Australian Journal of Zoology Australian Journal of Zoology Society
Evolutionary, molecular and comparative zoology

Hatching time for monotreme immunology

Emily S. W. Wong A , Anthony T. Papenfuss B , Robert D. Miller C and Katherine Belov A D

A Faculty of Veterinary Science, B19 RMC Gunn, University of Sydney, Sydney, NSW 2006, Australia.

B Bioinformatics Division, The Walter and Eliza Hall Institute for Medical Research, Parkville, Vic. 3052, Australia.

C Center for Evolutionary and Theoretical Immunology, Department of Biology, University of New Mexico, Albuquerque, NM, USA.

D Corresponding author. Email:

Australian Journal of Zoology 57(4) 185-198
Submitted: 7 April 2009  Accepted: 6 July 2009   Published: 26 October 2009


The sequencing of the platypus genome has spurred investigations into the characterisation of the monotreme immune response. As the most divergent of extant mammals, the characterisation of the monotreme immune repertoire allows us to trace the evolutionary history of immunity in mammals and provide insights into the immune gene complement of ancestral mammals. The immune system of monotremes has remained largely uncharacterised due to the lack of specific immunological reagents and limited access to animals for experimentation. Early immunological studies focussed on the anatomy and physiology of the lymphoid system in the platypus. More recent molecular studies have focussed on characterisation of individual immunoglobulin, T-cell receptor and MHC genes in both the platypus and short-beaked echidna. Here, we review the published literature on the monotreme immune gene repertoire and provide new data generated from genome analysis on cytokines, Fc receptors and immunoglobulins. We present an overview of key gene families responsible for innate and adaptive immunity including the cathelicidins, defensins, T-cell receptors and the major histocompatibility complex (MHC) Class I and Class II antigens. We comment on the usefulness of these sequences for future studies into immunity, health and disease in monotremes.


Altschul S. F. Madden T. L. Schaffer A. A. Zhang J. Zhang Z. Miller W. Lipman D. J. 1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25 3389 3402 doi:10.1093/nar/25.17.3389

Beck G. 1998 Macrokines: invertebrate cytokine-like molecules? Frontiers in Bioscience: A Journal and Virtual Library 3 d559 d569

Belov K. Hellman L. 2003 a Immunoglobulin genetics of Ornithorhynchus anatinus (platypus) and Tachyglossus aculeatus (short-beaked echidna). Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 13 811 819 doi:10.1016/S1095-6433(03)00165-X

Belov K. Hellman L. 2003 b Platypus immunoglobulin M and the divergence of the two extant monotreme lineages. Australian Mammalogy 25 87 94

Belov K. Hellman L. Cooper D. W. 2002 a Characterization of immunoglobulin gamma 1 from a monotreme, Tachyglossus aculeatus. Immunogenetics 53 1065 1071 doi:10.1007/s00251-002-0432-1

Belov K. Zenger K. R. Hellman L. Cooper D. W. 2002 b Echidna IgA supports mammalian unity and traditional Therian relationship. Mammalian Genome 13 656 663 doi:10.1007/s00335-002-3004-7

Belov K. Lam M. K. Hellman L. Colgan D. J. 2003 Evolution of the major histocompatibility complex: Isolation of class II beta cDNAs from two monotremes, the platypus and the short-beaked echidna. Immunogenetics 55 402 411 doi:10.1007/s00251-003-0598-1

Belov K. Miller R. D. Ilijeski A. Hellman L. Harrison G. A. 2004 Isolation of monotreme T-cell receptor alpha and beta chains. Immunogenetics 56 164 169 doi:10.1007/s00251-004-0679-9

Belov K. Deakin J. E. Papenfuss A. T. Baker M. L. Melman S. D. Siddle H. V. Gouin N. Goode D. L. Sargeant T. J. Robinson M. D. Wakefield M. J. Mahony S. Cross J. G. R. Benos P. V. Samollow P. B. Speed T. P. Graves J. A. M. Miller R. D. 2006 Reconstructing an ancestral mammalian immune supercomplex from a marsupial major histocompatibility complex. PLoS Biology 4 3 e46

Belov K. Sanderson C. E. Deakin J. E. Wong E. S. Assange D., et al 2007 Characterization of the opossum immune genome provides insights into the evolution of the mammalian immune system. Genome Research 17 982 991 doi:10.1101/gr.6121807

