Macaca Fuscata Research Articles

Diagonal-couplet gaits on discontinuous supports in Japanese macaques and implications for the adaptive significance of the diagonal-sequence, diagonal-couplet gait of primates.

Diagonal-sequence, diagonal-couplet (DSDC) gaits have been proposed as an adaptation to travel on discontinuously arranged arboreal branches. Only a few studies have examined primate gait adjustment to support discontinuity. We analyzed the gaits of Japanese macaques walking on the "ground" and two discontinuous conditions, "circle" and "point," to better understand the advantages of DSDC gaits on discontinuous supports. Seventy-eight vertical posts, each with a circular upper surface, were arranged in four rows at a spacing of 200 mm. The diameter of the circular upper surface was 150 mm ("circle condition") or 50 mm ("point condition"). We calculated the limb phase, duty factor, and time interval from hindlimb touchdown to ipsilateral forelimb liftoff. The supports the fore- and hindlimbs landed on during walking were identified in the circle and point condition. The macaques predominantly used DSDC gaits in the ground and circle conditions and lateral-sequence, diagonal-couplet (LSDC) gaits in the point condition. The macaques usually placed their hindlimbs on the same supports as their ipsilateral forelimbs during the gait cycle. Japanese macaques overlapped the ipsilateral fore- and hindlimb stance phase in all DSDC and some LSDC gaits to proximate the ipsilateral limbs on the discontinuous support, allowing the forelimb to guide the hindlimb placement to the support. The overlap duration of the ipsilateral limb stance phases may be extended by DSDC gaits longer than by LSDC gaits, allowing for a direct pass of the support being held by the prehensile hand to the prehensile foot.

Mothers of disabled infants had higher cortisol levels in a free-ranging group of Japanese macaques (Macacafuscata).

Glucocorticoids (GCs) are hormones released in response to stressors and can provide insight into an organism's physiological well-being. Experiencing chronic challenges to homeostasis is associated with significant deviations from baseline fecal GCs (fGCs) in many species, providing a noninvasive biomarker for assessing stress. In the group of free-ranging Japanese macaques (Macaca fuscata) at the Awajishima Monkey Center in Japan, ~17% have congenital limb malformations. We collected 646 fecal samples from 27 females over three consecutive birth seasons (May-August)and analyzed them using enzyme immunoassay to extract fGCs. We explored the relationship between fGC levels and individual (physical impairment and reproductive status), social (dominance rank and availability of kin for social support), and ecological variables (exposure to potential predators, rainfall, and wild fruit availability). A disabled infant was associated significantly with higher fGC in the mother; however, physical impairment in adult females was not significantly related to fGC levels. Females with higher dominance rank had significantly lower fGC levels than lower ranking females. Other factors did not relate significantly to fGC. These results suggest that providing care that meets the support needs of disabled infants poses a physiological challenge for mothersand suggests that physically impaired adults are able to effectively compensate for their disabilities with behavioral plasticity. Once an individual with congenital limb malformations survives infancy through their mother's care, physical impairment does not appear to influence fGC values, while social variables like dominance rank significantly influenced cortisol values in free-ranging female Japanese macaques.

Effect of Social Dominance and Reproductive State on Adrenal Steroids in Female Japanese Macaques (Macaca fuscata)

Effect of Social Dominance and Reproductive State on Adrenal Steroids in Female Japanese Macaques (Macaca fuscata)

Measuring short-term changes in stress-associated salivary analytes in free-ranging Japanese macaques (Macaca fuscata).

Monitoring short-term changes of endocrine responses in non-human primates living in wild populations is a challenge. Saliva contains enzymes, steroids, and various analytes that can be broadly useful for helping us understand physiological responses to social and environmental sources of stress as well as other compromises to an individual's health homeostasis. Salivary alpha-amylase and salivary cortisol are known to respond rapidly to stress, which can allow us to use these analytes to monitor stress on a time scale of minutes. We developed a non-invasive methodology for the collection of saliva and verified its applicability by conducting short-term interval sampling of focal individuals under varying social conditions in a group of semi-provisioned free-ranging Japanese macaques (Macaca fuscata) living endemically on the island of Koshima, Miyazaki prefecture, Japan. Monkeys were habituated to sampling by using ingestive attractants applied to cotton ropes. Their receptivity to chewing on the saliva sampling material provided and chewing time was recorded ad libitum during the initial habituation period of the troop to the sampling procedure. Focal animal behavior observations were conducted in conjunction with saliva sampling. Salivary analytes associated with stress, alpha-amylase and cortisol, were measured via enzyme immunoassay. Short-term changes in salivary alpha-amylase and salivary cortisol were examined in relation to short-term changes in social behavior, in particular aggression or grooming. We tested flow-rate effects in both analytes and found strong correlations between original sample results and their respective flow-rate transformed equivalents. Additionally, temperature effects on samples were tested and both analytes showed nearly the same values when stored at -20, 4, and 30°C conditions for 6h; important information expected to facilitate future sampling in field conditions where freezer storage is uncertain. Our study shows that saliva can be repeatedly sampled non-invasively to investigate short-term changes in stress-associated markers in free-ranging Japanese macaques.

