno Polymorphism Database. Nucleic Acids Res 33:D523–D526CrossRefPubMedGoogle Scholar
  42. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  43. Sano K, Shiina T, Kohara S, Yanagiya K, Hosomichi K, Shimizu S, Anzai T, Watanabe A, Ogasawara K, Torii R, Kulski JK, Inoko H (2006) Novel cynomolgus macaque MHC-DPB1 polymorphisms in three South-East Asian populations. Tissue Antigens 67:297–306CrossRefPubMedGoogle Scholar
  44. Sato H, Kobune F, Ami Y, Yoneda M, Kai C (2008) Immune responses against measles virus in cynomolgus monkeys. Comp Immunol Microbiol Infect Dis 31:25–35CrossRefPubMedGoogle Scholar
  45. Satta Y (1993) Balancing selection at HLA loci. In: Takahata N, Clark AG (eds) Mechanism of molecular evolution. Japan Scientific Societies Press Sinauer Associated Inc., Tokyo, JapanGoogle Scholar
  46. Schmidt LH, Fradkin R, Harrison J, Rossan RN (1977) Differences in the virulence of Plasmodium knowlesi for Macaca irus (fascicularis) of Philippine and Malayan origins. Am J Trop Med Hyg 26:612–622PubMedGoogle Scholar
  47. Street SL, Kyes RC, Grant R, Ferguson B (2007) Single nucleotide polymorphisms (SNPs) are highly conserved in rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) macaques. BMC Genomics 8:480CrossRefPubMedGoogle Scholar
  48. Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 101:11030–11035CrossRefPubMedGoogle Scholar
  49. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599CrossRefPubMedGoogle Scholar
  50. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680CrossRefPubMedGoogle Scholar
  51. Tosi AJ, Coke CS (2007) Comparative phylogenetics offer new insights into the biogeographic history of Macaca fascicularis and the origin of the Mauritian macaques. Mol Phylogenet Evol 42:498–504CrossRefPubMedGoogle Scholar
  52. Turbant S, Martinon F, Moine G, Le Grand R, Leonetti M (2009) Cynomolgus macaques immunized with two HIV-1 Tat stabilized proteins raise strong and long-lasting immune responses with a pattern of Th1/Th2 response differing from that in mice. Vaccine 27:5349–5356CrossRefPubMedGoogle Scholar
  53. Uda A, Tanabayashi K, Yamada YK, Akari H, Lee YJ, Mukai R, Terao K, Yamada A (2004) Detection of 14 alleles derived from the MHC class I A locus in cynomolgus monkeys. Immunogenetics 56:155–163CrossRefPubMedGoogle Scholar
  54. Uda A, Tanabayashi K, Fujita O, Hotta A, Terao K, Yamada A (2005) Identification of the MHC class I B locus in cynomolgus monkeys. Immunogenetics 57:189–197CrossRefPubMedGoogle Scholar
  55. Voorter CBR, Van den Berg-Loonen E (2006) The HLADRA gene and its extending polymorphism (abstract, 32nd Annual ASHI Meeting). Human Immunology 67:S106CrossRefGoogle Scholar
  56. Warfield KL, Swenson DL, Olinger GG, Kalina WV, Aman MJ, Bavari S (2007) Ebola virus-like particle-based vaccine protects nonhuman primates against lethal Ebola virus challenge. J Infect Dis 196(Suppl 2):S430–S437CrossRefPubMedGoogle Scholar
  57. Watanabe A, Shiina T, Shimizu S, Hosomichi K, Yanagiya K, Kita YF, Kimura T, Soeda E, Torii R, Ogasawara K, Kulski JK, Inoko H (2007) A BAC-based contig map of the cynomolgus macaque (Macaca fascicularis) major histocompatibility complex genomic region. Genomics 89:402–412CrossRefPubMedGoogle Scholar
  58. Wieczorek G, Bigaud M, Menninger K, Riesen S, Quesniaux V, Schuurman HJ, Audet M, Blancher A, Mihatsch MJ, Nickeleit V (2006) Acute and chronic vascular rejection in nonhuman primate kidney transplantation. Am J Transplant 6:1285–1296CrossRefPubMedGoogle Scholar
  59. Wiseman RW, Wojcechowskyj JA, Greene JM, Blasky AJ, Gopon T, Soma T, Friedrich TC, O'Connor SL, O'Connor DH (2007) Simian immunodeficiency virus SIVmac239 infection of major histocompatibility complex-identical cynomolgus macaques from Mauritius. J Virol 81:349–361CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Alice Aarnink
    • 1
  • Laurent Estrade
    • 1
  • Pol-André Apoil
    • 1
  • Yuki F. Kita
    • 2
  • Naruya Saitou
    • 3
  • Takashi Shiina
    • 2
  • Antoine Blancher
    • 1
  1. 1.Laboratoire d’immunogénétique moléculaire, EA3034, Faculté de Médecine Purpan, Toulouse 3, IFR150 (INSERM), CHU de ToulouseUniversité Paul SabatierToulouse Cedex 9France
  2. 2.Department of Molecular Life Science, Division of Basic Medical Science and Molecular MedicineTokai University School of MedicineIseharaJapan
  3. 3.Division of Population GeneticsNational Institute of GeneticsMishimaJapan

Personalised recommendations