The Pigtail Macaque (Macaca nemestrina) in Biomedical Research

Charlotte E. Hotchkiss The Macaca nemestrina (pigtail macaques) is an invaluable non-human primate model that has been utilized to support research in the areas of reproduction and development, immunology, infectious disease, neuroscience, transplantation and stem cell research.1-30 This summary outlines the natural history of the M. nemestrina and summarizes the important research areas that have benefited from having this unique animal model available for advancing biomedical research. Natural Biology The southern pigtail macaque (M. nemestrina) is native to southeast Asia, primarily inhabiting islands in Indonesia, but also the mainland in Malaysia, Thailand, and Brunei Darussalam.31 It is considered a separate species from the northern pigtail macaque (M. leonina) based on morphological characteristics,32 but there is range overlap, and interbreeding occurs in Thailand and on the islands of Phuket and Yao Yai.33 Genetically, M. nemestrina is in the silenus group34 which is distinct from the macaques more commonly used in research, namely the rhesus (M. mulatta) and cynomolgus (M. fascicularis) macaques which are in the fascicularis group. The primary habitat of M. nemestrina is the tropical rainforest, with preferred temperatures of 64° – 86° F (18° – 30° C). Unlike some other macaque species, they do not adapt well to cold environments. Macaques that live in temperate climates tend to exhibit seasonality in breeding to ensure that infants are born in the spring. In contrast, M. nemestrina breed and deliver infants year- round. Young infants are at high risk for developing hypothermia; therefore M. nemestrina colonies should not be maintained outdoors in temperate climates. Although the lack of seasonal breeding raises challenges in maintaining M. nemestrina breeding colonies, it increases their utility as a model for biomedical research.

Infectious Disease Models M. nemestrina demonstrate a high susceptibility to human infectious disease pathogens as well as nonhuman primate viruses, such as simian immunodeficiency virus (SIV) and simian-human chimeric immunodeficiency virus (SHIV).10,14, 35-45 They are of particular interest in HIV/AIDS research due to the presence of a nonfunctional TRIM5α variant (TRIM-cyp) that eliminates a major barrier for replication of HIV-1 in macaques and permits infection with lentivirus strains closely related to HIV-1.46-50 M. nemestrina is the only macaque species to be successfully infected with HIV-151 and are both more easily infected and more likely to exhibit pathologic changes following infection than other macaque species.52 Comparison of pathogenicity in different NHP species provides valuable information on host factors involved in resistance or susceptibility to infection and disease.4,53-57 Mane-A1*004g Mane-A1*006g Macaca nemestrina are also valuable Mane-A3*13g models for non-AIDS infectious disease Mane-A1*010g research, including study of influenza, Mane-A1*018g Mane-A7*01g Mane-A1*019g 58-61 Mane-A6*01g chlamydia, and tuberculosis. One Mane-A1*031g tool that is useful in studying infectious Mane-A4*14g Mane-A1*032g Mane-A1*038g disease is genetic characterization of Mane-A4*13g individual animals, including MHC Mane-A4*01g Mane-A1*040g Mane-A1*063g Mane-A1*052g haplotypes which can affect the immune response. MHC Class I major and Mane-A1*082g minor expressed alleles have been Mane-A4*01/14g analyzed for ~600 animals using the Mane-A1*084g 62 next generation sequencing approach. Mane-A1*108g A representation of the most common Mane-A1*131g Mane-A1*114g MHC I lineages within the M. nemestrina Mane-A2*05g population at the Washington National Primate Research Center is shown here.

Reproductive and Developmental Models M. nemestrina have a less keratinized vaginal epithelium63 and more appropriate vaginal size than M. mulatta, as well as a year-round menstrual cycle. The vaginal flora of M. nemestrina is similar to that of humans.64 These factors make them more useful than other macaque species for evaluating vaginal exposure to pathogens, microbicides and the vaginal microbiome.19,20,22,26,27,39,63,65-71 The results from these microbicide research projects can be directly translated to clinical applications. M. nemestrina exhibit a 32-day menstrual cycle similar to humans, and a gestation length averaging 172 days. Menstrual cycles are easy to track in M. nemestrina; not only do they exhibit obvious menstrual bleeding, but they also display marked ovulatory tumescence, as shown here. This ease of tracking is beneficial for studies which require timed-mating as well as experiments where it is necessary to know the stage of the mestrual cycle for treatment or sample collection. Significant contributions to the understanding of premature delivery and infections during pregnancy have been achieved using the M. nemestrina model.72-78 The fact that M. nemestrina breed year round is advantageous for investigators who perform intensive interventions with pregnant animals or neonates, as research subjects are available year round and experiments can be staggered appropriately. There was a quick response and publication to the Zika virus outbreak that was able to take advantage of the readily available pregnant M. nemestrina to evaluate the effects on the developing primate fetus.79 In addition, treatments for neonatal hypoxia which are now in clinical use were developed using the M. nemestrina model.80-87 M. nemestrina adapt well to a jacket and tether system, and with surgical implantation of catheters and monitors it is possible to administer medication, monitor physiological parameters, and collect biological samples without disturbing the animal. Other Models M. nemestrina are suitable animal models for essentially any project where other macaque species are used. They have been used extensively in research involving hematopoietic stem cell transplantation.13,15,88-107 These projects address a wide range of research topics from the basic biology of immunological tolerance and techniques of gene editing to translational topics of improving transplant success in humans and prevention and elimination of HIV infection. M. nemestrina have also been used to demonstrate functional viability of stem cell transplants in a model of myocardial infarction.108-110 In addition, M. nemestrina models have made significant contributions to neuroscience research.111-131

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