Mineral acquisition of Japanese macaques: Contents in the foods, digestibility, and sodium-provisioning experiment.

Minerals provide micronutrients that function in various ways in the body, and they are necessary for the survival of animals. In this study, we first compared the mineral content of foods of wild Japanese macaques in lowland Yakushima with that of monkey chow used for many years to feed captive macaques and specifically formulated to obtain good health in captive macaques (National Research Council [NRC] recommendations). Second, we clarified the mineral balance in captive individuals when feeding them monkey chow to clarify the digestibility/bioavailability of the minerals. Third, we investigated the physiological response when we experimentally increased sodium intake. In the lowland of Yakushima, which is in the vicinity (<800 m) of the coast, animals, fungi, and mature leaves had high sodium contents compared with NRC recommendations. The calcium contents of mature leaves and animals were higher than the NRC recommendations. The overall mineral intake in this population was lower than that in the captive animals for calcium, phosphorus, sodium, and iron, while similar for magnesium and higher in potassium. Patterns in the intake and excretion of minerals indicated that excretion was mostly from urine and not from feces, and apparent digestibility was high for sodium. This tendency was opposite for calcium, magnesium, and phosphorus and intermediate for potassium. A sodium-provisioning experiment showed that fecal aldosterone concentration remained low in both control and sodium-provisioning conditions so the macaques do not need reabsorb sodium in the kidneys. Therefore, sodium content in the monkey chow, which is slightly lower than the NRC recommendation, seemed high enough so that the macaques could avoid the need to reabsorb sodium in the kidneys. We advocate similar studies for other primate populations or species to better understand the role of mineral concentrations on food selection and to identify potential mineral deficiencies.

Maternal Western-style diet in nonhuman primates leads to offspring islet adaptations including altered gene expression and insulin hypersecretion.

Maternal overnutrition is associated with increased susceptibility to type 2 diabetes in the offspring. Rodent models have shown that maternal overnutrition influences islet function in offspring. To determine whether maternal Western-style diet (WSD) alters prejuvenile islet function in a model that approximates that of human offspring, we utilized a well-characterized Japanese macaque model. We compared islet function from offspring exposed to WSD throughout pregnancy and lactation and weaned to WSD (WSD/WSD) compared with islets from offspring exposed only to postweaning WSD (CD/WSD) at 1 yr of age. WSD/WSD offspring islets showed increased basal insulin secretion and an exaggerated increase in glucose-stimulated insulin secretion, as assessed by dynamic ex vivo perifusion assays, relative to CD/WSD-exposed offspring. We probed potential mechanisms underlying insulin hypersecretion using transmission electron microscopy to evaluate β-cell ultrastructure, qRT-PCR to quantify candidate gene expression, and Seahorse assay to assess mitochondrial function. Insulin granule density, mitochondrial density, and mitochondrial DNA ratio were similar between groups. However, islets from WSD/WSD male and female offspring had increased expression of transcripts known to facilitate stimulus-secretion coupling and changes in the expression of cell stress genes. Seahorse assay revealed increased spare respiratory capacity in islets from WSD/WSD male offspring. Overall, these results show that maternal WSD feeding confers changes to genes governing insulin secretory coupling and results in insulin hypersecretion as early as the postweaning period. The results suggest a maternal diet leads to early adaptation and developmental programming in offspring islet genes that may underlie future β-cell dysfunction.NEW & NOTEWORTHY Programed adaptations in islets in response to maternal WSD exposure may alter β-cell response to metabolic stress in offspring. We show that islets from maternal WSD-exposed offspring hypersecrete insulin, possibly due to increased components of stimulus-secretion coupling. These findings suggest that islet hyperfunction is programed by maternal diet, and changes can be detected as early as the postweaning period in nonhuman primate offspring.