Bininda-Emonds O. R. Cardillo M. Jones K. E. MacPhee R. D. Beck R. M. Grenyer R. Price S. A. Vos R. A. Gittleman J. L. Purvis A. 2007 The delayed rise of present-day mammals. Nature 446 507 512 doi:10.1038/nature05634

Brown K. L. Hancock R. E. 2006 Cationic host defense (antimicrobial) peptides. Current Opinion in Immunology 18 24 30 doi:10.1016/j.coi.2005.11.004

Connolly J. H. Canfield P. J. McClure S. J. Whittington R. J. 1999 Histological and immunohistological investigation of lymphoid tissue in the platypus (Ornithorhynchus anatinus). Journal of Anatomy 195 161 171 doi:10.1046/j.1469-7580.1999.19520161.x

Daly K. A. Digby M. R. Lefèvre C. Nicholas K. R. Deane E. M. Williamson P. 2008 Identification, characterization and expression of cathelicidin in the pouch young of tammar wallaby (Macropus eugenii). Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology 149 524 533 doi:10.1016/j.cbpb.2007.12.002

Davis R. S. 2007 Fc receptor-like molecules. Annual Review of Immunology 25 525 560 doi:10.1146/annurev.immunol.25.022106.141541

Diener E. Ealey E. H. M. 1965 Immune system in a monotreme: studies on the Australian echidna (Tachyglossus aculeatus). Nature 208 950 953 doi:10.1038/208950a0

Diener E. Wister R. Ealey E. H. M. 1967 Phylogenetic studies on the immune response II. The immune response of the Australian echidna Tachyglossus aculeatus. Immunology 13 329 337

Dohm J. C. Tsend-Ayush E. Reinhardt R. Grützner F. Himmelbauer H. 2007 Disruption and pseudoautosomal localization of the major histocompatibility complex in monotremes. Genome Biology 8 R175 doi:10.1186/gb-2007-8-8-r175

Eddy S. R. 1998 Profile hidden Markov models. Bioinformatics (Oxford, England) 14 755 763 doi:10.1093/bioinformatics/14.9.755

Fayngerts S. A. Najakshin A. M. Taranin A. V. 2007 Species-specific evolution of the FcR family in endothermic vertebrates. Immunogenetics 59 493 506 doi:10.1007/s00251-007-0208-8

Flajnik M. F. Ohta Y. Greenberg A. S. Salter-Cid L. Carrizosa A. Du Pasquier L. Kasahara M. 1999 Two ancient allelic lineages at the single classical class I locus in the Xenopus MHC. Journal of Immunology 163 3826 3833

Fling S. P. Arp B. Pious D. 1994 HLA-DMA and -DMB genes are both required for MHC class II/peptide complex formation in antigen-presenting cells. Nature 368 554 558 doi:10.1038/368554a0

Freyer C. Renfree M. B. 2008 The mammalian yolk sac placenta. Journal of Experimental Zoology. Part B. Molecular and Developmental Evolution 310B 1 10

Guselnikov S. V. Ramanayake T. Erilova A. Y. Mechetina L. V. Najakshin A. M. Robert J. Taranin A. V. 2008 The Xenopus FcR family demonstrates continually high diversification of paired receptors in vertebrate evolution. BMC Evolutionary Biology 8 148 doi:10.1186/1471-2148-8-148

Harrison G. A. McNicol K. A. Deane E. M. 2004 Type I interferon genes from the egg-laying mammal, Tachyglossus aculeatus (short-beaked echidna). Immunology and Cell Biology 82 112 118 doi:10.1046/j.0818-9641.2004.01230.x

Haynes J. I. 2001 The marsupial and monotreme thymus, revisited. Journal of Zoology 253 167 173 doi:10.1017/S0952836901000152

Horton R. Wilming L. Rand V. Lovering R. C. Bruford E. A., et al 2004 Gene map of the extended human MHC. Nature Reviews. Genetics 5 889 899 doi:10.1038/nrg1489

Huttner K. M. Lambeth M. R. Burkin H. R. Burkin D. J. Broad T. E. 1998 Localization and genomic organization of sheep antimicrobial peptide genes. Gene 206 85 91 doi:10.1016/S0378-1119(97)00569-6

Janeway C. A. , Travers P. , Walport M. , and Shlomchik M. (2001). ‘Immunobiology.’ 5th edn. (Garland Publishing: New York.)

