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Erythrocytes of the Rhesus and Cynomolgus Monkeys addresses the morphologic, quantitative, and generative aspects of the erythrocytes of the rhesus monkey Macaca mulatta and the cynomolgus monkey Macaca fascicularis (long-tailed macaque, crab-eating monkey). These two species are the most commonly selected nonhuman primates for basic science and cl
The Rhesus Monkey discusses the use of the rhesus monkey as a test subject in biomedical research. This text provides information that is essential for research that involves the rhesus monkey, from conditioning a wild rhesus monkey to restoring it to its natural state after the tests that it is subjected to. This book covers the phases a rhesus monkey specimen will go through in an experiment. Each chapter covers different facets of using a rhesus monkey as a test subject, including selecting a suitable specimen, conditioning, medication, nourishment, breeding, and pathological threats. Researchers in all fields will find this book an invaluable source of information regarding the best use of rhesus monkeys as research subjects.
Genetic variation in immune genes is known to impact infectious disease progression and outcomes. Rhesus and cynomolgus macaque monkeys are particularly favored as animal models for human diseases because their anatomy, physiology, and immunology closely mimic that of humans. Our reliance on macaque models to understand human disease progression requires a close examination of the functional impacts of their immune gene variation. In this dissertation I focus on two immune loci, the Fc gamma receptors (FCGR) and major histocompatibility complex (MHC), that have been consistently implicated in affecting HIV disease outcomes and vaccine efficacy. In chapter 1, I review what is known about how these genes impact HIV disease outcomes and vaccine efficacy in humans, and what is known about their variation and their impact on simian immunodeficiency virus (SIV) infection in macaques. In chapter 2, I describe our work characterizing FCGR genetic variation and its functional consequences in Mauritian cynomolgus macaques (MCM). These animals have limited genetic diversity and have relatively simple MHC genetics compared to rhesus macaques, making them an attractive choice for simian immunodeficiency virus (SIV) studies. Fc gamma receptors mediate the effects of antibodies, including the broadly neutralizing antibodies that are the holy grail of HIV vaccination. We characterized the genetic diversity of the FCGR genes using long-read sequencing and assessed whether the genetic diversity impacted the ability of variants to bind IgG antibodies. We found that FCGR variation in MCMs does not have a substantial effect on IgG binding. This suggests that researchers using MCMs for studies where antibody responses are a critical outcome may not need to account for FCGR genotype in their animals. In chapter 3, I describe our work using an ultradense peptide array as a high-throughput method for quantifying class I MHC-peptide binding. The class I major histocompatibility complex is responsible for presenting intracellular peptides, including peptides derived from viruses, to CD8 T cells, which mount a cytolytic response against infected cells. Tracking specific CD8 T cell responses requires knowledge of CD8 T cell epitopes - specific peptides that can bind and be presented by MHC molecules and elicit CD8 T cell responses. Screening peptides for binding to MHC class I molecules streamlines identification of these epitopes, and our peptide array approach in-creases by several orders of magnitude the number of peptides that can be tested for MHC binding in a single experiment. We found that for SIV peptides that had been previously found to bind to specific rhesus macaque MHC molecules, the peptide array binding scores were significantly higher than for peptides that had been found to not bind MHC molecules, suggesting that the array can be effectively used as a high-throughput tool for identifying MHC-binding peptides. In chapter 4, I describe our use of the peptide binding datasets generated for 16 rhesus macaque MHC molecules to generate MHC binding motif and peptide binding predictions. We found that, though imperfect, neural net models trained on peptide array data can be used to identify putative CD8 T cell epitopes, including immunodominant epitopes. Taken together, this work expands researchers' ability to effectively use nonhuman primates as models for infectious diseases, and we placed an emphasis on using newer and novel technologies to improve the accuracy and efficiency of our analyses. Our work characterizing FCGR variation and functional impacts in Mauritian cynomolgus macaques enables researchers to more efficiently use these animals for studies involving antibody responses. Our work using ultradense peptide arrays to screen peptides for MHC binding and generating datasets that can be used to predict peptide binding offers an alternative, high-throughput approach to streamline identification of CD8 T cell epitopes. This work lays the groundwork for characterizing less well-studied macaque MHC molecules and rapidly identifying their peptide binding repertoires in large and understudied pathogens.