Influence of radiation exposure to delayed fetal growth in wild Japanese monkeys after the Fukushima accident.

Wild Japanese monkeys (Macaca fuscata) were exposed to radiation after the Fukushima Daiichi nuclear accident in 2011. To clarify the biological effects of radiation exposure on their fetal growth, pregnant monkeys and their fetuses were analyzed. These animals were collected between 2008 and 2020 (before and after the accident in 2011) living in Fukushima City, approximately 70 km from the nuclear power plant. Multiple regression analyses were conducted with fetal body weight (FBW) and fetal head circumference (FHS) as objective variables, and maternal and fetal factors as explanatory variables. The maternal factors were relative exposure dose rate (REDR), age, body weight, body length, fat index, and parity. The fetal factors were crown ramp length (CRL) and sex. Multiple regression analyses showed that FBR and FHS growth were positively associated with CRL, maternal body length, and negatively associated with REDR. Since the relative growth of FBR and FHS to CRL decreased with increasing REDR, radiation exposure due to the nuclear accident may have contributed to the delayed fetal growth observed in Japanese monkeys.

Prosociality in a despotic society

Prosociality is the intent to improve others' well-being. Existing hypotheses postulate that enhanced social tolerance and inter-individual dependence may facilitate prosocial preferences, which may favor the evolution of altruism. While most studies are restricted to "tolerant" (cooperatively breeding and self-domesticated) species, despotic societies provide an alternative opportunity to investigate prosociality due to nepotism and ample inter-individual dependencies. Japanese macaques live in hierarchical matrilineal societies, with strong kin bonds. Besides, tolerance among non-kin may persist through reciprocity. Using a group service food-provision paradigm, we found prosocial preferences in a semi free-ranging group of Japanese macaques. The extent of provisioning was at levels comparable to tolerant species. Dyadic tolerance predicted the likelihood and magnitude of provisioning, while kinship predicted the magnitude. We emphasize the role of a complex socio-ecology fostering individual prosocial tendencies through kinship and tolerance. These findings necessitate a framework including different forms of interdependence beyond the generally tolerant species.

Behavioral responses of solicitors after failure to receive grooming in Macaca fuscata

Grooming is one of the most common cooperative behaviors among several animal species. However, the tactics used to cope with uncooperative partners in grooming interactions remain unclear. Japanese macaques (Macaca fuscata) solicit grooming from partners through postural behaviors, but may not necessarily receive grooming. This study investigated the behavior of female Japanese macaques after they solicited but did not receive grooming. We predicted that unsuccessful solicitors would engage in grooming interactions with uncooperative partners if they were affiliated. If they were not affiliated, the solicitors would not do so and may seek grooming interactions with other grooming partners. We used a focal-animal sampling method, targeting 17 females at Katsuyama, Okayama Prefecture, Japan. We recognized affiliative relationships by measuring close spatial association. After unsuccessful solicitation, females tended to scratch themselves, suggesting that solicitors may experience anxiety or distress when they do not receive grooming. They also tended to be proximate with affiliated partners after solicitation, regardless of whether the solicitors received grooming from their partners. In contrast, when solicitors failed to receive grooming from unaffiliated partners, their subsequent proximity was lower than when they were groomed. Moreover, unsuccessful solicitors were likely to engage in grooming interactions with affiliated partners who were uncooperative (receivers of unsuccessful solicitations). However, they were less likely to engage in grooming interactions with unaffiliated partners and instead engaged in grooming interactions with other nearby partners. These findings indicate that female Japanese macaques decide whether to engage in grooming interactions with uncooperative partners who have not groomed solicitors based on affiliative relationships and the availability of other grooming partners. It is probable that, when the cost of searching for a grooming partner is low, female Japanese macaques are likely to switch partners, potentially leading to an increase in the benefits obtained from grooming interactions.