Johansson J. Aveskogh M. Munday B. Hellman L. 2002 Heavy chain V region diversity in the duck-billed platypus (Ornithorhynchus anatinus): long and highly variable complementarity-determining region 3 compensates for limited germline diversity. Journal of Immunology 168 5155 5162

Jones D. T. Taylor W. R. Thornton J. M. 1992 The rapid generation of mutation data matrices from protein sequences. Computer Applications in the Biosciences 8 275 282

Kaplan N. Morpurgo N. Linial M. 2007 Novel families of toxin-like peptides in insects and mammals: a computational approach. Journal of Molecular Biology 369 553 566 doi:10.1016/j.jmb.2007.02.106

Kaufman J. Milne S. Gobel T. W. F. Walker B. A. Jacob J. P. Auffray C. Zoorob R. Beck S. 1999 The chicken B locus is a minimal essential major histocompatibility complex. Nature 401 923 925 doi:10.1038/44856

Kordis D. Lovsin N. Gubensek F. 2006 Phylogenomic analysis of the L1 retrotransposons in Deuterostomia. Systematic Biology 55 886 901 doi:10.1080/10635150601052637

Kumanovics A. Takada T. Lindahl K. F. 2003 Genomic organization of the mammalian MHC. Annual Review of Immunology 21 629 657 doi:10.1146/annurev.immunol.21.090501.080116

Lefèvre C. Digby M. Whitley J. Strahm Y. Nicholas K. 2007 Lactation transcriptomics in the Australian marsupial, Macropus eugenii: transcript sequencing and quantification. BMC Genomics 8 417 doi:10.1186/1471-2164-8-417

Maruoka T. Nagata T. Kasahara M. 2004 Identification of the rat IgA Fc receptor encoded in the leukocyte receptor complex. Immunogenetics 55 712 716 doi:10.1007/s00251-003-0626-1

Miska K. B. Harrison G. A. Hellman L. Miller R. D. 2002 The major histocompatibility complex in monotremes: an analysis of the evolution of Mhc class I genes across all three mammalian subclasses. Immunogenetics 54 381 393 doi:10.1007/s00251-002-0484-2

Miska K. B. Hellman L. Miller R. D. 2003 Characterization of beta(2)-microglobulin coding sequence from three non-placental mammals: the duckbill platypus, the short-beaked echidna, and the grey short-tailed opossum. Developmental and Comparative Immunology 27 247 256 doi:10.1016/S0145-305X(02)00095-2

Old J. M. Deane E. M. 2000 Development of the immune system and immunological protection in marsupial pouch young. Developmental and Comparative Immunology 24 445 454 doi:10.1016/S0145-305X(00)00008-2

Parra Z. E. Arnold T. Nowak M. A. Hellman L. Miller R. D. 2006 TCR gamma chain diversity in the spleen of the duckbill platypus (Ornithorhynchus anatinus). Developmental and Comparative Immunology 30 699 710 doi:10.1016/j.dci.2005.10.002

Parra Z. E. Baker M. L. Schwarz R. S. Deakin J. E. Lindblad-Toh K. Miller R. D. 2007 A unique T cell receptor discovered in marsupials. Proceedings of the National Academy of Sciences of the United States of America 104 9776 9781 doi:10.1073/pnas.0609106104

Parra Z. E. Baker M. L. Hathaway J. Lopez A. M. Trujillo J. Sharp A. Miller R. D. 2008 Comparative genomic analysis and evolution of the T cell receptor loci in the opossum Monodelphis domestica. BMC Genomics 9 111 doi:10.1186/1471-2164-9-111

Ramanathan B. Davis E. G. Ross C. R. Blecha F. 2002 Cathelicidins: microbicidal activity, mechanisms of action, and roles in innate immunity. Microbes and Infection 4 361 372 doi:10.1016/S1286-4579(02)01549-6

Rens W. Grützner F. O’Brien P. C. M. Fairclough H. Graves J. A. M. Ferguson-Smith M. A. 2004 Resolution and evolution of the duck-billed platypus karyotype with an X1Y1X2Y2X3Y3X4Y4X5Y5 male sex chromosome constitution. Proceedings of the National Academy of Sciences of the United States of America 101 16257 16261 doi:10.1073/pnas.0405702101

Rowlands D. T. 1970 The immune response of adult opossums (Didelphis virginiana) to the bacteriophage f2. Immunology 18 149 155

Scocchi M. Wang S. Zanetti M. 1997 Structural organization of the bovine cathelicidin gene family and identification of a novel member. FEBS Letters 417 311 315 doi:10.1016/S0014-5793(97)01310-0