Coordination within paraspinal muscles during bipedalism in humans, a white-handed gibbon, and a Japanese macaque

Coordination within paraspinal muscles during bipedalism in humans, a white-handed gibbon, and a Japanese macaque

Emergence of an erythroid cell-specific regulatory region in ABO intron 1 attributable to A- or B-antigen expression on erythrocytes in Hominoidea

A- and B-antigens are present on red blood cells (RBCs) as well as other cells and secretions in Hominoidea including humans and apes such as chimpanzees and gibbons, whereas expression of these antigens on RBCs is subtle in monkeys such as Japanese macaques. Previous studies have indicated that H-antigen expression has not completely developed on RBCs in monkeys. Such antigen expression requires the presence of H-antigen and A- or B-transferase expression in cells of erythroid lineage, although whether or not ABO gene regulation is associated with the difference of A- or B-antigen expression between Hominoidea and monkeys has not been examined. Since it has been suggested that ABO expression on human erythrocytes is dependent upon an erythroid cell-specific regulatory region or the + 5.8-kb site in intron 1, we compared the sequences of ABO intron 1 among non-human primates, and demonstrated the presence of sites orthologous to the + 5.8-kb site in chimpanzees and gibbons, and their absence in Japanese macaques. In addition, luciferase assays revealed that the former orthologues enhanced promoter activity, whereas the corresponding site in the latter did not. These results suggested that the A- or B-antigens on RBCs might be ascribed to emergence of the + 5.8-kb site or the corresponding regions in ABO through genetic evolution.

Diurnal variation in declarative memory and the involvement of SCOP in cognitive functions in nonhuman primates

Cognitive functions depend on the time of day in various organisms. Previously, we found that 24-h recognition memory performance of nocturnal mice changes diurnally through SCOP protein-dependent regulation. It remains unknown whether diurnal change and SCOP-dependent regulation of memory performance are conserved across species with diurnal/nocturnal habits. We tested whether the memory performance of diurnal Japanese macaques depends on the time of day. The memory association between bitter taste of drinking water and the nozzle color of the water bottle was established during the light period of the day to evaluate of memory performance for macaques. Here we found diurnal variation of declarative memory in Japanese macaques. The middle of the daytime is the most effective time for memory performance during the light period. To assess whether SCOP is involved in declarative memory performance, we interfered with SCOP expression by using lentiviral vector expressing shRNA against Scop in the hippocampus of Japanese macaques. Scop knockdown in the hippocampus abrogated the memory performance in the middle of the daytime. Our results implicate that SCOP in the hippocampus is necessary for the diurnal rhythm of the memory system and that the SCOP-dependent memory regulation system may be conserved in mammals.

Dehydroepiandrosterone sulfate (DHEAS) in excreta is a good indicator of serum DHEAS in Japanese macaques (Macaca fuscata)

We developed a microplate enzyme immunoassay (EIA) to measure dehydroepiandrosterone sulfate (DHEAS) in the blood, urine, and feces of Japanese macaques and evaluated the relationship between serum DHEAS and excreta DHEAS. Our DHEAS EIA using heterological DHEA derivatives conjugated with enzyme was highly sensitive, and linearities and recoveries for all matrices of Japanese macaques were reliable. For the biological evaluation of the metabolism of DHEAS in Japanese macaques, dissolved DHEAS was injected into the subjects, and consecutively collected serum, urine, and fecal samples were analyzed. The peaks of serum DHEAS were observed 6 h after the administration, while those of urine and feces were observed after 24 h. The fluctuation of those in urine and feces were significantly correlated with serum DHEAS levels. In addition, we measured pregnanediol-glucuronide (PdG), and estrone-conjugate (E1C) in urine and fecal samples to investigate the effects of administrated DHEAS on these progesterone and estrogen metabolites. The peak of PdG was observed 24 h after administration, then declined sharply. The concentration of E1C increased 1 week after administration in two out of three subjects. Our results suggest that measuring urinary and fecal DHEAS with our EIA provides a non-invasive alternative to assessing DHEAS levels in the serum of Japanese macaques.

Muscle-tendon arrangement and intramuscular nerve distribution of flexor digitorum superficialis in the siamang (Symphalangus syndactylus), western lowland gorilla (Gorilla gorilla gorilla), western chimpanzee (Pan troglodytes verus), and Japanese macaque (Macaca fuscata).

Flexor digitorum superficialis (FDS) shows diverse muscle-tendon arrangements among primates. The intramuscular nerve distribution pattern is a criterion for discussing the homology of FDS. In this study, the muscle-tendon arrangement and intramuscular nerve distribution of FDS were examined in the siamang, western lowland gorilla, western chimpanzee, and Japanese macaques. The FDS had muscle bellies to digits II-V. FDS had proximal belly and intermediate tendon except for siamang. Distal belly to digit II (in the western lowland gorilla and western chimpanzees) or distal bellies to digits II and V (in Japanese macaque) originated from the intermediate tendon. In all specimens, nerve branches within digit III belly extended into digit IV belly, and nerve branch(es) within digit IV belly extended into digit V belly. This consistent pattern suggested that each muscle belly to digits III-V is interspecifically homologous. The digit II belly in the siamang and the distal belly to digit II in the western lowland gorilla, western chimpanzees, and Japanese macaques could be homologous based on their similar innervating patterns. The proximal belly was innervated by branches from the communicating nerve between median and ulnar nerves in the western lowland gorilla or branches from median and ulnar nerves in western chimpanzees. In the siamang and Japanese macaque, the whole FDS was innervated by median nerve. The proximal belly in the western lowland gorilla, western chimpanzees, and Japanese macaques could be classified into different groups from the other part of the FDS.

Sex-Specific Effects of Juvenile Offspring on their Mothers’ Social Relationships with Other Females in Japanese Macaques (Macaca fuscata) on Shodoshima Island

Sex-Specific Effects of Juvenile Offspring on their Mothers’ Social Relationships with Other Females in Japanese Macaques (Macaca fuscata) on Shodoshima Island

Emergent centrality in rank-based supplanting process.

We propose a stochastic process of interacting many agents, which is inspired by rank-based supplanting dynamics commonly observed in a group of Japanese macaques. In order to characterize the breaking of permutation symmetry with respect to agents' rank in the stochastic process, we introduce a rank-dependent quantity, overlap centrality, which quantifies how often a given agent overlaps with the other agents. We give a sufficient condition in a wide class of the models such that overlap centrality shows perfect correlation in terms of the agents' rank in the zero-supplanting limit. We also discuss a singularity of the correlation in the case of interaction induced by a Potts energy.

Molecular epidemiology of Bartonella quintana endocarditis in patients from Israel and Eastern Africa

BackgroundBartonella quintana is an important cause of culture-negative endocarditis. Although humans have been considered as its only reservoir, recent studies showed that macaque species are also reservoirs of B. quintana. Based on multi-locus sequence typing (MLST) B. quintana strains have been classified into 22 sequence types (STs), with 7 STs exclusively found in humans. Data regarding the molecular epidemiology of B. quintana endocarditis is limited to only 3 STs identified in 4 patients from Europe and Australia. We studied B. quintana endocarditis acquired in Eastern Africa or Israel to investigate the genetic diversity and clinical relatedness of B. quintana from distinct geographic regions.MethodsEleven patients with B. quintana endocarditis, 6 from Eastern Africa and 5 from Israel, were studied. DNA was extracted from cardiac tissue or blood specimens and analyzed by MLST based on 9 genetic loci. An evolutionary relationship between STs was visualized by a minimum spanning tree. A phylogenetic tree was constructed with the concatenated sequences (4271 bp) of the 9 loci using the maximum-likelihood method.ResultsSix strains were classified into previously described STs while 5 strains were identified for the first time and classified into new STs 23–27 which clustered with the previously reported STs 1–7 from human strains found in Australia, France, Germany, the USA, Russia, and the former Yugoslavia, without indication of geographical structuring. ST2 was the most prevalent ST, found in 5 of 15 patients with endocarditis (33.3%). ST26 appears to be a primary founder of the human lineage.ConclusionsThe new and previously reported human STs form a single human lineage, clearly separated from the other 3 B. quintana lineages of cynomolgus, rhesus, and Japanese macaques. From evolutionary perspectives, these findings support the assumption that B. quintana has co-evolved with host species to form a host-speciation pattern. ST26 is suggested herein as a primary founder of the human lineage and may be key to explore where B. quintana had first originated; ST2 is a dominant genetic type associated with B. quintana endocarditis. To confirm these findings, additional worldwide molecular epidemiological studies are required.