Scocchi M. Bontempo D. Boscolo S. Tomasinsig L. Giulotto E. Zanetti M. 1999 Novel cathelicidins in horse leukocytes(1). FEBS Letters 457 459 464 doi:10.1016/S0014-5793(99)01097-2

Shibuya A. Sakamoto N. Shimizu Y. Shibuya K. Osawa M., et al 2000 Fc alpha/mu receptor mediates endocytosis of IgM-coated microbes. Nature Immunology 1 441 446 doi:10.1038/80886

Shimizu Y. Honda S. Yotsumoto K. Tahara-Hanaoka S. Eyre H. J., et al 2001 Fc(alpha)/mu receptor is a single gene-family member closely related to polymeric immunoglobulin receptor encoded on Chromosome 1. Immunogenetics 53 709 711 doi:10.1007/s00251-001-0375-y

Skerlavaj B. Scocchi M. Gennaro R. Risso A. Zanetti M. 2001 Structural and functional analysis of horse cathelicidin peptides. Antimicrobial Agents and Chemotherapy 45 715 722 doi:10.1128/AAC.45.3.715-722.2001

Stone W. H. Brunn D. A. Foster E. B. Manis G. S. Hoffman E. S. Saphire D. G. VandeBerg J. L. Infante A. J. 1998 Absence of a significant mixed lymphocyte reaction in a marsupial (Monodelphis domestica). Laboratory Animal Science 48 184 189

Takahashi K. Rooney A. P. Nei M. 2000 Origins and divergence times of mammalian class II MHC gene clusters. The Journal of Heredity 91 198 204 doi:10.1093/jhered/91.3.198

Tamura K. Dudley J. Nei M. Kumar S. 2007 MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24 1596 1599 doi:10.1093/molbev/msm092

Taylor A. I. Gould H. J. Sutton B. J. Calvert R. A. 2007 The first avian Ig-like Fc receptor family member combines features of mammalian FcR and FCRL. Immunogenetics 59 323 328 doi:10.1007/s00251-007-0195-9

Temple-Smith P. Grant T. 2001 Uncertain breeding: a short history of reproduction in monotremes. Reproduction, Fertility and Development 13 487 497 doi:10.1071/RD01110

Trowsdale J. 1995 “Both man & bird & beast”: comparative organization of MHC genes. Immunogenetics 41 1 17 doi:10.1007/BF00188427

Tyndale-Biscoe C. H. , and Renfree M. (1987). ‘Reproductive Physiology of Marsupials.’(Cambridge University Press: Cambridge, UK.)

Vernersson M. Aveskogh M. Munday B. Hellman L. 2002 Evidence for an early appearance of modern post-switch immunoglobulin isotypes in mammalian evolution (II); cloning of IgE, IgG1 and IgG2 from a monotreme, the duck-billed platypus, Ornithorhynchus anatinus. European Journal of Immunology 32 2145 2155 doi:10.1002/1521-4141(200208)32:8<2145::AID-IMMU2145>3.0.CO;2-I

Warren W. C. Hillier L. W. Graves J. A. M. Birney E. Ponting C. P., et al 2008 Genome analysis of the platypus reveals unique signatures of evolution. Nature 453 175 183 doi:10.1038/nature06936

Whittington C. M. Papenfuss A. T. Bansal P. Torres A. M. Wong E. S., et al 2008 Defensins and the convergent evolution of platypus and reptile venom genes. Genome Research 18 986 994 doi:10.1101/gr.7149808

Wieland W. H. Orzaez D. Lammers A. Parmentier H. K. Verstegen M. W. Schots A. 2004 A functional polymeric immunoglobulin receptor in chicken (Gallus gallus) indicates ancient role of secretory IgA in mucosal immunity. The Biochemical Journal 380 669 676 doi:10.1042/BJ20040200

Wong E. Young L. Papenfuss A. Belov K. 2006 In silico identification of opossum cytokine genes suggests the complexity of the marsupial immune system rivals that of eutherian mammals. Immunome Research 2 4 doi:10.1186/1745-7580-2-4

Wronski E. V. Woods G. M. Munday B. L. 2003 Antibody response to sheep red blood cells in platypus and echidna. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 136 957 963 doi:10.1016/S1095-6433(03)00325-8

Yeh R. F. Lim L. P. Burge C. B. 2001 Computational inference of homologous gene structures in the human genome. Genome Research 11 803 816 doi:10.1101/gr.175701

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