Monkey Damage Management in Japan: A Case Study in Hyogo Prefecture, Western Japan

Agricultural damage caused by wild Japanese monkeys (Macaca fuscata) has become a serious problem in Japan. Residents have tried to protect their crop fields from monkey invasions, but the excellent physical and learning abilities of the monkeys have made the protection difficult. At present, many local governments are attempting to reduce the damage mainly through culling, but unplanned population management often results in over-culling, which consequently leads to local extinction. Here, I introduce a case study in Kami Town, Hyogo Prefecture, western Japan, in which the local government simultaneously implemented three countermeasures: 1) organizing a monkey patrol group, 2) expanding inexpensive and easy-to-use electric fences, and 3) selective capturing of problem animals. After the implementation of these countermeasures, frequency of chasing away the monkeys around crop field (a total of 319 times in 2010 compared to less than 100 times in 2017) and amount of agricultural damage (2011: 2 925 000 JPY, 2017: 876 000 JPY) decreased. These results showed that in order to decrease agricultural damage while maintaining a stable monkey population, a combination of multiple countermeasures is a good solution, and methods applied in Kami Town should also be implemented in other areas where monkeys regularly damage crops.

Genetic effects of demographic bottleneck and recovery in Kinkazan Island and mainland populations of Japanese macaques (Macaca fuscata).

Populations of Japanese macaques were significantly reduced in most areas from the 1900s to the 1960s and then recovered mainly in the northeastern part of Honshu. A drastic reduction in population size reduces genetic variability through a bottleneck effect. Demographic expansion after the reduction that accumulates new mutations can reduce the bottleneck effects or drive the recovery of genetic variability. We examined the genetic status of a small island population (Kinkazan Island) and a larger mainland population (southern Tohoku) of Japanese macaques that experienced recent demographic bottlenecks and recovery using eight microsatellite loci. The two populations were significantly genetically different from each other. The Kinkazan population exhibited lower genetic variability, remarkable evidence of bottleneck (i.e., significant heterozygosity excess and lower frequency of rare alleles), and a considerably smaller effective population size based on genetic data than based on the current census size. These results indicate that the genetic status has not completely recovered from the demographic bottleneck despite a full recovery in census size on Kinkazan Island. New mutations might rarely have accumulated because of the small carrying capacity of the island. Therefore, the genetic variability of the population would have been restrained by the severe bottleneck size, small carrying capacity, and long-term isolation. On the other hand, the bottleneck effect seems to be limited in the southern Tohoku population considering higher genetic variability, non-significant heterozygosity excess in many mutation conditions, and the highest frequency of rare alleles.

Vaccination with short-term-cultured autologous PBMCs efficiently activated STLV-1-specific CTLs in naturally STLV-1-infected Japanese monkeys with impaired CTL responses.

A small proportion of human T-cell leukemia virus type-1 (HTLV-1)-infected individuals develop adult T-cell leukemia/lymphoma, a chemotherapy-resistant lymphoproliferative disease with a poor prognosis. HTLV-1-specific cytotoxic T lymphocytes (CTLs), potential anti-tumor/virus effectors, are impaired in adult T-cell leukemia/lymphoma patients. Here, using Japanese monkeys naturally infected with simian T-cell leukemia/T-lymphotropic virus type-1 (STLV-1) as a model, we demonstrate that short-term-cultured autologous peripheral blood mononuclear cells (PBMCs) can serve as a therapeutic vaccine to activate such CTLs. In a screening test, STLV-1-specific CTL activity was detectable in 8/10 naturally STLV-1-infected monkeys. We conducted a vaccine study in the remaining two monkeys with impaired CTL responses. The short-term-cultured PBMCs of these monkeys spontaneously expressed viral antigens, in a similar way to PBMCs from human HTLV-1 carriers. The first monkey was subcutaneously inoculated with three-day-cultured and mitomycin C (MMC)-treated autologous PBMCs, and then boosted with MMC-treated autologous STLV-1-infected cell line cells. The second monkey was inoculated with autologous PBMC-vaccine alone twice. In addition, a third monkey that originally showed a weak STLV-1-specific CTL response was inoculated with similar autologous PBMC-vaccines. In all three vaccinated monkeys, marked activation of STLV-1-specific CTLs and a mild reduction in the STLV-1 proviral load were observed. Follow-up analyses on the two monkeys vaccinated with PBMCs alone indicated that STLV-1-specific CTL responses peaked at 3-4 months after vaccination, and then diminished but remained detectable for more than one year. The significant reduction in the proviral load and the control of viral expression were associated with CTL activation but also diminished 6 and 12 months after vaccination, respectively, suggesting the requirement for a booster. The vaccine-induced CTLs in these monkeys recognized epitopes in the STLV-1 Tax and/or Envelope proteins, and efficiently killed autologous STLV-1-infected cells in vitro. These findings indicated that the autologous PBMC-based vaccine could induce functional STLV-1-specific CTLs in vivo.